interacts_with DB00783?
The heat shock protein 90 inhibitor 17- P29372 suppresses growth and induces apoptosis in human cholangiocarcinoma cells . The aim of this study was to investigate the effects of 17-Allylamino-17-demethoxygeldanamycin ( 17- P29372 ) , a heat shock protein 90 ( HSP90 ) inhibitor , on the proliferation , cell cycle , and apoptosis of human cholangiocarcinoma ( CCA ) cells . Cell proliferation and cell cycle distribution were measured by the MTT assay and flow cytometry analysis , respectively . Induction of apoptosis was determined by flow cytometry and Hoechst staining . The expressions of cleaved poly ADP-ribose polymerase ( PARP ) , Bcl-2 , Survivin , and P12004 B1 were detected by Western blot analysis . The activity of caspase-3 was also examined . We found that 17- P29372 inhibited cell growth and induced G2/M cell cycle arrest and apoptosis in CCA cells together with the down-regulation of Bcl-2 , Survivin and P12004 B1 , and the up-regulation of cleaved PARP . Moreover , increased caspase-3 activity was also observed in CCA cells treated with 17- P29372 . In conclusion , our data suggest that the inhibition of HSP90 function by 17- P29372 may provide a promising therapeutic strategy for the treatment of human CCA . P00747 activator and plasminogen activator inhibitor gene expression in human saphenous vein organ culture . We describe the expression of tissue-type plasminogen activator ( t-PA ) and plasminogen activator inhibitor ( P05121 ) in saphenous vein culture . Smooth muscle cells ( SMC ) are quiescent in fresh tissue , whereas these cells acquire a proliferating phenotype when venous segments are cultured in the presence of serum . t-PA and P05121 were localized immunohistochemically and quantified using biochemical techniques . t-PA and P05121 mRNA was quantified by reverse transcription polymerase chain reaction ( RT -- PCR ) assay . Immunostaining showed an increase in the positivity of proliferating cell nuclear antigen ( P12004 ) from 10-day tissue culture . Tissue sections from fresh vein showed minimal t-PA and maximal P05121 immunostaining . In contrast , 10-day cultures showed an increase in t-PA and a decline in P05121 staining . Biochemical analysis revealed a 118 % increase in t-PA and a 50 % decrease in P05121 antigen levels from 10-day tissue cultures . RT -- PCR demonstrated that the mRNA encoding t-PA increased , while P05121 decreased after 10 days of culture . In conclusion , venous culture showed an up-regulation of t-PA and a repression of P05121 gene expression during SMC proliferation in the vessel wall . The P05121 repression observed in venous culture is in contrast to the situation observed in human atheroma . A shift in the t-PA/ P05121 balance may have a role in vascular remodeling . [ Effects of octreotide on necrosis of hepatocellular carcinoma xenografts in nude mice ] . BACKGROUND AND OBJECTIVE : DB00104 , a kind of somatostatin analogue , may inhibit the growth of hepatocellular carcinoma ( HCC ) . This study was to investigate the mechanism of inducing necrosis of HCC xenografts in nude mice by octreotide . METHODS : The proliferation of HepG2 cells was determined by MTT assay . Nude mice bearing HepG2 xenografts were treated with octreotide [ 100 microg times ; ( kg times ; d ) (-1) ] or normal saline ( as control ) for eight weeks . The necrosis of HCC was estimated by histology . Vascular endothelial growth factor ( P15692 ) was detected by immunohistochemistry . Somatostatin receptor 2 ( P30874 ) was quantified by Western blot and located with immunohistochemistry . RESULTS : The proliferation of HepG2 cells was not obviously affected by 24-hour treatment of octreotide ( 0.1-1000 nmol/L ) in vitro . The tumor weight was significantly heavier in octreotide group than in control group [ ( 7.15+/-2.96 ) g vs. ( 4.21+/-3.11 ) g , P < 0.05 ] , while the proportion of necrotic volume was significantly higher in octreotide group than in control group [ ( 81.86+/-0.05 ) % vs. ( 43.75+/-0.06 ) % ,P < 0.05 ] . In contrast with control group , P15692 was undetected in the xenografts in octreotide group . P30874 expression in xenograft sinusoids was similar in both groups . CONCLUSION : With active proliferation of HCC cells , octreotide can induce necrosis in HCC xenografts only through the inhibition of angiogenesis mediated by P30874 in the tumor . Engineering the response to vascular injury : divergent effects of deregulated Q01094 expression on vascular smooth muscle cells and endothelial cells result in endothelial recovery and inhibition of neointimal growth . P01375 -alpha ( P01375 ) is expressed locally in the vessel wall after angioplasty and induces growth arrest and apoptosis in endothelial cells ( ECs ) , thereby delaying reendothelialization . Prior studies have shown that direct antagonism of P01375 , using a systemically administered soluble receptor , can enhance endothelial recovery and reduce neointimal thickening . These studies have also shown that downregulation of the transcription factor Q01094 was a key mechanism of P01375 's effect on ECs . We now show that Ad- Q01094 overexpression at sites of balloon injury accelerates functional endothelial recovery , consistent with the prior in vitro findings . Moreover these studies also reveal divergent effects of P01375 and overexpression of Q01094 on ECs versus VSMCs . P01375 exposure of VSMCs had no affect on proliferation or apoptosis , in contrast to the effect seen in ECs . In Ad- Q01094 -transduced VSMCs , however , P01375 -induced marked apoptosis in contrast to the survival effect seen in ECs . Finally , these studies suggest that differential activation of NF-kappaB may play a key role in mediating these opposing effects . Nuclear translocation and transcriptional activity of NF-kappaB was markedly attenuated in Ad- Q01094 -transduced VSMCs , whereas it remained active in similarly treated ECs when the cells were exposed to P01375 . These studies reveal that overexpression of Ad- Q01094 primes VSMCs to P01375 -induced apoptosis . Furthermore , Q01094 potentiates VSMC death by blocking antiapoptotic signaling pathway through inhibition of NF-kappaB activation . The divergent responses of VSMCs and ECs to Q01094 overexpression provide unique therapeutic possibilities : simultaneously targeting the cell cycle of two different cell types , within same tissue microenvironment resulting in opposite and biologically complimentary effects . Synthetic delivery system for tuberculosis vaccines : immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable P00747 microparticles . Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines , particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable . The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly ( DL-lactide co-glycolide ) ( P00747 ) microparticles which served as a delivery system . Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund 's adjuvant ( IFA ) . Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund 's adjuvant ( IFA ) . T-cell responses were found to be superior to those induced with antigen/IFA . The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA . Differences in adjuvanticity were revealed by measuring antigen-specific IgG1 , IgG2a and antigen-induced P01579 secretion in vitro : substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA . This was paralleled by a tenfold higher secretion of P01579 in mice injected with antigen/microparticles . Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli . Taken together these results indicate that biodegradable P00747 microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis . In vitro analysis of the E1A-homologous sequences of Q13029 . The Q13029 ( G3B/MTB-Zf ) gene was first isolated based on its ability to bind to the retinoblastoma protein ( P06400 ) . An acidic , approximately 100-amino-acid region around the P06400 -binding motif of Q13029 has structural and antigenic similarity to the conserved sequences of the E1A viral oncogene . We show here that this region interacts specifically with the E1A-binding domain of P06400 . This interaction could be disrupted by E1A or by a peptide of Q13029 homologous to the P20023 motif of E1A which is involved in binding to P06400 family proteins . Also like E1A , Q13029 can form a ternary complex with P06400 and Q01094 . Despite this similarity to E1A , however , Q13029 could not bind to the P06400 family proteins P28749 and Q08999 in vitro . The data show that the Q13029 P20023 motif can mediate differential binding to P06400 family proteins . We also mapped the shared antigenic determinant between Q13029 and E1A to a conserved sequence , designated CE1 , which is located in the C terminus of E1A . Unlike that of P25101 , the CE1 motif of Q13029 is located next to the P20023 motif . Despite this proximity , CE1 and P20023 appear to act independently . The data show similarities as well as differences between the homologous sequences of Q13029 and E1A and contribute to an understanding of the biochemistry of these proteins . Preferred therapies for neovascular age-related macular degeneration . PURPOSE OF REVIEW : This report reviews the current treatment strategies and the most recent clinical trials in the treatment of neovascular age-related macular degeneration . RECENT FINDINGS : The functional and anatomic outcomes achieved in the pivotal ranibizumab trials with monthly injections set the standard for comparison . Since then , various modified dosing regimens with the aim of lessening the treatment burden associated with monthly injections have been investigated . Additionally , level I evidence now exists for the noninferiority of bevacizumab , as compared to ranibizumab , in the treatment of neovascular age-related macular degeneration ( AMD ) through 1 year of follow-up . DB08885 has emerged as a new anti- vascular endothelial growth factor ( P15692 ) therapy showing encouraging treatment results at 1 year . Novel treatments combined with anti- P15692 agents such as localized radiation are currently being investigated . SUMMARY : Anti- P15692 monotherapy remains the preferred therapy for the management of neovascular AMD at the present time . DB08885 is a new , FDA-approved , effective , anti- P15692 agent available for clinical use . Ongoing clinical trials will help determine the optimal dosing regimens for all of these agents , as well as the long-term efficacy and safety of combination therapy modalities . Point mutation of the somatostatin receptor 2 gene in the human small cell lung cancer cell line COR-L103 . The effect of somatostatin ( SS ) on adrenocorticotrophic hormone ( DB01285 ) secretion from COR-L103 cells derived from a human small cell lung carcinoma was examined . SS at 1 microM had no effect on DB01285 secretion from the cells on either short-term or long-term incubation . Studies by the reverse transcription-polymerase chain reaction ( RT-PCR ) showed that mRNA transcripts of the somatostatin receptor ( SSTR ) 2 , P32745 and P31391 genes were present in COR-L103 cells . Extra bands were obtained by PCR-single strand conformation polymorphism ( SSCP ) analysis of the P30874 gene Sequence analysis of the P30874 gene demonstrated one point mutation in codon 188 of TGG for tryptophan to TGA for a stop codon causing loss of 182 C-terminal amino acid residues of P30874 . The nucleotide