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BigScience Biomedical Datasets 112

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Induction of apoptosis of Beta cells of the pancreas by advanced glycation end-products , important mediators of chronic complications of diabetes mellitus . We herein report cytotoxicity of advanced glycation end-products ( AGEs ) on pancreatic beta cells . AGEs stimulated reactive oxygen species ( ROS ) generation but did not arrest proliferation of the P01308 -1 cell line . Pancreatic beta cell lines or primary cultured islets possess a receptor for P51606 ( RAGE ) , and its expression increased after P51606 treatment . TUNEL staining and FACS analysis using annexin V/PI antibodies showed that apoptosis increased in P01308 -1 cells or primary cultured islets when incubated with BSA conjugated with glyceraldehyde ( AGE2 ) or glucoaldehyde ( AGE3 ) , compared with those conjugated with glucose ( AGE1 ) . Reaction of P01308 -1 cells to Ki67 , which is a cellular marker for proliferation , was also increased after P51606 treatment . The ability of primary cultured islets to secrete insulin was retained even after P51606 treatment under either low or high glucose conditions . The antiserum against RAGE partially prevented P51606 -induced cellular events . Treatment of beta cells with the antioxidant metallothionein results in a significant reduction in pathologic changes . AGEs might be able to induce apoptosis as well as proliferation of pancreatic beta cell lines or primary cultured islets . Moreover , antibody array showed that Q06609 and P43351 were significantly decreased in AGE2-treated P01308 -1 cells . AGEs might inhibit homologous DNA recombination for repairing DNA of P01308 -1 cells damaged by ROS generation . It might be suggested that treatment of AGEs resulted in ROS production and apoptosis through their receptor on pancreatic beta cells . AGEs might deteriorate function of pancreatic beta cells in patients with long-term hyperglycemia . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . Potentiation of neurotoxicity in double-mutant mice with Pink1 ablation and A53T- P37840 overexpression . The common age-related neurodegeneration of Parkinson 's disease can result from dominant causes like increased dosage of vesicle-associated alpha-synuclein ( P37840 ) or recessive causes like deficiency of mitophagy factor Q9BXM7 . Interactions between these triggers and their convergence onto shared pathways are crucial , but currently conflicting evidence exists . Here , we crossed previously characterized mice with A53T- P37840 overexpression and with Pink1 deletion to generate double mutants ( DMs ) . We studied their lifespan and behavior , histological and molecular anomalies at late and early ages . DM animals showed potentiated phenotypes in comparison with both single mutants ( SMs ) , with reduced survival and strongly reduced spontaneous movements from the age of 3 months onwards . In contrast to SMs , a quarter of DM animals manifested progressive paralysis at ages > 1 year and exhibited protein aggregates immunopositive for pSer129- P37840 , p62 and ubiquitin in spinal cord and basal brain . Brain proteome quantifications of ubiquitination sites documented altered degradation of P37840 and the DNA-damage marker P16104 at the age of 18 months . Global brain transcriptome profiles and qPCR validation experiments identified many consistent transcriptional dysregulations already at the age of 6 weeks , which were absent from SMs . The observed downregulations for Dapk1 , Dcaf17 , Rab42 and the novel P37840 -marker Lect1 as well as the upregulations for Dctn5 , Mrpl9 , Tmem181a , Xaf1 and H2afx reflect changes in ubiquitination , mitochondrial/synaptic/microtubular/cell adhesion dynamics and DNA damage . Thus , our study confirmed that P37840 -triggered neurotoxicity is exacerbated by the absence of Q9BXM7 and identified a novel molecular signature that is detectable early in the course of this double pathology . P37840 A30P point-mutation generates age-dependent nigrostriatal deficiency in mice . Lewy bodies are mainly composed of alpha-synuclein ( P37840 ) and specific mutations in P37840 gene are related to familial forms of Parkinson 's disease ( PD ) . The purpose of our study was to generate a mouse line with A30P knock-in point mutation in P37840 gene and to test if a single point-mutation is able to turn otherwise normal P37840 into a toxic form . The behavioral profile of P37840 A30P mice was followed for 16 months . Generally , these mice are healthy and viable without any obvious abnormalities . Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function ( ink-test and beam walk ) . In other tests ( motility boxes , rotarod ) mice continuously performed normally . Moreover , P37840 A30P mice expressed the altered sensitivity to Q05940 inhibitor reserpine , possibly reflecting a functional deficiency of dopamine . Indeed , mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum , and reduced levels of dopamine in the mesolimbic system . The present study confirms that P37840 plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance . The generated mouse line has a potential to become a model for PD with comparable time course and phenotype . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Growth-associated gene expression profiles by microarray analysis of trophoblast of molar pregnancies and normal villi . We used microarray analysis to investigate expression profiles of 589 known genes committed to cell growth control to characterize regulatory circuitry for cell proliferation in complete moles ( CMs ) . CMs are characterized by hyperplastic trophoblast and have a high propensity to give rise to choriocarcinoma . Characteristic alterations in gene expression profiles were observed when compared with normal villi . Fifty-seven genes were significantly up-regulated in CMs and involved the Ras-Map kinase 3 , Jak- P42229 , and Wnt signal pathways , implicating growth factor or cytokine-mediated signal pathways in the trophoblastic hyperplasia of CMs . Several genes associated with anti-apoptosis , cell structuring , and/or cell attachment were also up-regulated in CMs . In contrast , relatively fewer genes were down-regulated and these involved IGFBPs , versican , interleukin-1 , tumor necrosis factor receptor , P16070 , and P43351 . Genes identified in this study may elucidate regulation mechanisms of trophoblastic proliferation and mechanisms causing a pathological phenotype in CMs . Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries . BACKGROUND : Atherosclerosis affects aorta , coronary , carotid , and iliac arteries most frequently than any other body vessel . There may be common molecular pathways sustaining this process . Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis . RESULTS : We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression . We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis . Caveolae and JAK/ P35610 pathways , and P06702 / P05109 interacting proteins are certainly involved in the development of vascular disease . We found that the system of caveolae is directly connected with genes that respond to hormone receptors , and indirectly with the apoptosis pathway . Cytokines , chemokines and growth factors released in the blood flux were investigated in parallel . High levels of RANTES , IL-1ra , MIP-1 alpha , MIP-1 beta , P60568 , P05112 , P05113 , P05231 , P13232 , Q16552 , DB00102 , P15692 and P01579 were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels . CONCLUSION : The pattern of cytokine and P06702 / P05109 up-regulation characterizes atherosclerosis as a proinflammatory disorder . Activation of the JAK/ P35610 pathway is confirmed by the up-regulation of P05231 , P42224 , Q00978 and Q13651 genes in coronary and carotid plaques . The functional network constructed in our research is an evidence of the central role of P35610 protein and the caveolae system to contribute to preserve the plaque . Moreover , Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Molecular determinants of trastuzumab efficacy : What is their clinical relevance ? DB00072 -containing therapy is a standard of care for human epidermal growth factor receptor-2 ( P04626 ) -positive breast cancer . In pre-clinical models , a wide range of molecular mechanisms have been associated with reduced sensitivity to trastuzumab in vitro . These include expression of the truncated P04626 receptor fragment p95HER2 , activating mutation of the gene encoding the class 1A catalytic subunit of phosphatidylinositol 3-kinase ( P42336 ) , loss of phosphatase and tensin homolog ( P60484 ) , activation of other downstream signal transducers , prevention of cell cycle arrest , increased signaling through alternative ( HER or non-HER ) tyrosine kinase receptors , and resistance to antibody-dependent cellular cytotoxicity . However , the clinical significance of these mechanisms as determinants of trastuzumab efficacy in vivo has been unclear . Here , we review clinical studies of potential predictive biomarkers of trastuzumab efficacy in P04626 -positive breast cancer and consider whether evaluation of such markers might inform patient selection for therapy . We find that clinical evidence relating to potential predictive biomarkers is mostly limited to small , retrospective studies , many of which have yielded conflicting findings . Some trends are evident in the retrospective data and in biomarker analyses from randomized clinical trials , particularly relating to activation of the phosphatidylinositol 3-kinase pathway , but none is sufficiently strong to form a basis for patient selection . This may be explained by the fact that multiple mechanisms of action determine the clinical efficacy of trastuzumab . In the absence of novel , validated biomarkers of efficacy , trastuzumab eligibility should continue to be based on evaluation of P04626 status according to standard methods . Recombinant P17936 inhibits allergic lung inflammation , P15692 production , and vascular leak in a mouse model of asthma . BACKGROUND : Vascular endothelial growth factor ( P15692 ) plays a pro-inflammatory