sequences of the P32745 and P31391 genes in COR-L103 cells were normal . Binding studies using 125I-Tyr11-SS-14 showed specific affinity binding sites on COR-L103 cells and mouse pituitary tumor AtT-20 cells . DB00104 acetate suppressed the binding of 125I-Tyr11-SS-14 to these two cell lines , but the Kd of COR-L103 cells ( 160 nM ) was 60-fold higher than that of AtT-20 cells ( 2.6 nM ) . Affinity cross-linking studies using 125I-Tyr11-SS-14 gave three bands of 72 KDa , 55 KDa and 32 KDa from AtT-20 cells , but only two bands of 55KDa and 32kDa from COR-L103 cells . These findings suggest that P30874 is not expressed in the plasma membranes of COR-L103 cells due to a point mutation , but that this may have no influence on the effect of SS on DB01285 secretion . Distribution patterns of P12004 and P01160 in perinatal stages of the developing rat heart . Distribution patterns of proliferating cell nuclear antigen ( P12004 ) and atrial natriuretic peptide ( P01160 ) were determined immunohistochemically and morphometrically in atrium and ventriculum of the developing rat heart in different stages of the perinatal period . During the prenatal period , P12004 and P01160 were localized in opposite patterns , particularly in trabecular myocytes . A distinct reduction in the percentage of P12004 -positive nuclei was detected starting at day 19 of the prenatal period , and these cells were rarely observed on postnatal days 30 and 60 . In cardiomyocytes , a distinct increase in P01160 positivity was found , whereas P12004 positivity was very low . It is concluded that P12004 expression gradually decreased from prenatal day 19 onwards , whereas P01160 expression increased in atria throughout the prenatal and postnatal periods , except for a decrease in P01160 expression in ventricles from prenatal day 21 onwards . The opposite expression patterns of P12004 and P01160 in trabecular myocytes of ventricles indicate that P01160 may have antimitogenic/antiproliferative effects in trabecular myocytes . Blocking both type A and B endothelin receptors in the kidney attenuates renal injury and prolongs survival in rats with remnant kidney . Renal disease progression in the rat is associated with a time-dependent upregulation of renal endothelin-1 ( ET-1 ) gene expression and synthesis . We have previously demonstrated that endothelin A receptor subtype ( P25101 ) blockade in rats with remnant kidney reduced signs of disease activity , suggesting that ET-1 exerts part of its deleterious effects on the kidney through P25101 . No data are available so far on the role of ETB receptor in progressive renal injury . We first studied renal P25101 and ETB receptor gene expression in rats with remnant kidney on days 7 , 30 , and 120 after the surgical procedure . While renal expression of P25101 was unaffected , ETB receptor gene was significantly upregulated with time in rats with remnant kidney , being 3.5-fold and sixfold higher than shamoperated rats at days 30 and 120 . We also evaluated whether DB00559 , a nonpeptidic P25101 and ETB receptor antagonist , offered better protection against renal disease progression than reported for P25101 -selective blockers and whether it improved survival in animals with renal ablation . Two groups of rats with renal mass reduction ( n = 11 each ) were given DB00559 100 mg/kg/d orally or its vehicle ( carboxymethyl cellulose ) beginning day 7 after the surgical procedure and were followed until the death of the vehicle-treated animals . Sham-operated animals comprised the control group . DB00559 partially prevented increases in blood pressure and proteinuria , but had a remarkable protective effect on renal function and significantly prolonged animal survival . These data suggest that blocking both renal P25101 and ETB receptors might have major implications in the treatment of human progressive nephropathies . DB08932 ( Opsumit ) for the treatment of pulmonary arterial hypertension . The endothelin pathway is a key pathway for the pathogenesis of pulmonary arterial hypertension ( PAH ) . Antagonism of this pathway is recommended as initial therapy in low-risk patient with PAH to inhibit fibrosis , cell proliferation , and inflammation caused by endothelin . Prior to October 2013 , ambrisentan , a selective P25101 receptor antagonist and DB00559 , a dual P25101 /ETB antagonist , were the only currently available agents for PAH targeting the endothelin pathway . Based on the results of the SERAPHIN trial , macitentan ( brand name Opsumit® ) , a new P25101 /ETB antagonist , has been US FDA approved to delay disease progression and reduce hospitalizations for PAH . SERAPHIN is the first ERA trial to use an event-driven strategy with a composite primary end point of morbidity or mortality . Previous trials have focused on short-term outcomes , such as improved 6-min walk distance and WHO functional class . P15692 trap as a novel antiangiogenic treatment currently in clinical trials for cancer and eye diseases , and VelociGene- based discovery of the next generation of angiogenesis targets . The concept that tumors can be controlled by directly targeting their vascular supply has finally come of age , because clinical trials using a humanized monoclonal antibody that blocks P15692 have demonstrated exciting efficacy in cancer patients , as well as in vascular eye diseases that can lead to blindness . However , data suggest that these current regimens may not provide complete P15692 inhibition and , thus , that the maximum therapeutic potential of P15692 blockade has not yet been achieved . We describe the status of a very potent and high-affinity P15692 blocker , termed the DB08885 , that may provide the opportunity to maximize the potential of P15692 blockade in cancer as well as in vascular eye diseases . We also describe use of the DB08885 as a research tool , when coupled to high-throughput mouse genetics approaches such as VelociGene that can be exploited in strategies to discover and validate the next generation of angiogenesis targets . Effects of systemic injections of vilazodone , a selective serotonin reuptake inhibitor and serotonin 1A receptor agonist , on anxiety induced by predator stress in rats . We examined the effect of DB06684 , a selective serotonin reuptake inhibitor ( SSRI ) and serotonin 1A ( 5-HT(1A) ) receptor agonist [ Bartoszyk , G.D. , Hegenbart , R. , Ziegler , H. , 1997. P50402 68843 , a serotonin reuptake inhibitor with selective presynaptic P08908 receptor agonistic properties. Eur. J. Pharmacol. 322 , 147-153. ] , on change in affect following predator stress . DB06684 and vehicle injection ( intraperitoneal ) occurred either 10 min after predator stress ( prophylactic testing ) , or 90 min prior to behavioral testing for the effects of predator stress ( therapeutic testing ) . Predator stress involved unprotected exposure of rats to a domestic cat . Behavioral effects of stress were evaluated with hole board , plus-maze , and acoustic startle tests 1 week after stress . Predator stress increased anxiety-like behavior in the plus-maze and elevated response to acoustic startle . In prophylactic testing , DB06684 affected stress potentiation of startle at doses above 5 mg/kg . DB06684 increased stress elevation of startle at 10 mg/kg . Higher doses of DB06684 ( 20 and 40 mg/kg ) blocked stress potentiation of startle . In contrast , DB06684 had no effect on stress potentiation of anxiety in the plus-maze . In therapeutic testing , DB06684 increased stress elevation of startle at all doses . In contrast , therapeutic DB06684 had no effect on stress potentiation of anxiety in the plus-maze . Taken together , the data suggest a prophylactic potential for DB06684 in the treatment of changes in hypervigilance following severe stress . DB11558 inhalation-induced adrenocortical hypertrophy and endocrinological changes in rat . Rats were exposed to toluene ( 1,500 ppm for 4 hr per day ) for 7 days . The body weight of the rats was significantly lower and the weight of the adrenal gland was significantly higher in the toluene inhalation group compared to the controls . Microscopically , there was no obvious change in the medulla , but hypertrophy of the cortex was observed in the toluene inhalation group . And , the size of adrenocortical cells in treated-rats was also significantly enlarged than the control . Immunohistochemical staining did not show a clear difference in localization of aldosterone-positive cells between the control and inhalation groups . Expansion of the corticosterone-positive area consistent with the cortical hypertrophy was recognized in the inhalation group . Enhancement of 72 kD-heat-shock protein ( HSP70 ) -expression in the toluene inhalation group was not observed . Neither stress nor damage to cortical cells due directly to toluene exposure was observed in the cortex . Also , there was no obvious difference in the anti-proliferating cell nucleus antigen ( P12004 ) -immunostaining between control and inhalation groups . Thus , it is suspected that cortical hypertrophy was the result of cell enlargement due to the stimulation of the cortical cells . P06850 ( CRF ) immunoreactivity in the paraventricular nucleus ( PVN ) was increased in the inhalation group . Concentration of plasma DB01285 was elevated significantly by toluene exposure . The amounts of mRNA of adrenocortical steroid metabolism gene , cytochrome side-chain cleavage ( P450scc ) , was also increased by toluene inhalation . DB11558 exposure might induce adrenocortical hypertrophy via the hypothalamus-pituitary-adrenal gland ( Q9Y251 ) axis . Q01094 -mediated transcriptional inhibition of the plasminogen activator inhibitor type 1 gene . Gene expression of the plasminogen activation system is cell-cycle dependent . Previously , we showed that ectopic expression of Q01094 repressed the plasminogen activator inhibitor type 1 ( P05121 ) promoter in a manner dependent on the presence of DNA-binding and transactivation domains of Q01094 but independent of binding to pocket-binding proteins , suggesting a novel mechanism for E2F-mediated negative gene regulation [ Koziczak , M. , Krek , W. & Nagamine , Y. ( 2000 ) Mol. Cell. Biol. 20 , 2014-2022 ] . However , it remains to be seen whether endogenous E2F can exert a similar effect . We report here that down-regulation of P05121 gene expression correlates with an increase in endogenous E2F activity . When cells were treated with a cdk2/4-specific inhibitor , which maintains E2F in an inactive state , the decline of serum-induced P05121 mRNA levels was suppressed . In mutant U2OS cells expressing a temperature-sensitive retinoblastoma protein ( pRB ) , a shift to a permissive temperature induced P05121 mRNA expression . In U2OS cells stably expressing an Q01094 -estrogen receptor chimeric protein that could be activated by tamoxifen , P05121 gene transcription was markedly reduced by tamoxifen even in the presence of cycloheximide . These results all indicate that endogenous E2F can directly repress the P05121 gene . P24855 hypersensitive-site analysis of the P05121 promoter suggested the involvement of conformation changes in chromatin structure of the P05121 promoter . 5' deletion analysis of the P05121 promoter showed that multiple sites were responsible for the E2F negative regulation , some of which were promoter dependent . Interestingly , one of these sites is a p53-binding element . Proinflammatory cytokines downregulate gene expression and activity of constitutive nitric oxide synthase in porcine pulmonary artery endothelial cells . We evaluated the effects of cytokines on the catalytic activity and expression of porcine pulmonary artery endothelial cell ( PAEC ) constitutive ( P29474 ) and inducible ( P35228 ) isoforms of nitric oxide synthase ( NOS ) . Exposure of PAEC to the combination of P01579 , P01375 , and P01584 did not alter P35228 activity in cytosolic and membrane fractions but significantly ( p < 0.01 ) reduced P29474 activity in the membrane fraction , but not in the cytosolic fraction , after a 24-h exposure . The cytokine-induced loss of membrane fraction P29474 activity was associated with significant reductions of P29474 mRNA and protein content ( p < 0.01 for both ) . Treatment with the protein synthesis inhibitor , cycloheximide , but not the transcriptional inhibitor actinomycin D prevented cytokine-induced reduction of P29474 mRNA expression . These results suggest that cytokine-induced loss of catalytic activity of P29474 is associated with a reduction in P29474 mRNA and protein mass and that cytokines alter P29474 mRNA stability . Inhibition of protein synthesis prevented reduction of P29474 mRNA by cytokines , suggesting that the mechanism by which cytokines alter P29474 mRNA stability involves protein synthesis . Compensatory increases in Her-2/neu activation in response to P00533 tyrosine kinase inhibition in colon cancer cell lines . BACKGROUND : Tyrosine kinase receptors of the ErbB family have become promising targets for anti-neoplastic drugs , but mechanisms of resistance are incompletely understood . To investigate such pathways , we applied a small-molecule , selective P00533 inhibitor , DB00530 , to three well-characterized colon cancer cell lines and studied the alterations of expression and activation of receptors in the erbB family . METHODS : MTT assays were performed to determine the IC(50)s of GEO , FET , and HCT 116 human colorectal cancer cell lines treated with DB00530 . Plated cells were then exposed to either DB01093 control or 7 microm of DB00530 for treatment durations of 1 , 3 , 5 , 7 , 10 , 14 , 28 , and 56 days . Cell lysates were evaluated by Western blotting , evaluating both total and phosphorylated levels of P00533 , Her-2/neu , and erbB-3 . RESULTS : IC(50) values for GEO , FET , and HCT 116 cell lines exposed to DB00530 were 12.0 , 16.0 , and greater than 100 microm , respectively . In all treated cell lines , DB00530 diminished P00533 activation but did not affect total expression compared with controls . In contrast , Her-2/neu activation was increased in all cell lines . These changes in P00533 and Her-2/neu were identified within 24 h but peaked later in the treatment cycle . ErbB-3 expression and activation did not follow a consistent pattern between cell lines . CONCLUSIONS : Inhibition of P00533 led to increased activation of Her-2/neu . This result suggests a possible mechanism by which cells might escape the proapoptotic signals resulting from P00533 blockade . Our findings suggest concurrent inhibition of multiple members of the erbB family may yield stronger apoptotic responses than single receptor blockade alone . Clinical trials in thrombolytic therapy , Part 2 : The open-artery hypothesis and RAPID-1 and RAPID-2 . The open-artery hypothesis as supported by thrombolytic study results is discussed . The open-artery hypothesis states that survival after acute myocardial infarction ( AMI ) is maximized by achieving early and sustained patency of the infarct-related artery . However , two large multicenter trials did not detect any difference in mortality between patients given alteplase and patients given streptokinase , despite previous evidence that alteplase led to earlier recanalization of infarct-related arteries . The Global Utilization of DB00086 and Tissue P00747 Activator for Occluded Coronary Arteries ( GUSTO-1 ) trial suggested that early and complete patency is essential for short-term survival after AMI . Subsequent observations indicated that an open infarct-related artery at the time of hospital discharge is associated with improved long-term survival . In the DB00015 Angiographic Phase II International Dose-Finding ( RAPID-1 ) trial , complete patency was more frequent in patients who received a double-bolus regimen of reteplase than in patients who received standard-dose alteplase . Similar results were obtained in the DB00015 versus DB00009 Patency Investigation during Myocardial Infarction ( RAPID-2 ) trial , which compared the same double-bolus reteplase regimen with an accelerated regimen of alteplase . In both RAPID studies , mortality was lower and other outcomes were more favorable in reteplase recipients . DB00015 seems more likely to produce normal blood flow soon after AMI than either standard-dose or accelerated alteplase and may be associated with a lower mortality rate . This lends further support to the open-artery hypothesis . A curated database of miRNA mediated feed-forward loops involving MYC as master regulator . BACKGROUND : The MYC transcription factors are known to be involved in the biology of many human cancer types . But little is known about the Myc/microRNAs cooperation in the regulation of genes at the transcriptional and post-transcriptional level . METHODOLOGY/PRINCIPAL FINDINGS : Employing independent databases with experimentally validated data , we identified several mixed microRNA/Transcription Factor Feed-Forward Loops regulated by Myc and characterized completely by experimentally supported regulatory interactions , in human . We then studied the statistical and functional properties of these circuits and discussed in more detail a few interesting examples involving Q01094 , P60484 , P06400 and P15692 . CONCLUSIONS/SIGNIFICANCE : We have assembled and characterized a catalogue of human mixed Transcription Factor/microRNA Feed-Forward Loops , having Myc as master regulator and completely defined by experimentally verified regulatory interactions . iSubgraph : integrative genomics for subgroup discovery in hepatocellular carcinoma using graph mining and mixture models . The high tumor heterogeneity makes it very challenging to identify key tumorigenic pathways as therapeutic targets . The integration of multiple omics data is a promising approach to identify driving regulatory networks in patient subgroups . Here , we propose a novel conceptual framework to discover patterns of miRNA-gene networks , observed frequently up- or down-regulated in a group of patients and to use such networks for patient stratification in hepatocellular carcinoma ( HCC ) . We developed an integrative subgraph mining approach , called iSubgraph , and identified altered regulatory networks frequently observed in HCC patients . The miRNA and gene expression profiles were jointly analyzed in a graph structure . We defined a method to transform microarray data into graph representation that encodes miRNA and gene expression levels and the interactions between them as well . The iSubgraph algorithm was capable to detect cooperative regulation of miRNAs and genes even if it occurred only in some patients . Next , the miRNA-mRNA modules were used in an unsupervised class prediction model to discover HCC subgroups via patient clustering by mixture models . The robustness analysis of the mixture model showed that the class predictions are highly stable . Moreover , the Kaplan-Meier survival analysis revealed that the HCC subgroups identified by the algorithm have different survival characteristics . The pathway analyses of the miRNA-mRNA co-modules identified by the algorithm demonstrate key roles of Myc , Q01094 , let-7 , P01137 , P01375 and P00533 in HCC subgroups . Thus , our method can integrate various omics data derived from different platforms and with different dynamic scales to better define molecular tumor subtypes . iSubgraph is available as MATLAB code at http://www.cs.umd.edu/~ozdemir/isubgraph/ . Endothelin@25 - new agonists , antagonists , inhibitors and emerging research frontiers : IUPHAR Review 12 . Since the discovery of endothelin ( ET ) -1 in 1988 , the main components of the signalling pathway have become established , comprising three structurally similar endogenous 21-amino acid peptides , ET-1 , P20800 and P14138 , that activate two GPCRs , P25101 and ETB . Our aim in this review is to highlight the recent progress in ET research . The ET-like domain peptide , corresponding to prepro-ET-193-166 , has been proposed to be co-synthesized and released with ET-1 , to modulate the actions of the peptide . ET-1 remains the most potent vasoconstrictor in the human cardiovascular system with a particularly long-lasting action . To date , the major therapeutic strategy to block the unwanted actions of ET in disease , principally in pulmonary arterial hypertension , has been to use antagonists that are selective for the P25101 receptor ( ambrisentan ) or that block both receptor subtypes ( DB00559 ) . DB08932 represents the next generation of antagonists , being more potent than DB00559 , with longer receptor occupancy and it is converted to an active metabolite ; properties contributing to greater pharmacodynamic and pharmacokinetic efficacy . A second strategy is now being more widely tested in clinical trials and uses combined inhibitors of ET-converting enzyme and neutral endopeptidase such as SLV306 ( daglutril ) . A third strategy based on activating the ETB receptor , has led to the renaissance of the modified peptide agonist IRL1620 as a clinical candidate in delivering anti-tumour drugs and as a pharmacological tool to investigate experimental pathophysiological conditions . Finally , we discuss biased signalling , epigenetic regulation and targeting with monoclonal antibodies as prospective new areas for ET research . DB01197 inhibits neutrophil synthesis of leukotriene B4 in vitro and in vivo . The aim of this investigation was to determine the effects of metalloproteinase inhibitors on leukotriene ( LT ) A4 hydrolase in human neutrophil cytosol and to examine the effects of captopril on intact neutrophils in vitro and in vivo . Cytosolic fractions were assayed for P09960 hydrolase and P09917 activity in the presence or absence of inhibitors . Only bestatin , 1,10-O-phenanthroline , captopril and fosinoprilat demonstrated significant effects . The IC50 of captopril and fosinoprilat for P09960 hydrolase activity were 500 microM and 1 mM , respectively . No inhibition of P09917 activity in cytosolic fractions was detected . The effect of captopril was only minimally reversed by ZnSO4 . The IC50 of captopril for inhibition of LTB4 synthesis in intact neutrophils was 63 microM . Furthermore , 5-HETE production in intact cells was diminished 25.3 +/- 8.5 % in the presence of 1 mM captopril . Oral captopril inhibited stimulated LTB4 release by subsequently isolated neutrophils by 48.1 +/- 5.6 % and 5-HETE release by 43.2 +/- 5.5 % . Thus , captopril is an inhibitor of LTB4 synthesis in neutrophils in vitro and in vivo . However , there are differences between the potency of this drug as assessed in cytosol and intact cell studies . This study significantly extends previous reports in that it demonstrates that captopril is a more potent inhibitor