mediator as well as a vascular permeability factor in bronchial asthma . P01308 -like growth factor ( IGF ) -I is also involved in the inflammatory process associated with bronchial asthma and stimulates P15692 expression . The IGF-binding proteins ( IGFBPs ) , especially P17936 , display distinctive properties and can interfere with various biological processes . METHODS : In this study , an ovalbumin ( OVA ) -induced murine model of allergic airway disease was used to investigate which mechanism is implicated in the preventive and therapeutic actions of P17936 administered exogenously on allergen-induced bronchial inflammation and airway hyper-responsiveness , in particular focusing on the regulation of P15692 expression . RESULTS : Administration of recombinant human P17936 to OVA-inhaled mice substantially attenuated the increases in hypoxia-inducible factor ( HIF ) -α activity , P05019 production , and P15692 protein levels in the lung . In addition , the blockade of P05019 action decreased the OVA-induced P15692 expression , airway inflammation , and bronchial hyper-responsiveness . The administration of recombinant human P17936 or CBO-P11 also reduced significantly increases in inflammatory cells , airway hyper-responsiveness , levels of P05112 , P05113 , P35225 , and vascular permeability in the lung of OVA-inhaled mice . Moreover , when recombinant human P17936 was administered after the completion of OVA inhalation , these therapeutic effects of P17936 were also observed . CONCLUSIONS : These results indicate that P17936 administered exogenously may attenuate antigen-induced airway inflammation and hyper-responsiveness through the modulation of vascular leakage and P15692 expression mediated by HIF-1α/HIF-2α signaling as well as P05019 action in allergic airway disease of mice . P10275 accelerates premature senescence of human dermal papilla cells in association with DNA damage . The dermal papilla , located in the hair follicle , expresses androgen receptor and plays an important role in hair growth . Androgen/ P10275 actions have been implicated in the pathogenesis of androgenetic alopecia , but the exact mechanism is not well known . Recent studies suggest that balding dermal papilla cells exhibit premature senescence , upregulation of p16(INK4a) , and nuclear expression of DNA damage markers . To investigate whether androgen/AR signaling influences the premature senescence of dermal papilla cells , we first compared frontal scalp dermal papilla cells of androgenetic alopecia patients with matched normal controls and observed that premature senescence is more prominent in the dermal papilla cells of androgenetic alopecia patients . Exposure of androgen induced premature senescence in dermal papilla cells from non-balding frontal and transitional zone of balding scalp follicles but not in beard follicles . Overexpression of the AR promoted androgen-induced premature senescence in association with p16(INK4a) upregulation , whereas knockdown of the androgen receptor diminished the effects of androgen . An analysis of γ- P16104 expression in response to androgen/androgen receptor signaling suggested that DNA damage contributes to androgen/androgen receptor-accelerated premature senescence . These results define androgen/androgen receptor signaling as an accelerator of premature senescence in dermal papilla cells and suggest that the androgen/androgen receptor-mediated DNA damage-p16(INK4a) axis is a potential therapeutic target in the treatment of androgenetic alopecia . DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . Antenatal maternally-administered phosphodiesterase type 5 inhibitors normalize P29474 expression in the fetal lamb model of congenital diaphragmatic hernia . PURPOSE : Pulmonary hypertension ( pHTN ) , a main determinant of survival in congenital diaphragmatic hernia ( Q8NE62 ) , results from in utero vascular remodeling . Phosphodiesterase type 5 ( O76074 ) inhibitors have never been used antenatally to treat pHTN . The purpose of this study is to determine if antenatal O76074 inhibitors can prevent pHTN in the fetal lamb model of Q8NE62 . METHODS : Q8NE62 was created in pregnant ewes . Postoperatively , pregnant ewes received oral placebo or tadalafil , a O76074 inhibitor , until delivery . Near term gestation , lambs underwent resuscitations , and lung tissue was snap frozen for protein analysis . RESULTS : Mean cGMP levels were 0.53±0.11 in placebo-treated fetal lambs and 1.73±0.21 in tadalafil-treated fetal lambs ( p=0.002 ) . Normalized expression of P29474 was 82 % ±12 % in Normal-Placebo , 61 % ±5 % in Q8NE62 -Placebo , 116 % ±6 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . Normalized expression of β-sGC was 105 % ±15 % in Normal-Placebo , 82 % ±3 % in Q8NE62 -Placebo , 158 % ±16 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . P29474 and β-sGC were significantly decreased in Q8NE62 ( p=0.0007 and 0.01 for P29474 and β-sGC , respectively ) , and tadalafil significantly increased P29474 expression ( p=0.0002 ) . CONCLUSIONS : O76074 inhibitors can cross the placental barrier . β-sGC