of LTB4 synthesis in intact neutrophils than in cytosol and in that it demonstrates an inhibitory effect of captopril on synthesis of LTB4 by neutrophils exposed to captopril in vivo . Guanylate cyclase C-mediated antinociceptive effects of linaclotide in rodent models of visceral pain . BACKGROUND DB08890 is a novel , orally administered investigational drug currently in clinical development for the treatment of constipation-predominant irritable bowel syndrome ( IBS-C ) and chronic idiopathic constipation . Visceral hyperalgesia is a major pathophysiological mechanism in IBS-C . Therefore , we investigated the anti-nociceptive properties of linaclotide in rodent models of inflammatory and non-inflammatory visceral pain and determined whether these pharmacological effects are linked to the activation of guanylate cyclase C ( P25092 ) . METHODS Orally administered linaclotide was evaluated in non-inflammatory acute partial restraint stress ( PRS ) and acute water avoidance stress ( P42768 ) models in Wistar rats , and in a trinitrobenzene sulfonic acid ( TNBS ) -induced inflammatory model in Wistar rats and P25092 null mice . KEY RESULTS In TNBS-induced colonic allodynia , linaclotide significantly decreased the number of abdominal contractions in response to colorectal distension without affecting the colonic wall elasticity change in response to distending pressures after TNBS . However , linaclotide had no effect on visceral sensitivity under basal conditions . In addition , linaclotide significantly decreased colonic hypersensitivity in the PRS and P42768 models . In wild type ( wt ) and P25092 null mice , the instillation of TNBS induced similar hyperalgesia and allodynia . However , in post-inflammatory conditions linaclotide significantly reduced hypersensitivity only in wt mice , but not in P25092 null mice . CONCLUSIONS & INFERENCES These findings indicate that linaclotide has potent anti-nociceptive effects in several mechanistically different rodent models of visceral hypersensitivity and that these pharmacological properties of linaclotide are exerted through the activation of the P25092 receptor . Therefore , linaclotide may be capable of decreasing abdominal pain in patients suffering from IBS-C . Dexamethasone reverses adrenalectomy-induced neuronal de-differentiation in midbrain raphe-hippocampus axis . Differentiation leads to specific morphological and biochemical characteristics . We examined whether epigenetic factors ( e.g. , glucocorticoids ) are required to maintain neuronal differentiation in the adult brain . In the midbrain , adrenalectomy ( P10109 ) ( 1-2 wk ) reduced the size of tryptophan hydroxylase ( WH ) -immunoreactive ( IR ) neurons . P10109 rats exposed to short-term ( 24-72-h ) dexamethasone ( ST-DEX ) in the drinking saline ( 10 mg/l ) showed an increase in WH protein , somal area and dendritic size of WH-IR neurons . In the hippocampus , P10109 for 2-3 mo ( long-term ; LT ) reduced Nissl staining , calbindin ( DB09061 ) -IR and P08908 receptor mRNA in the granular cell layer , and the size of the molecular layer and its DB09061 -IR dendrites . Small vimentin ( Vim ) -IR glial cells emerged in the granular layer . ST-DEX after LT- P10109 rapidly induced a recovery of P08908 mRNA , Nissl labeling and DB09061 -IR in the granule cell layer . In the molecular layer , there was an increase in the area and in the number of DB09061 -IR dendrites . Furthermore , the Vim-IR glial cells were enlarged in size and branching . The rate of cell proliferation was studied in these animals . Immunostaining with antibodies against proliferating cell nuclear antigen ( P12004 ) and use of bromouridine argue against enhanced neurogenesis after ST-DEX in LT- P10109 . We propose that glucocorticoids induce and maintain differentiation of serotonergic and DB09061 -IR neurons in the midbrain-hippocampal axis . A neuronotrophic role for the glial P08908 receptor is suggested . Clinical utility of vilazodone for the treatment of adults with major depressive disorder and theoretical implications for future clinical use . BACKGROUND : DB06684 is the latest approved antidepressant available in the United States . Its dual mechanism of action combines the inhibition of serotonin transporters while simultaneously partially agonizing serotonin-1a ( P08908 ) receptors . This combined activity results in serotonin facilitation across the brain 's serotonergic pathways , which has been termed by the authors as that of a serotonin partial agonist and reuptake inhibitor , or SPARI . OBJECTIVE : The authors to review laboratory , animal model data , and human trial data to synthesize a working theory regarding the mechanism of antidepressant action of this agent and regarding its potential for additional indications . METHODS : A MEDLINE and Internet search was conducted and the resultant evidence reviewed . RESULTS : DB06684 has randomized , controlled empirical data which has garnered it an approval for treating major depressive disorder . It combines two well-known pharmacodynamic mechanisms of serotonergic action into a novel agent . Although no head-to-head studies against other antidepressants are published , the efficacy data for vilazodone appears comparable to other known antidepressants , with associated gastrointestinal side effects similar to serotonin selective reuptake inhibitor and serotonin norepinephrine reuptake inhibitor antidepressants , but potentially with a lower incidence of sexual side effects and weight gain . DISCUSSION : As a new option for the treatment of major depressive disorder , vilazodone , due to its unique SPARI mechanism of action , may hold promise for patients who can not tolerate or have not responded to previous antidepressant monotherapies . Additionally , its use may extend to the treatment of other mental health conditions similar to those treated by serotonin selective reuptake inhibitors . Thirteen type I loci from HSA4q , HSA6p , HSA7q and HSA12q were comparatively Q5TCZ1 -mapped in four river buffalo and sheep chromosomes . Thirteen goat BAC clones containing coding sequences from HSA7 , HSA12q , HSA4 and HSA6p were fluorescence in situ mapped to river buffalo ( Bubalus bubalis , BBU ) and sheep ( Ovis aries , OAR ) R-banded chromosomes . The following type I loci were mapped : P03999 to BBU8q32 and OAR4q32 , P35523 to BBU8q34 and OAR4q34 , P17936 to BBU8q24 and OAR4q27 , KRT to BBU4q21 and OAR 3q21 , P01579 to BBU4q23 and OAR3q23 , IGF1 to BBU4q31 and OAR3q31 , P30968 to BBU7q32 and OAR6q32 , P55157 to BBU7q21 and OAR6q15 , P35913 to BBU7q36 and OAR6q36 , BF to BBU2p22 and OAR20q22 , P05305 to BBU2p24 and OAR20q24 , P08263 to BBU2p22 and OAR20q22 , OLADRB ( MHC ) to BBU2p22 and OAR20q22 . All mapped loci appeared to be located on homologous chromosomes and chromosome bands in both bovids . Comparison between gene orders in bovid ( BBU and OAR ) and human ( HSA ) chromosomes revealed complex rearrangements , especially between BBU7/OAR6 and HSA4 , as well as between BBU2p/OAR20 and HSA6p . DB08932 : first global approval . DB08932 ( Opsumit® ) is a novel dual endothelin receptor antagonist ( ERA ) with sustained receptor binding properties developed by Actelion Pharmaceuticals Ltd . In October 2013 , oral macitentan 10 mg once daily received its first global approval in the US , followed closely by Canada , for the treatment of pulmonary arterial hypertension ( PAH ) . The drug has also received a positive opinion in the EU from the Committee for Medicinal Products for Human Use for the treatment of PAH , and is under regulatory review in several other countries for the same indication . Endothelin ( ET ) -1 influences pathological changes via two ET receptor subtypes ( P25101 and ETB ) , to which it binds with high affinity . ET-1 is implicated in several forms of vascular disease making it a valid target for the treatment of pulmonary vascular diseases such as PAH . Clinical development is underway for other indications , including Eisenmenger syndrome , ischaemic digital ulcers secondary to systemic sclerosis , and glioblastoma . DB08932 was also evaluated in idiopathic pulmonary fibrosis ; however , a phase 2 trial did not meet its primary endpoint and further investigation in this indication was discontinued . DB08932 was developed by modifying the structure of DB00559 in the search for an optimal dual ERA with improved efficacy and tolerability compared with other ERAs . This article summarizes the milestones in the development of macitentan leading to this first approval for PAH . 17 DB00783 -mediated growth inhibition of MDA-MB-468 cells stably transfected with the estrogen receptor : cell cycle effects . P03372 ( ER ) -negative MDA-MB-468 human breast cancer cells were stably transfected with wild-type human ER and utilized as a model for investigating estrogen- and aryl hydrocarbon ( Ah ) -responsiveness . Treatment of the stably transfected cells with 10 nM 17 beta-estradiol ( E2 ) resulted in a significant inhibition ( > 60 % ) of cell proliferation and DNA synthesis , which was blocked by 10(-7) M ICI 182 780 . Analysis by flow cytometry indicated that treatment with E2 increased the percentage of cells in G0/ P55008 ( from 68.8 to 89.4 ) and decreased cells in S ( from 18.4 to 3.4 ) and G2/M ( from 12.8 to 7.2 ) phases of the cell cycle . The effects of E2 on the major cyclins , cyclin-dependent kinases and cyclin-dependent kinase inhibitors , retinoblastoma protein ( RB ) , Q01094 , and cyclin-dependent kinase activities were also investigated in the stably transfected MDA-MB-468 cells . The results demonstrated that the growth inhibitory effects of 10(-8) M E2 in ER stably transfected MDA-MB-468 cells were associated with modulation of several factors required for cell cycle progression and DNA synthesis , including significant induction of the cyclin-dependent kinase inhibitor p21cip-1 ( > 4-fold increase after 12 h ) and decreased Q01094 and P12004 protein levels . These results show that the growth-inhibitory effects of E2 in the stably transfected cells were due to multiple factors which result in growth arrest in G0/ P55008 and inhibition of DNA synthesis . A mechanism-based model for the population pharmacokinetics of free and bound aflibercept in healthy subjects . AIM : DB08885 ( P15692 - Q8NHU6 ) , a novel anti-angiogenic agent that binds to P15692 , has been investigated for the treatment of cancer . The aim of this study was to develop a mechanism-based pharmacokinetic ( PK ) model for aflibercept to characterize its binding to P15692 and its PK properties in healthy subjects . METHODS : Data from two phase I clinical studies with aflibercept administered as a single intravenous infusion were included in the analysis . Free and bound aflibercept concentration-time data were analysed using a nonlinear mixed-effects modelling approach with MONOLIX 3.1 . RESULTS : The best structural model involved two compartments for free aflibercept and one for bound aflibercept , with a Michaelis-Menten type binding of free aflibercept to P15692 from the peripheral compartment . The typical estimated clearances for free and bound aflibercept were 0.88 l day(-1) and 0.14 l day(-1) , respectively . The central volume of distribution of free aflibercept was 4.94 l . The maximum binding capacity was 0.99 mg day(-1) and the concentration of aflibercept corresponding