and P29474 are downregulated in fetal lambs with Q8NE62 . Antenatal O76074 inhibitors normalize P29474 and may prevent in utero vascular remodeling in Q8NE62 . Regulation of double-strand break-induced mammalian homologous recombination by P63165 , a Q96B01 . Mammalian Q06609 protein plays essential roles in DNA homologous recombination , DNA repair and cell proliferation . Q06609 activities are regulated by its associated proteins . It was previously reported that a ubiquitin-like protein , P63165 , associates with Q06609 in the yeast two-hybrid system . One function of P63165 is to covalently conjugate with target proteins and thus modify their function . In the present study we found that non-conjugated P63165 forms a complex with Q06609 and P43351 proteins in human cells . Overexpression of P63165 down-regulates DNA double-strand break-induced homologous recombination in CHO cells and reduces cellular resistance to ionizing radiation in HT1080 cells . With or without overexpressed P63165 , most homologous recombination products arise by gene conversion . However , overexpression of P63165 reduces the fraction of bidirectional gene conversion tracts . Overexpression of a mutant P63165 that is incapable of being conjugated retains the ability to inhibit homologous recombination . These results suggest a regulatory role for P63165 in homologous recombination . DB00184 induces cell proliferation , invasion and epithelial-mesenchymal transition in a variety of human cancer cell lines . Cigarette smoking is strongly correlated with the onset of nonsmall cell lung cancer ( NSCLC ) . DB00184 , an active component of cigarettes , has been found to induce proliferation of lung cancer cell lines . In addition , nicotine can induce angiogenesis and confer resistance to apoptosis . All these events are mediated through the nicotinic acetylcholine receptors ( nAChRs ) on lung cancer cells . In this study , we demonstrate that nicotine can promote anchorage-independent growth in NSCLCs . In addition , nicotine also induces morphological changes characteristic of a migratory , invasive phenotype in NSCLCs on collagen gel . These morphological changes were similar to those induced by the promigratory growth factor P15692 . The proinvasive effects of nicotine were mediated by alpha7-nAChRs on NSCLCs . RT-PCR analysis showed that the alpha7-nAChRs were also expressed on human breast cancer and pancreatic cancer cell lines . DB00184 was found to promote proliferation and invasion in human breast cancer . The proinvasive effects of nicotine were mediated via a nAChR , Src and calcium-dependent signaling pathway in breast cancer cells . In a similar fashion , nicotine could also induce proliferation and invasion of Aspc1 pancreatic cancer cells . Most importantly , nicotine could induce changes in gene expression consistent with epithelial to mesenchymal transition ( EMT ) , characterized by reduction of epithelial markers like P12830 expression , ZO-1 staining and concomitant increase in levels of mesenchymal proteins like vimentin and fibronectin in human breast and lung cancer cells . Therefore , it is probable that the ability of nicotine to induce invasion and EMT may contribute to the progression of breast and lung cancers . Topoisomerase II-mediated DNA cleavage and mutagenesis activated by nitric oxide underlie the inflammation-associated tumorigenesis . AIMS : Both cancer-suppressing and cancer-promoting properties of reactive nitrogen and oxygen species ( RNOS ) have been suggested to play a role in tumor pathology , particularly those activities associated with chronic inflammation . Here , we address the impact of nitric oxide ( NO ) on the induction of DNA damage and genome instability with a specific focus on the involvement of topoisomerase II ( P11388 ) . We also investigate the contribution of NO to the formation of skin melanoma in mice . RESULTS : Similar to the P11388 -targeting drug , etoposide ( DB00773 ) , the NO-donor , S-nitrosoglutathione ( GSNO ) , induces skin melanomas formation in 7,12-dimethyl- benz[a]anthracene ( DMBA ) -initiated mice . To explore the mechanism(s) underlying this NO-induced tumorigenesis , we use a co-culture model system to demonstrate that inflamed macrophages with inducible NO synthase ( P35228 ) expression cause γ- P16104 activation , p53 phosphorylation , and chromosome DNA breaks in the target cells . Inhibitor experiments revealed that NO and P11388 isozymes are responsible for the above described cellular phenotypes . Notably , NO , unlike DB00773 , preferentially induces the formation of TOP2β cleavable complexes ( TOP2βcc ) in cells . Moreover , GSNO induced P11388 -dependent DNA sequence rearrangements and cytotoxicity . Furthermore , the incidences of GSNO- and DB00773 -induced skin melanomas were also observed to be lower in the skin-specific top2β-knockout mice . Our results suggest that P11388 isozymes contribute to NO-induced mutagenesis and subsequent cancer development during chronic inflammation . INNOVATION AND CONCLUSIONS : We provide the first experimental evidence for the functional