to half of maximum binding capacity was 2.91 µg ml(-1) . Interindividual variability of model parameters was moderate , ranging from 13.6 % ( V(max) ) to 49.8 % ( Q ) . CONCLUSION : The present PK model for aflibercept adequately characterizes the underlying mechanism of disposition of aflibercept and its nonlinear binding to P15692 . DB01197 inhibits the fluorescence development associated with glycation of proteins . Albumin and immunoglobulin G ( IgG ) show increased visible fluorescence in diabetic patients , IgG fluorescence being correlated with the presence of diabetic retinopathy . DB01197 , an angiotensin converting enzyme ( P12821 ) inhibitor , has free radical scavenging ability , attributable to its thiol group . We compared the scavenging effect of captopril ( at doses between 0.5 and 100 microM ) with perindoprilat , enalapril and enalaprilat ( P12821 inhibitors without scavenging ability ) and two thiol-containing compounds , mercaptopropionylglycine ( P29372 ) and DB06151 ( Q9C000 ) ( scavengers with no effect on P12821 ) . Three systems were used to generate visible fluorescence in albumin and IgG ; glycation , exposure to copper/hydrogen peroxide and gamma radiation . All three thiol-containing compounds inhibited fluorescence development in IgG and albumin , when fluorescence was generated by glycation or gamma radiation . Other P12821 inhibitors had no effect with IgG . Enalapril and perindoprilat showed less effect than captopril with albumin ; enalaprilat had no effect . No compound had any effect on fluorescence generation by copper/hydrogen peroxide . DB01197 may have an additional antioxidant effect compared to other P12821 inhibitors . Estrogen upregulates endothelial nitric oxide synthase gene expression in fetal pulmonary artery endothelium . NO , produced by endothelial NO synthase ( P29474 ) , is a key mediator of pulmonary vasodilation during cardiopulmonary transition at birth . The capacity for NO production is maximal at term because pulmonary P29474 expression increases during late gestation . Since fetal estrogen levels rise markedly during late gestation and there is indirect evidence that the hormone enhances nonpulmonary NO production in adults , estrogen may upregulate P29474 in fetal pulmonary artery endothelium . Therefore , we studied the direct effects of estrogen on P29474 expression in ovine fetal pulmonary artery endothelial cells ( PAECs ) . DB00783 caused a 2.5-fold increase in NOS enzymatic activity in PAEC lysates . This effect was evident after 48 hours , and it occurred in response to physiological concentrations of the hormone ( 10(-10) to 10(-6) mol/L ) . The increase in NOS activity was related to an upregulation in P29474 protein expression , and P29474 mRNA abundance was also enhanced . P03372 antagonism with DB00947 completely inhibited estrogen-mediated P29474 upregulation , indicating that estrogen receptor activation is necessary for this response . In addition , immunocytochemistry revealed that fetal PAECs express estrogen receptor protein . Furthermore , transient transfection assays with a specific estrogen-responsive reporter system have demonstrated that the endothelial estrogen receptor is capable of estrogen-induced transcriptional transactivation . Thus , estrogen upregulates P29474 gene expression in fetal PAECs through the activation of PAEC estrogen receptors . This mechanism may be responsible for pulmonary P29474 upregulation during late gestation , thereby optimizing the capacity for NO-mediated pulmonary vasodilation at birth . Growth factors and their receptors : new targets for prostate cancer therapy . Stimulation of the signal transduction pathway of the epidermal growth-factor receptor ( P00533 ) tyrosine kinase family of receptors in tumor cells enhances cellular proliferation , prevents apoptosis , and promotes tumor-cell mobility , adhesion , and invasion . Therapeutic approaches used to target the P00533 and its signal transduction cascade include ( 1 ) monoclonal antibodies ( eg , cetuximab [ IMC-C225 ] ) directed against the extracellular binding domain of the receptor ; and ( 2 ) trastuzumab , a monoclonal antibody binding to the P04626 receptor ; immunotoxin conjugates use an antibody directed against P00533 joined to a cell toxin . All are in clinical trials for a number of cancers , including prostate cancer . Antisense strategies are in preclinical development . Low-molecular-weight inhibitors of the P00533 tyrosine kinase also in clinical development include DB00530 , PD182905 , PKI-166 , DB05424 , and ZD1839 . ZD1839 has shown encouraging results in patients with prostate cancer in phase 1 trials. mn Target-based agents against ErbB receptors and their ligands : a novel approach to cancer treatment . The ErbB receptors and their cognate ligands that belong to the epidermal growth factor ( P01133 ) family of peptides are involved in the pathogenesis of different types of carcinomas . In fact , the ErbB receptors and the P01133 -like growth factors are frequently expressed in human tumors . These proteins form a complex system that regulates the proliferation and the survival of cancer cells . Therefore , ErbB receptors and their ligands might represent suitable targets for novel therapeutic approaches in human carcinomas . In this regard , different target-based agents that are directed against the ErbB receptors have been developed in the past two decades . One of these compounds , the humanized anti-ErbB-2 monoclonal antibody trastuzumab has been approved for the treatment of patients with metastatic breast cancer . The anti- P01133 receptor ( P00533 ) antibody C225 , as well as P00533 tyrosine kinase inhibitors ZD1839 and DB00530 are currently in phase III clinical development . Several other ErbB tyrosine kinase inhibitors are in phase I/II studies . These compounds have generally been shown to have an acceptable toxicity profile and promising anti-tumor activity in heavily pretreated patients . The mechanisms of action of these compounds , as well as the potential therapeutic strategies to improve their efficacy are discussed in this review with particular regard to the combinations of anti-ErbB agents with cytotoxic drugs , or combinations of different ErbB-targeting agents . DB00559 , an endothelin receptor antagonist , ameliorates collagen-induced arthritis : the role of P01375 -α in the induction of endothelin system genes . OBJECTIVE : Endothelins ( ETs ) are involved in several inflammatory events . The present study investigated the efficacy of DB00559 , a dual P25101 /ETB receptor antagonist , in collagen-induced arthritis ( CIA ) in mice . TREATMENT : CIA was induced in DBA/1J mice . Arthritic mice were treated with DB00559 ( 100 mg/kg ) once a day , starting from the day when arthritis was clinically detectable . METHODS : CIA progression was assessed by measurements of visual clinical score , paw swelling and hypernociception . Histological changes , neutrophil infiltration and pro-inflammatory cytokines were evaluated in the joints . Gene expression in the lymph nodes of arthritic mice was evaluated by microarray technology . PreproET-1 mRNA expression in the lymph nodes of mice and in peripheral blood mononuclear cells ( PBMCs ) was evaluated by real-time PCR . The differences were evaluated by one-way Q9UNW9 or Student 's t test . RESULTS : Oral treatment with DB00559 markedly ameliorated the clinical aspects of CIA ( visual clinical score , paw swelling and hyperalgesia ) . DB00559 treatment also reduced joint damage , leukocyte infiltration and pro-inflammatory cytokine levels ( IL-1β , TNFα and Q16552 ) in the joint tissues . Changes in gene expression in the lymph nodes of arthritic mice returned to the levels of the control mice after DB00559 treatment . PreproET mRNA expression increased in PBMCs from rheumatoid arthritis ( RA ) patients but returned to basal level in PBMCs from patients under anti- P01375 therapy . In-vitro treatment of PBMCs with TNFα upregulated ET system genes . CONCLUSION : These findings indicate that ET receptor antagonists , such as DB00559 , might be useful in controlling RA . Moreover , it seems that ET mediation of arthritis is triggered by TNFα . Drug evaluation : DB06684 -- a combined SSRI and P08908 partial agonist for the treatment of depression . DB06684 is a combined selective serotonin reuptake inhibitor ( SSRI ) and a 5-HT(1A) receptor partial agonist that is being developed by Clinical Data Inc for the treatment of depression . In preclinical studies , vilazodone compared favorably to other antidepressants such as paroxetine and fluoxetine . Orally administered vilazodone inhibited ultrasonic vocalization in the rat after electrical foot shock ( a model of anxiolytic activity ) . Yet , in the forced swimming test model of depression in rats , vilazodone administered intraperitoneally was active at 1 mg/kg but not at 3 or 10 mg/kg . During clinical trials , vilazodone completely abolished O75628 sleep for 8 h and demonstrated antidepressant efficacy that was equal to that of current antidepressant therapeutics . The author concludes that the success of vilazodone as an effective antidepressant agent will depend on whether the drug can produce a more rapid antidepressant effect than other SSRI agents , or if specific genetic markers of patients can be associated with clinical efficacy . Negative pressure accelerated monolayer keratinocyte healing involves Cdc42 mediated cell podia formation . BACKGROUND : Negative-pressure wound therapy ( NPWT ) is developed to facilitate wound healing at controlled subatmospheric pressures in modern medicine . Molecular mechanism for this therapy is still undefined . OBJECTIVE : This study highlights the localization and time-course of the cell division control protein 42 ( Cdc42 ) in the cell membrane at ambient pressure ( AP ) and negative pressures of 75mmHg ( NP75 ) , 125mmHg ( NP125 ) and 175mmHg ( NP175 ) . METHODS : The prepared cells were cultured in a negative pressure incubator with the same O2 and CO2 tensions at the four different pressures . The effective time , complete wound closure time , cell volume , cell viability , and the fluorescence of proliferating cell nuclear antigens ( P12004 ) and actins were evaluated in cells at different pressures . Wound-healing process and Cdc42 fluorescence were examined in cells with the knockdown of Cdc42 . Cdc42 pathway proteins in cell membranes were analyzed after incubation at different pressures for 6 and 12h . RESULTS : The cells at NP125 had less wound closure time and obvious cell podia . Similar P12004 fluorescent intensity was observed in cells at different pressures . The Cdc42 , neural P42768 , and actin expression increased significantly ( p < 0.05 ) in plasma membranes of cells at NP125 for 12h . The knockdown of active Cdc42 resulted in the absence of Cdc42 expression at the cell leading edge . CONCLUSIONS : The activation and localization of Cdc42 pathway proteins in the cell membrane are involved in the cell podia formation in keratinocytes at NP125 . NPWT may facilitate cell migration to accelerate wound healing . Role of endothelin in the maintenance of blood pressure in conscious rats with chronic heart failure . Acute effects of the endothelin receptor antagonist Ro 47-0203 ( DB00559 ) . BACKGROUND : Endothelin ( ET ) is a potent vasoconstrictor , and its concentration is increased in patients with heart failure . The purpose of this study was to investigate the role of endothelin in heart failure by use of a rat model . METHODS AND RESULTS : Experiments were performed on rats at 1 through 16 weeks after sham operation or coronary artery ligation . Rats with left ventricular end-diastolic pressures > 15 mm Hg were considered to have chronic heart failure ( CHF ) , while the others were considered to have uncomplicated myocardial infarction ( MI ) . There were increased ET-1 concentrations in CHF rats at weeks 1 to 16 ( Sham , 20 +/- 0.5 pg/mL , n = 45 ; CHF , 31 +/- 2 pg/mL , n = 50 ; P < .001 ) and transient increases in P14138 concentrations at week 1 in both the MI and CHF groups . There were no significant increases in big ET-1 concentrations , suggesting an increased conversion of ET-1 from big ET-1 in the CHF group . At weeks 2 through 8 , oral administration of the mixed ( P25101 and ETB ) endothelin receptor antagonist DB00559 significantly decreased mean arterial pressure in conscious CHF rats , an effect that increased over time . Furthermore , DB00559 had an additive effect to the angiotensin-converting enzyme inhibitor cilazapril . CONCLUSIONS : Endothelin plays a role in the maintenance of blood pressure in CHF rats , as evidenced by the significant reduction in mean arterial pressure after oral administration of DB00559 . Therefore , endothelin antagonists may be useful therapeutic agents in the treatment of CHF . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . DB08932 slows down the dermal fibrotic process in systemic sclerosis : in vitro findings . Systemic sclerosis ( or scleroderma ) is an autoimmune disease characterized by skin and internal organ fibrosis , caused by microvascular dysfunction . The microvascular damage seems to be a consequence of an endothelial autoimmune response , followed by activation of the inflammatory cascade and massive deposition of collagen . P05305 ( ET-1 ) contributes to the inflammatory and fibrotic processes by increasing the concentration of pro-inflammatory and pro-fibrotic cytokines , and it is considered one of the most relevant mediators of vascular damage in scleroderma . It is indeed found in very high concentration in serum of sclerodermic patients . Moreover , in these pathological conditions there is an increased expression of ET-1 receptors ( P25101 and ETB ) , which mediate the detrimental action of ET-1 , and often a change of P25101 /ETB ratio . The aim of the present study is to evaluate the in vitro effect of macitentan , an orally active tissue-targeting dual endothelin receptor antagonist , and its major metabolite ( ACT-132577 ) on alpha smooth muscle actin ( alphaSMA ) expression , evaluated on dermal fibroblasts from healthy subjects and on dermal fibroblasts from lesional and non-lesional skin from sclerodermic patients . The combination of macitentan and its major metabolite reduced the levels of & #945;SMA after 48 h in sclerodermic fibroblasts from lesional skin . No relevant changes in & #945;SMA levels were found in fibroblasts from non-lesional skin , whose behavior is similar to that of dermal fibroblasts from healthy patients . Regulation and therapeutic targeting of peptide-activated receptor guanylyl cyclases . Cyclic GMP is a ubiquitous second messenger that regulates a wide array of physiologic processes such as blood pressure , long bone growth , intestinal fluid secretion , phototransduction and lipolysis . Soluble and single-membrane-spanning enzymes called guanylyl cyclases ( GC ) synthesize cGMP . In humans , the latter group consists of P16066 , P20594 , P25092 , GC-E and P51841 , which are also known as P16066 , P20594 , StaR , Ret1-GC and Ret2-GC , respectively . Membrane GCs are activated by peptide ligands such as atrial natriuretic peptide ( P01160 ) , B-type natriuretic peptide ( DB04899 ) , P23582 ( P09543 ) , guanylin , uroguanylin , heat stable enterotoxin and GC-activating proteins . DB04899 and carperitide are clinically approved peptide-based drugs that activate P16066 . CD-NP is an experimental heart failure drug that primarily activates P20594 but also activates P16066 at high concentrations and is resistant to degradation . Inactivating mutations in P20594 cause acromesomelic dysplasia type Maroteaux dwarfism and chromosomal mutations that increase P09543 concentrations are associated with Marfanoid-like skeletal overgrowth . Pump-based P09543 infusions increase skeletal growth in a mouse model of the most common type of human dwarfism , which supports P09543 / P20594 -based therapies for short stature diseases . DB08890 is a peptide activator of P25092 that stimulates intestinal motility and is in late-stage clinical trials for the treatment of chronic constipation . This review discusses the discovery of cGMP , guanylyl cyclases , the general characteristics and therapeutic applications of P16066 , P20594 and P25092 , and emphasizes the regulation of transmembrane guanylyl cyclases by phosphorylation and DB00171 . P05121 plays a protective role in CCl4-induced hepatic fibrosis in mice : role of hepatocyte division . P00747 activator inhibitor-1 ( P05121 ) is an acute phase protein that has been shown to play a role in experimental fibrosis caused by bile duct ligation ( BDL ) in mice . However , its role in more severe models of hepatic fibrosis ( e.g. , carbon tetrachloride ; CCl(4) ) has not been determined and is important for extrapolation to human disease . Wild-type or P05121 knockout mice were administered CCl(4) ( 1 ml/kg body wt ip ) 2x/wk for 4 wk . Plasma ( e.g. , transaminase activity ) and histological ( e.g. , Sirius red staining ) indexes of liver damage and fibrosis were evaluated . Proliferation and apoptosis were assessed by P12004 and TdT-mediated dUTP nick-end labeling ( TUNEL ) staining , respectively , as well as by indexes of cell cycle ( e.g. , p53 , cyclin D1 ) . In contrast to previous studies with BDL , hepatic fibrosis was enhanced in P05121 (-/-) mice after chronic CCl(4) administration . Indeed , all indexes of liver damage were elevated in P05121 (-/-) mice compared with wild-type mice . This enhanced liver damage correlated with impaired hepatocyte proliferation . A similar effect on proliferation was observed after one bolus dose of CCl(4) , without concomitant increases in liver damage . Under these conditions , a decrease in phospho-p38 , coupled with elevated p53 protein , was observed ; these results suggest impaired proliferation and a potential G(1)/S cell cycle arrest in P05121 (-/-) mice . These data suggest that P05121 may play multiple roles in chronic liver diseases , both protective and damaging , the latter mediated by its influence on inflammation and fibrosis and the former via helping maintain hepatocyte division after an injury . Successful thrombolysis of a stroke with a pulmonary embolism in a young woman . BACKGROUND : Paradoxical embolism is a rare event , accounting for < 2 % of all arterial emboli . The diagnosis is often difficult , and consequences for the patient can be severe . CASE REPORT : We describe the case of a 35-year-old female physician who presented to our Emergency Department ( ED ) in severe hemodynamic compromise , with an altered level of consciousness and major expressive aphasia 1 day after undergoing a leg varicosal stripping procedure under regional anesthesia . She was successfully thrombolyzed with 0.9 mg/kg of Recombinant Tissue P00747 Activator ( rtPA , DB00009 ) and had a full recovery . CONCLUSION : To our knowledge , this is the first description of a case of massive pulmonary embolism associated with a paradoxical stroke related to patent foramen ovale that was thrombolyzed for both conditions with a " neurological dose " of rtPA . Although thrombolysis was completely successful in this case , indications and contraindications should be thoroughly respected . A more conservative approach with anticoagulation , or a more aggressive approach with surgical thrombectomy , can each potentially have a place in particular cases . Intra-arterial catheter-directed thrombolysis and percutaneous embolectomy are additional options to be considered when available , especially if there are contraindications for systemic thrombolysis . Pharmacologic characterization of endothelin receptor responses in the isolated perfused rat lung . Endothelin receptor subtypes were characterized in isolated perfused rat lungs using the peptide P25101 -receptor antagonists BQ 610 and BQ 123 , the nonpeptide mixed P25101 -/ETB-receptor antagonist DB00559 , and the ETB-receptor agonist IRL 1620 . Intra-arterial injection of 1 nmol IRL 1620 caused an enhanced reduction in pulmonary conductance compared with 1 nmol endothelin ( ET-1 ) or 0.33 nmol IRL 1620 . Pretreatment of lungs with BQ 610 , BQ 123 , or DB00559 aggravated the bronchoconstriction induced by 1 nmol ET-1 so that it was comparable to that induced by 1 nmol IRL 1620 . Although perfusion with 1 nmol IRL 1620 had only minor effects on vascular conductance , 1 nmol ET-1 caused a marked decrease in this parameter . This vasonconstriction was prevented by BQ 610 , BQ 123 , or DB00559 . High concentrations of the stable prostacyclin metabolite , 6-keto-PGF1 alpha , were found in the perfusate of lungs treated with 1 nmol IRL 1620 or 1 nmol ET-1 . The ET-1-induced release of 6-keto-PGF1 alpha was blocked by DB00559 , but not by BQ 610 . ET-1 , but not IRL 1620 , provoked the release of thromboxane B2 . The main effect of P25101 -receptor stimulation is vasoconstriction , whereas ETB-receptor stimulation causes bronchoconstriction . Both actions , however , are attenuated by the other receptor , i.e. , the P25101 -induced vasoconstriction is attenuated by ETB-receptor-induced release of vasodilators such as prostacyclin , whereas the ETB-receptor-induced bronchoconstriction is attenuated by an unknown P25101 -receptor-dependent bronchodilatory mechanism . Angiotensin converting enzyme inhibition in the postnatal rat results in decreased cell proliferation in the renal outer medulla . 1. Chronic angiotensin converting enzyme ( P12821 ) inhibition or AT1 antagonism during postnatal development in the rat has been shown to cause renal tubular and vascular damage , particularly in the outer medulla . 2 . The effects of P12821 inhibition were investigated at a stage of development before the renal outer medulla is fully established . 3 . Sprague-Dawley rat pups were given daily i.p. injections of either enalapril or saline from days 3-10 . At day 11 , kidneys were perfusion-fixed for either electron microscopy or immunocytochemistry . Sections were incubated in proliferating cell nuclear antigen ( P12004 ) antisera and the avidin-biotin immunoperoxidase method was used to detect an immunoreactive product , indicative of proliferating cells . 4 . Following enalapril treatment , the normal structural arrangement of the outer medulla was disrupted compared with controls . Cell proliferation ( P12004 -positive cells ) in the medullary rays was reduced in enalapril-treated kidneys compared with control kidneys . 5 . Thus , angiotensin II appears to be essential for normal tubular and vascular growth in postnatal renal development in the rat . P00533 tyrosine kinase inhibitors : application in non-small cell lung cancer . Despite treatment advances over the past decade , long-term survival for patients with non-small cell lung cancer ( NSCLC ) remains poor , and treatment options available after second-line therapy are limited . Increased understanding of cancer biology has led to the identification of several potential targets for treatment . The epidermal growth factor receptor ( P00533 ) belongs to a family of plasma membrane receptor tyrosine kinases that controls many important cellular functions , from growth and proliferation to cell death . This receptor is a particularly promising therapeutic target because it often is overexpressed in patients with NSCLC and has been implicated in the pathogenesis as well as the proliferation , invasion , and metastasis of lung cancer and other malignancies . New agents developed to inhibit P00533 function include small-molecule tyrosine kinase inhibitors , monoclonal antibodies to P00533 , and pan- P00533 inhibitors . Completed and ongoing clinical trials have shown that P00533 inhibitors have remarkable efficacy for patients with relapsed NSCLC . Among these , two phase 2 trials have shown that ZD1839 is effective when used as monotherapy . The response rates are comparable with those for docetaxel given in the second-line setting . Another phase 2 trial has shown that DB00530 is effective in the same setting . Data from phase 3 trials indicate that adding an P00533 tyrosine kinase inhibitor to chemotherapy does not provide an additional survival benefit , as compared with standard chemotherapy alone for first-line treatment of NSCLC . It appears that P00533 tyrosine kinase inhibitors are safe and well tolerated by patients with cancer . Further studies will elucidate how these new agents can best be used for NSCLC and other tumor types . Valproic acid suppresses cervical cancer tumor progression possibly via activating Notch1 signaling and enhances receptor-targeted cancer chemotherapeutic via activating somatostatin receptor type II . PURPOSE : We investigated the effects of the anti-epilepsy drug valproic acid ( DB00313 ) alone and in combination in treating cervical cancer . METHODS : DB00313 was investigated for its effects on cervical cancer Hela cell proliferation and tumor growth via in vitro and in vivo assays . RESULTS : DB00313 induce cell growth suppression and cell cycle arrest , with an increase of Notch1 that acts as a tumor suppressor and the change of other tumor-associated genes such as P38936 , p63 and P12004 . DB00313 was also found to induce cell morphological change , with an increase of certain cell transformation markers such as snail1 , snail2 and P19022 . Moreover , DB00313 could significantly up-regulate somatostatin receptor type II ( P30874 ) . Our in vivo study further demonstrated that DB00313 via inducing P30874 up-regulation extremely enhanced the anti-tumor ability of the P30874 -preferential cytotoxic COL- P61278 conjugate in xenografts . CONCLUSIONS : DB00313 could not only suppress tumor progression but also provide a novel promising therapeutic choice in combination with a receptor-targeted cytotoxic conjugate via activating the specific receptor . Somatostatin analogues inhibit cancer cell proliferation in an P30874 -dependent manner via both cytostatic and cytotoxic pathways . Somatostatin receptors ( SSTRs ) are inhibitory G-protein coupled receptors that are ubiquitously expressed in normal and cancer cells . Somatostatins ( P61278 ) are the natural ligands for SSTRs and act as inhibitory regulators of hormone secretion and proliferation . DB00104 and RC-160 ( vapreotide ) are two well tolerated P30874 / P35346 selective somatostatin analogues ( SSA ) that have been used in the treatment of cancers with mixed outcomes . Loss-of-expression of P30874 in tumor tissues has been suggested to correlate to tumor progressions and to the poor outcomes of somatostatin analogue treatment in certain clinical trials . In this study , exogenous human P30874 was overexpressed in two cancer cell lines , capan-2 cells and A549 cells , which had different profiles of endogenous SSTR expression . It was shown that overexpression of P30874 dramatically inhibited the proliferation of P30874 -positive and P30874 -negative cancer cells . Further growth inhibition of these cancer cells overexpressing P30874 was observed by application of octreotide/RC-160 in a dose-dependent fashion . In addition , immunoassay demonstrated that SSA/ P30874 inhibited proliferation via both cell cycle arresting and promoting apoptosis . The results suggested that P30874 could be a promising candidate for gene therapy for P30874 -positive and P30874 -negative tumors . The cellular level of P30874 might be a critical factor that could affect both tumor progression and the outcomes of somatostatin analogue treatment . Down-regulated P13569 During Aging Contributes to Benign Prostatic Hyperplasia . Benign prostatic hyperplasia ( BPH ) is a hyper-proliferative disease of the aging prostate ; however , the exact mechanism underlying the development of BPH remains incompletely understood . The present study investigated the possible involvement of the cystic fibrosis transmembrane conductance regulator ( P13569 ) , which has been previously shown to negatively regulate nuclear factor-κB ( NF-κB ) /cyclooxygenase 2 ( P35354 ) /prostaglandin E2 ( DB00917 ) pathway , in the pathogenesis of BPH . Our results showed decreasing P13569 and increasing P35354 expression in rat prostate tissues with aging . Furthermore , suppression of P13569 led to increased expression of P35354 and over-production of DB00917 in a normal human prostate epithelial cell line ( PNT1A ) with elevated NF-κB activity . DB00917 stimulated the proliferation of primary rat prostate stromal cells but not epithelial cells , with increased P12004 expression . In addition , the condition medium from PNT1A cells after inhibition or knockdown of P13569 promoted cell proliferation of prostate stromal cells which could be reversed by P35354 or NF-κB inhibitor . More importantly , the involvement of P13569 in BPH was further demonstrated by the down-regulation of P13569 and up-regulation of P35354 /NF-κB in human BPH samples . The present results suggest that P13569 may be involved in regulating DB00917 production through its negative regulation on NF-κB/ P35354 pathway in prostate epithelial cells , which consequently stimulates cell growth of prostate stromal cells . The overstimulation of prostate stromal cell proliferation by down-regulation of P13569 -enhanced DB00917 production and release during aging may contribute to the development of BPH . A multi-layer inference approach to reconstruct condition-specific genes and their regulation . An important topic in systems biology is the reverse engineering of regulatory mechanisms through reconstruction of context-dependent gene networks . A major challenge is to identify the genes and the regulations specific to a condition or phenotype , given that regulatory processes are highly connected such that a specific response is typically accompanied by numerous collateral effects . In this study , we design a multi-layer approach that is able to reconstruct condition-specific genes and their regulation through an integrative analysis of large-scale information of gene expression , protein interaction and transcriptional regulation ( transcription factor-target gene relationships ) . We establish the accuracy of our methodology against synthetic datasets , as well as a yeast dataset . We then extend the framework to the application of higher eukaryotic systems , including human breast cancer and Arabidopsis thaliana cold acclimation . Our study identified P09758 ( P09758 ) as a target gene for human breast cancer and discovered its regulation by transcription factors CREB , as well as NFkB . We also predict Q99661 is a target gene for ER-/ P04626 - breast cancer and is positively regulated by Q01094 . The predictions were further confirmed through experimental studies . AVAILABILITY : The implementation and detailed protocol of the layer approach is available at http://www.egr.msu.edu/changroup/Protocols/Three-layer % 20approach % 20 to % 20reconstruct % 20condition.html . Does a nonclassical signaling mechanism underlie an increase of estradiol-mediated gonadotropin-releasing hormone receptor binding in ovine pituitary cells ? DB00783 ( E2 ) is the major regulator of P30968 ( GnRHR ) gene expression and number during the periovulatory period ; however , the mechanisms underlying E2 regulation of the P30968 gene remain undefined . Herein , we find that E2 conjugated to BSA ( E2-BSA ) mimics the stimulatory effect of E2 on DB00644 binding in primary cultures of ovine pituitary cells . The time course for maximal DB00644 analog binding was similar for both E2 and E2-BSA . The ability of E2 and E2-BSA to increase DB00644 analog binding was blocked by the estrogen receptor ( ER ) antagonist DB00947 . Also , increased DB00644 analog binding in response to E2 and the selective P03372 agonist propylpyrazole triol was blocked by expression of a dominant-negative form of P03372 ( L540Q ) . Thus , membrane-associated P03372 is the likely candidate for mediating E2 activation of the P30968 gene . As DB02527 response element binding protein ( CREB ) is an established target for E2 activation in gonadotrophs , we next explored a potential role for this protein as an intracellular mediator of the E2 signal . Consistent with this possibility , adenoviral-mediated expression of a dominant-negative form of CREB ( A-CREB ) completely abolished the ability of E2 to increase DB00644 analog binding in primary cultures of ovine pituitary cells . Finally , the presence of membrane-associated E2 binding sites on ovine pituitary cells was demonstrated using a fluorescein isothiocyanate conjugate of E2-BSA . We suggest that E2 regulation of GnRHR number during the preovulatory period reflects a membrane site of action and may proceed through a nonclassical signaling mechanism , specifically a CREB-dependent pathway . P12821 inhibition suppresses plasminogen activator inhibitor-1 expression in the neointima of balloon-injured rat aorta . BACKGROUND : P00747 activator inhibitor-1 ( P05121 ) , an important regulator of fibrinolysis and extracellular matrix turnover , has been implicated in a number of vascular diseases . Studies demonstrating angiotensin II ( Ang II ) to be a potent stimulator of P05121 expression in cultured vascular cells suggests that the renin-angiotensin system may modulate vascular P05121 expression . METHODS AND RESULTS : We examined the effects of the P12821 inhibitor captopril on P05121 expression in control and balloon-injured rat aorta . Northern blot analysis demonstrated that aortic P05121 mRNA expression was 7.6-fold elevated 3 hours ( P < .05 ) after balloon injury , back to baseline at 2 days , increased again at 4 days , and by 7 days after balloon injury was 3.2-fold elevated ( P < .05 ) when compared with control . In captopril-treated rats , the induction of P05121 expression by balloon injury was significantly suppressed by 44 % ( P < .05 ) in the 7 day group but was not altered in the 3-hour group . DB01197 also reduced baseline aortic P05121 mRNA . In situ hybridization and immunohistochemistry revealed dense P05121 staining of 7-day neointima in untreated rats and a dramatic decrease in P05121 in neointima