role of P11388 in NO-caused DNA damage , mutagenesis , and carcinogenesis . Notably , these studies contribute to our molecular understanding of the cancer-promoting actions of RNOS during chronic inflammation . Changing paradigms in management of metastatic Castration Resistant Prostate Cancer ( mCRPC ) . Recently , the standard of care for metastatic Castration Resistant Prostate Cancer ( mCRPC ) has changed considerably . Persistent androgen receptor ( AR ) signaling has been identified as a target for novel therapies and reengages the fact that AR continues to be the primary target responsible for metastatic prostate cancer . P10275 gene amplification and over expression have been found to result in a higher concentration of androgen receptors on tumor cells , making them extremely sensitive to low levels of circulating androgens . Additionally , prostate cancer cells are able to maintain dihydrotestosterone ( DB02901 ) concentration in excess of serum concentrations to support tumor growth . For many years ketoconazole was the only P05093 inhibitor that was used to treat mCRPC . However , significant toxicities limit its use . Newly approved chemotherapeutic agents such as DB05812 ( an oral selective inhibitor of CYP17A ) , which blocks androgen biosynthesis both within and outside the prostate cancer cells ) , and enzalutamide ( blocks AR signaling ) have improved overall survival . There are also ongoing phase III trials for Orteronel ( P50750 - 700 ) , ARN- 509 and Galeterone ( TOK-001 ) , which targets androgen signaling . In this review , we will present the rationale for the newly approved hormonal treatments , their indications and complications , and we will discuss ongoing trials that are being done to improve the efficacy of the approved agents . Finally , we will talk about the potential upcoming hormonal treatments for mCRPC . Sporadic breast carcinomas with somatic P38398 gene deletions share genotype/phenotype features with familial breast carcinomas . BACKGROUND : High frequencies of loss of heterozygosity ( LOH ) are found in familial breast carcinomas with BRCA mutations . Although LOH of P38398 does not coincide with somatic P38398 mutations , reduced P38398 protein expression and hypermethylation indicate the involvement of P38398 in sporadic carcinogenesis . To further investigate the role of BRCA we determined LOH of P38398 and correlated this with LOH in other breast cancer-associated regions . MATERIALS AND METHODS : A total of 105 sporadic breast carcinomas were analysed for LOH in the regions of P38398 , P51587 , P04637 , Caveolin1 , " putative BRCA3 " , P60484 , Q13315 and P12830 and correlated it with clinicopathological features . RESULTS : We found an overall increase of LOH in carcinomas with simultaneous LOH of P38398 . Significantly higher LOH rates were detected in the regions of P04637 ( 80 % : 34.7 % ; p < 0.005 ) , 8q21 ( 72.7 % : 30.6 % ; p < 0.010 ) and 10q22-23 ( 21.1 % : 5.9 % ; p=0.043 ) . Moreover , estrogen receptor-negative carcinomas revealed LOH of P38398 more frequently than estrogen receptor-positive carcinomas ( 39 % : 12 % ; p=0.003 ) . CONCLUSION : These data indicate that LOH of P38398 coincides with a defect of the DNA repair pathway . Therefore , LOH of P38398 determines a subgroup of sporadic breast carcinomas sharing genotype/phenotype features with familial breast carcinomas . Q00987 is a ubiquitin ligase of P12956 -Akt promotes cell survival by inhibiting Q00987 -dependent P12956 destabilization . Earlier , we have reported that 70 kDa subunit of Ku protein heterodimer ( P12956 ) binds and inhibits Bax activity in the cytosol and that ubiquitin ( Ub ) -dependent proteolysis of cytosolic P12956 facilitates Bax-mediated apoptosis . We found that Q00987 ( human homolog of murine double minute ) has an ability to ubiquitinate P12956 and that Q00987 overexpression in cultured cells causes a decrease in P12956 expression levels . An interaction between P12956 and Q00987 was shown by means of immunoprecipitation , whereas none could be shown between 80 kDa subunit of Ku protein heterodimer and Q00987 . Vascular endothelial growth factor ( P15692 ) is known to inhibit endothelial cell ( EC ) apoptosis through an Akt-mediated survival kinase signal ; however , the mechanism underlying this inhibition of apoptosis has not been fully elucidated . We found that P15692 inhibited cytosolic P12956 degradation induced by apoptotic stress . It is known that Akt-dependent phosphorylation of Q00987 causes nuclear translocation of Q00987 followed by Q00987 -mediated inactivation of p53 . We found that P15692 stimulated nuclear translocation of Q00987 in EC and efficiently inhibited P12956 degradation . We also found that constitutively active Akt , but not kinase-dead Akt , inhibited P12956 degradation in the cytosol . Furthermore , P12956 knockdown diminished antiapoptotic activity of Akt . Taken together , we propose that Q00987 is a P12956 Ub ligase and that Akt inhibits Bax-mediated apoptosis , at least in part , by maintaining P12956 levels through the promotion of Q00987 nuclear