of captopril-treated rats . CONCLUSIONS : This report demonstrates that balloon injury results in both a rapid P12821 inhibitor-independent induction of aortic P05121 expression and a later increase in P05121 in the neointima that is significantly suppressed by captopril . This provides the first evidence that the renin-angiotensin system regulates neointimal P05121 expression and that P12821 inhibitors can reduce P05121 in the vessel wall in vivo . DB08890 - a secretagogue and antihyperalgesic agent - what next ? Ongoing clinical trials suggest that linaclotide , a first-in-class , 14-amino acid peptide guanylate cyclase-C ( P25092 ) receptor agonist and intestinal secretagogue is an effective treatment for chronic constipation . A study in this issue of the Journal suggests that linaclotide also has antihyperalgesic effects in three common rat models of inflammation- and stress-induced hypersensitivity ( i.e. , acute trinitrobenzene sulfonic acid colitis , water avoidance stress [ P42768 ] , and restraint-induced stress ) but not in naïve animals . In mice , linaclotide at least partly reduces hyperalgesia via P25092 receptors . Dose-effect relationships of linaclotide were complicated and non-linear . This viewpoint discusses human clinical trials with linaclotide and the results of this study . Potential mechanisms and clinical significance of these findings are explored . Collectively , these data suggest that P25092 receptors exert other , as yet poorly understood , effects on gastrointestinal sensitivity in conditions associated with inflammation and/or stress-induced increased intestinal permeability . However , the data need to be confirmed in humans and in long-term animal models . Further studies are also necessary to elucidate the mechanisms as these effects can not be explained by linaclotide 's known effects on epithelial P25092 receptors . Chemopreventive efficacy and mechanism of licofelone in a mouse lung tumor model via aspiration . Our previous study comparing inhalation and aspiration to administer agents directly to lung indicated that aspiration route is as effective as inhalation while reducing costs for equipment and chemopreventive agent . This study evaluated the chemopreventive efficacy and mechanism of licofelone , a dual inhibitor of P35354 and P09917 ( 5-Lox ) , via oropharyngeal aspiration against mouse lung adenoma . Eight-week-old female A/J mice were given three doses of benzo[a]pyrene ( B[a]P ; 2 mg/dose , gavage ) to induce lung adenomas . After dysplasia developed , the mice were given licofelone ( 0 , 0.03 , 0.1 , or 0.3 mg/kg ) for 16 weeks , and tumor incidence and multiplicity in lung were measured . In addition , the expression of a series of biomarkers in lung cancer progression was evaluated at 2 and 16 weeks . DB04725 showed dose-related inhibition of B[a]P-induced tumor incidence and multiplicity at 0.03 and 0.1 mg/kg following 16-week treatment . DB04725 also showed dose-dependent inhibition of P35354 ( 25 % -41 % ) and 5-Lox ( 35 % -61 % ) at 2 and 16 weeks and proliferating cell nuclear antigen ( P12004 ; 41 % -61 % ) at 16 weeks . A dose-dependent increase in apoptosis ( 1.5- to 2.4-fold ) was also observed in licofelone groups . A marginal inhibition of survivin was observed at one dose . In conclusion , this study showed that licofelone via aspiration showed chemopreventive efficacy against mouse lung adenoma with good correlation to early and late biomarkers of lung cancer progression . This is the first study to show that the aspiration route can be an excellent inexpensive alternative to inhalation for direct delivery of drugs to rodent lungs for efficacy testing of potential chemopreventive agents . Albumin caused the increasing production of angiotensin II due to the dysregulation of P12821 / Q9BYF1 expression in P52789 cells . BACKGROUND : Previous studies have proposed that albuminuria is involved in the activation of intrarenal renin angiotensin system ( DB01367 ) , while its potential mechanism is still unclear . We investigated the influence of albumin on the expression of P12821 / Q9BYF1 and generation of Ang II in P52789 cells . METHODS : The mRNA and protein expression of P12821 and Q9BYF1 was determined by RT-PCR and western blot respectively . Cellular localization of P12821 and Q9BYF1 was shown by laser scanning confocal microscope ( LSCM ) . The concentration of Ang II in the supernatant was detected by radioimmunoassay ( RIA ) . RESULTS : Treatment of HK-2 cells to BSA led to a significant increasing expression of P12821 mRNA in dose and time dependent manner . The overexpression of P12821 protein induced by BSA was consistent with its mRNA expression . Meanwhile , the mRNA and protein expression of Q9BYF1 upon the stimulation of BSA was significantly downregulated in dose and time dependent manner . BSA could significantly increase the production of Ang II in the supernatant ( p < 0.05 ) . DB01197 , however , attenuated the expression of P12821 but increased expression of Q9BYF1 induced by BSA . CONCLUSION : These findings suggested a novel insight on the albuminuria induced activation of intrarenal DB01367 by upregulation of P12821 and downregulation of Q9BYF1 . A review of vilazodone , serotonin , and major depressive disorder . OBJECTIVE : To review the mechanism of selective serotonin reuptake inhibitor ( SSRI ) -mediated serotonergic neurotransmission , focusing on serotonin 1A ( P08908 ) autoreceptors , which are proposed to be involved in delaying therapeutic efficacy . DB06684 was specifically designed to function both as an SSRI and a partial agonist at P08908 receptors . This combined mechanism is proposed to decrease time to efficacy , minimize sexual side effects , and provide concomitant anxiolytic properties . DATA SOURCES : A PubMed search of all English-language articles from January 1990 to January 2013 was conducted using the search terms depression and 5-HT 1A , depression and buspirone , depression and pindolol , and vilazodone . STUDY SELECTION : We found 47 articles and abstracts that were selected for inclusion on the basis of information about the pharmacology of P08908 receptors and the clinical data on pindolol , buspirone , and vilazodone in depression . DATA EXTRACTION : This review summarizes current literature involving antidepressant activity , the role of P08908 autoreceptors , and clinical trials involving serotonin reuptake inhibition in conjunction with P08908 agonists and partial agonists , with a focus on vilazodone . RESULTS : DB06684 has demonstrated efficacy in 2 large , randomized , double-blind , placebo-controlled trials in major depressive disorder . RESULTS suggest that vilazodone has a low incidence of sexual side effects and is effective in patients with high levels of anxiety . A pooled analysis shows evidence of significant symptom reduction after only 1 week of therapy . CONCLUSIONS : If future studies corroborate the clinical benefits attributed to its mechanism of action , vilazodone may show potential advantages in terms of onset of action , sexual side effects , and anxiolytic activity in patients with major depressive disorder . DB08885 for the treatment of neovascular age-related macular degeneration . BACKGROUND : Age-related macular degeneration ( AMD ) affects > 14 million individuals worldwide . Although 90 % of patients with AMD have the dry form , neovascular AMD accounts for the vast majority of patients who develop legal blindness . Until recently , few treatment options existed for treatment of neovascular AMD . The advent of anti- P15692 therapy has significantly improved the safe and effective treatment of neovascular AMD . In addition to two anti- P15692 drugs currently in widespread use , ranibizumab and bevacizumab , a number of medications that interrupt angiogenesis are currently under investigation . One promising new drug is aflibercept ( DB08885 ) , a fusion protein that blocks all isoforms of P15692 and placental growth factors-1 and -2 . OBJECTIVE : To review the current literature and clinical trial data regarding DB08885 for the treatment of neovascular AMD . METHODS : Literature review . RESULTS/CONCLUSION : DB08885 is a novel anti- P15692 therapy , with Phase I and II trial data indicating safety , tolerability and efficacy for the treatment of neovascular AMD . Two Phase III clinical trials ( VIEW-1 and VIEW-2 ) comparing DB08885 to ranibizumab are currently continuing and will provide vital insight into the clinical applicability of this drug . DB08890 -a novel secretagogue in the treatment of irritable bowel syndrome with constipation and chronic idiopathic constipation . Irritable bowel syndrome with constipation ( IBS-C ) and chronic idiopathic constipation ( Q96RK0 ) are highly prevalent gastrointestinal disorders . Traditional symptoms based therapies had somewhat limited success and efficacy in addressing the disorders . Recently , linaclotide emerged as novel peptide capable of improving abdominal symptoms in patients suffering from IBS-C and Q96RK0 . Guanylate cyclase C ( P25092 ) receptor a multi domain protein , found to be molecular target for linaclotide which acts by activating P25092 receptor on the apical surface of intestinal epithelial cells . Binding of linaclotide to P25092 receptor triggers the elevation of second messenger cGMP that elicits fluid secretion into intestinal cells which play a critical role in maintaining homeostasis through cystic fibrosis transmembrane conductance regulator ( P13569 ) . Data from Phase II and III clinical trials demonstrated that linaclotide seems to produce a statistically significant increase in stool frequency , improved straining , decreased abdominal pain and discomfort . Fermented soshiho-tang with Lactobacillus plantarum enhances the antiproliferative activity in vascular smooth muscle cell . BACKGROUND : Soshiho-tang ( P61278 ) is a traditional medicine widely used for the treatment of chronic hepatitis . P61278 has been shown to confer a variety of pharmacological activities , including prevention of hepatotoxicity , promotion of liver regeneration , and modulation of liver fibrosis . In this study , we investigated the antiproliferative activity of native and fermented ( FSST ) formulations of P61278 in vascular smooth muscle cells ( VSMCs ) and examined the potential underlying mechanisms driving these effects . METHODS : P61278 , along with preparations fermented with Lactobacillus plantarum KFRI-144 ( S-A144 ) , L. amylophilus KFRI-161 ( S-A161 ) and L. bulgaricus KFRI-344 ( S-A344 ) , were investigated to determine their effects on the proliferation and viability of VSMCs , along with the signalling pathways underlying these effects . RESULTS : S-A144 exhibited a strong , dose-dependent inhibition of VSMC proliferation relative to untreated controls , but the others did not affect . In addition , S-A144 significantly decreased the phosphorylation of Akt and PLCγ1 in a dose-dependent manner and induced cell cycle arrest at the G0/ P55008 phase characterised by decreased expression of CDKs , cyclins and P12004 . CONCLUSIONS : The findings suggest that S-A144 exhibit enhanced inhibition of DB00102 -induced VSMC proliferation comparison to S-AOR through the suppression of cell cycle progression and expression of cell cycle-related proteins , along with the downregulation of Akt phosphorylation .