translocation . Loss of homologous recombination or non-homologous end-joining leads to radial formation following DNA interstrand crosslink damage . High levels of interstrand cross-link damage in mammalian cells cause chromatid breaks and radial formations recognizable by cytogenetic examination . The mechanism of radial formation observed following DNA damage has yet to be determined . Due to recent findings linking homologous recombination and non-homologous end-joining to the action of the Fanconi anemia pathway , we speculated that radials might be the result of defects in either of the pathways of DNA repair . To test this hypothesis , we have investigated the role of homologous recombination proteins Q06609 and P43351 , non-homologous end-joining proteins P12956 and P49917 , and protein P49959 in radial formation and cell survival following interstrand crosslink damage with mitomycin C . For the studies we used small inhibitory RNA to deplete the proteins from cells , allowing for evaluation of radial formation and cell survival . In transformed normal human fibroblasts , depletion of these proteins increased interstrand crosslink sensitivity as manifested by decreased cell survival and increased radial formation . These results demonstrate that inactivation of proteins from either of the two separate DNA repair pathways increases cellular sensitivity to interstrand crosslinks , indicating each pathway plays a role in the normal response to interstrand crosslink damage . We can also conclude that homologous recombination or non-homologous end-joining are not required for radial formation , since radials occur with depletion of these pathways . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Mitoxantrone inhibits HIF-1α expression in a topoisomerase II-independent pathway . PURPOSE : Solid tumors encounter a growth-limiting hypoxic microenvironment as they develop . Hypoxia-inducible factors ( HIF ) play important roles in hypoxia-associated tumor development and therapeutic resistance . Targeting the HIF pathway ( especially HIF-1α ) represents a promising cancer treatment strategy . Here , we report a novel class of HIF-1α inhibitors and the possible molecular basis of inhibition . EXPERIMENTAL DESIGN : We analyzed the inhibitory effects of clinically used topoisomerase II ( P11388 ) -targeting drugs on HIF-1α expression with a primary focus on mitoxantrone . The potential role of P11388 in mitoxantrone-inhibited HIF-1α expression was studied using pharmacologic inhibition , a knockdown approach , and P11388 mutant cells . Moreover , involvement of mitoxantrone in proteasome-mediated degradation , transcription , and translation of HIF-1α was examined . RESULTS : The P11388 -targeting mitoxantrone , but neither doxorubicin nor etoposide ( DB00773 ) , strongly inhibited HIF-1α expression under hypoxic conditions in a dose- and time-dependent manner . Surprisingly , the mitoxantrone-mediated inhibition of HIF-1α expression was largely independent of two P11388 isozymes , proteasomal degradation , and transcription . Furthermore , mitoxantrone inhibited HIF-1α expression and function in a similar fashion as cycloheximide , suggesting that mitoxantrone might inhibit HIF-1α via a blockage at its translation step . In vitro translation experiments using HIF-1α mRNA further confirmed inhibition of HIF-1α translation by mitoxantrone . Interestingly , levels of the polysome-bound HIF-1α and P15692 mRNA were elevated and decreased after mitoxantrone treatment , respectively . CONCLUSIONS : We have identified the P11388 -targeting compound , mitoxantrone , as an HIF-1α inhibitor possibly through a translation inhibition mechanism , suggesting the possibility of an additional anticancer activity for mitoxantrone . Blockade of cannabinoid receptors reduces inflammation , leukocyte accumulation and neovascularization in a model of sponge-induced inflammatory angiogenesis . OBJECTIVE : Angiogenesis depends on a complex interaction between cellular networks and mediators . The endocannabinoid system and its receptors have been shown to play a role in models of inflammation . Here , we investigated whether blockade of cannabinoid receptors may interfere with inflammatory angiogenesis . MATERIALS AND METHODS : Polyester-polyurethane sponges were implanted in C57Bl/6j mice . Animals received doses ( 3 and 10 mg/kg/daily , s.c. ) of the cannabinoid receptor antagonists SR141716A ( P21554 ) or SR144528 ( CB2 ) . Implants were collected at days 7 and 14 for cytokines , hemoglobin , myeloperoxidase , and N-acetylglucosaminidase measurements , as indices of inflammation , angiogenesis , neutrophil and macrophage accumulation , respectively . Histological and morphometric analysis were also performed . RESULTS : Cannabinoid receptors expression in implants was detected from day 4 after implantation . Treatment with P21554 or CB2 receptor antagonists reduced cellular influx into sponges at days 7 and 14 after implantation , although P21554 receptor antagonist were more effective at blocking leukocyte accumulation . There was a reduction in P01375 -α , P15692 , P09341 /KC , P13500 /JE , and P10147 /MIP-1α levels , with increase in P13501 /RANTES . Both treatments reduced neovascularization . Dual blockade of cannabinoid receptors resulted in maximum inhibition of inflammatory angiogenesis . CONCLUSIONS : Blockade of cannabinoid receptors reduced leukocyte accumulation , inflammation and neovascularization , suggesting an important role of endocannabinoids in sponge-induced inflammatory angiogenesis both via P21554 and CB2 receptors . DB00072 has opposing effects on SN-38-induced double-strand breaks and cytotoxicity in P04626 -positive gastric cancer cells depending on administration sequence . AIM : We investigated the effects of trastuzumab , an anti- P04626 humanized monoclonal antibody , on DNA breaks induced by SN-38 , a topoisomerase-1 inhibitor , in gastric cancer cell lines positive or negative for P04626 expression . MATERIALS AND METHODS : NCI-N87 ( P04626 + ) and MKN74 ( P04626 - ) cells were exposed to SN-38 in the presence or absence of trastuzumab . DB00072 was added either prior to or after SN-38 . Effects of trastuzumab on the induction of gamma- P16104 , a marker of DNA double-strand breaks , the cytotoxicity of SN-38 and cell cycle progression were determined . RESULTS : When trastuzumab was administered following SN-38 , it increased γ P16104 levels and cytotoxicity of SN-38 in NCI-N87 cells , but not in MKN74 cells . In contrast , pretreatment with trastuzumab reduced SN-38-induced γ P16104 expression and cytotoxicity of SN-38 in NCI-N87 cells , but not in MKN74 cells . DB00072 delayed cell cycle progression in NCI-N87 cells only . CONCLUSION : DB00072 has opposing effects on SN-38-induced double-strand breaks and cytotoxicity depending on the order of administration of the two agents . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Guggulsterone inhibits angiogenesis by blocking P40763 and P15692 expression in colon cancer cells . The plant sterol guggulsterone has been shown to exert anti-tumor effects , making it a candidate chemotherapeutic agent . We investigated the anti-tumor effects of guggulsterone on colon cancer cells and elucidated the underlying molecular mechanisms related to angiogenesis . The apoptotic effects of guggulsterone were examined by cell survival assay . Western blot analysis was used to determine the levels of various down-stream intracellular proteins involved in angiogenesis , including signal transducer and activator of transcription 3 ( P40763 ) , vascular endothelial growth factor ( P15692 ) , hypoxia-inducible factor-1alpha ( HIF-1alpha ) and aryl hydrocarbon receptor nuclear translocator ( P27540 ) . Using chromatin immunoprecipitation assay , we tested whether guggulsterone affects the recruitment of P40763 , P27540 and HIF-1alpha to the human P15692 promoter . To investigate the effect of guggulsterone on vascular endothelial cell migration and invasion , tube formation and migration assays were conducted using human umbilical vein endothelial cells ( HUVECs ) . Matrix metalloproteinase ( MMP ) -2 and -9 activities were measured by gelatin zymography . Guggulsterone significantly reduced cell viability in colon cancer cells in a dose-dependent manner and blocked P15692 , P27540 and P40763 expression prominently in hypoxic conditions . The recruitment of P40763 and P27540 , but not HIF-1alpha , to the P15692 promoter was inhibited by guggulsterone treatment . HUVECs produced much foreshortened and severely broken tubes and showed decreased migration activity under guggulsterone effects . In addition , zymography revealed that P08253 and -9 enzyme activities were markedly lower in the presence of guggulsterone . The results of this study suggest that guggulsterone not only induces apoptosis , but also inhibits angiogenesis and metastasis in colon cancer cells by blocking P40763 and P15692 expression , suggesting its therapeutic potential in the treatment of colorectal cancer . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . Role of nitrative and oxidative DNA damage in inflammation-related carcinogenesis . Chronic inflammation induced by biological , chemical , and physical factors has been found to be associated with the increased risk of cancer in various organs . We revealed that infectious agents including liver fluke , Helicobacter pylori , and human papilloma virus and noninfectious agents such as asbestos fiber induced P35228 -dependent formation of 8-nitroguanine and 8-oxo-7 , 8-dihydro-2'-deoxyguanosine ( 8-oxodG ) in cancer tissues and precancerous regions . Our results with the colocalization of phosphorylated Q13315 and γ- P16104 with 8-oxodG and 8-nitroguanine in inflammation-related cancer tissues suggest that DNA base damage leads to double-stranded breaks . It is interesting from the aspect of genetic instability . We also demonstrated P05231 -modulated P35228 expression via P40763 and P00533 in Epstein-Barr-virus-associated nasopharyngeal carcinoma and found promoter hypermethylation in several tumor suppressor genes . Such epigenetic alteration may occur by controlling the DNA methylation through P05231 -mediated JAK/ P40763 pathways . Collectively , 8-nitroguanine would be a useful biomarker for predicting the risk of inflammation-related cancers . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside ameliorates vascular senescence and improves blood flow involving a mechanism of p53 deacetylation . BACKGROUND AND AIMS : 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside ( THSG ) , a resveratrol analog with glucoside , has been shown in various studies to inhibit proliferation of vascular smooth muscle cells , attenuate inflammation , and prevent vascular endothelial dysfunction . In the study , we examined the effects of THSG on vascular senescence and blood flow . METHODS AND RESULTS : Oral administration of THSG for 14 weeks , resulted in notable increases in blood flow in spontaneously hypertensive rats ( SHRs ) ; and effective inhibition of vascular senescence as indicated by senescence-associated β-galactosidase ( SA-β-gal ) staining , phosphorylation of γ P16104 observed by stain analysis of immunofluorescence , and K373 acetylation of p53 in the aortic arches of SHRs . Oral administration of THSG also induced P29474 expression and urinary NOx production . THSG weekly activated Q96EB6 activity , stimulated P29474 promoter reporter gene activity , and ameliorated H(2)O(2)-induced cellular senescence and K373 acetylation of p53 in cultured human umbilical vein endothelial cells ( HUVECs ) . CONCLUSIONS : THSG improves blood flow and ameliorates vascular senescence by increasing P29474 expression and Sirt1 activity and decreasing acetylation of p53 at K373 site , at least in part , both in vitro and in vivo . Amsacrine and etoposide hypersensitivity of yeast cells overexpressing DNA topoisomerase II . Increasing the cellular concentration of DNA topoisomerase II in yeast by expressing constitutively a plasmid-borne P11388 gene encoding the enzyme greatly increases the sensitivity of the cells to amsacrine and etoposide ( DB00773 ) . This increased drug sensitivity at a higher intracellular DNA topoisomerase II level is observed in both P43351 + repair-proficient strains and rad52 mutants that are defective in the repair of double-stranded breaks . These results provide strong support of the hypothesis that the cellular target of these drugs is DNA topoisomerase II , and that these drugs kill cells by converting DNA topoisomerase II into a DNA damaging agent . Program death-1 engagement upon TCR activation has distinct effects on costimulation and cytokine-driven proliferation : attenuation of Q9Y6W8 , P05112 , and Q9HBE4 , but not P10747 , P13232 , and P40933 responses . The program death 1 ( P18621 ) receptor and its ligands , P18621 ligand (PD-L)1 and Q9BQ51 , define a novel regulatory pathway with potential inhibitory effects on T , B , and monocyte responses . In the present study , we show that human P01730 (+) T cells express P18621 , Q9NZQ7 , and Q9BQ51 upon activation , and Abs to the receptor can be agonists or antagonists of the pathway . Under optimal conditions of stimulation , Q9Y6W8 but not P10747 costimulation can be prevented by P18621 engagement . P60568 levels induced by costimulation are critical in determining the outcome of the P18621 engagement . Thus , low to marginal P60568 levels produced upon Q9Y6W8 costimulation account for the greater sensitivity of this pathway to P18621 -mediated inhibition . Interestingly , exogenous P60568 , P13232 , and P40933 but not P05112 and Q9HBE4 can rescue P18621 inhibition , suggesting that among these cytokines only those that activate P42229 can rescue P18621 inhibition . As P42229 has been implicated in the maintenance of IL-2Ralpha expression , these results suggest that P13232 and P40933 restore proliferation under conditions of P18621 engagement by enhancing high-affinity IL-2R expression and hence , P60568 responsiveness . Regulation of microphthalmia-associated transcription factor O75030 protein levels by association with the ubiquitin-conjugating enzyme hUBC9 . The basic helix-loop-helix/leucine zipper ( bHLH/ Q8N5A5 ) microphthalmia-associated transcription factor ( O75030 ) regulates transcription of genes encoding enzymes essential for melanin biosynthesis in melanocytes and retinal pigmented epithelial cells . To determine how O75030 activity is regulated , we used the yeast two-hybrid system to identify proteins expressed by human melanoma cells that interact with O75030 . The majority of clones that showed positive interaction with a 158-amino-acid region of O75030 containing the bHLH/ Q8N5A5 domain ( aa 168-325 ) encoded the ubiquitin conjugating enzyme hUBC9 . The association of O75030 with hUBC9 was further confirmed by an in vitro Q86UG4 pull-down assay . Although hUBC9 is known to interact preferentially with SENTRIN/ P63165 , in vitro transcription/translation analysis demonstrated greater association of O75030 with ubiquitin than with SENTRIN . Importantly , cotransfection of O75030 and hUBC9 expression vectors resulted in O75030 protein degradation . O75030 protein was stabilized by the proteasome inhibitor MG132 , indicating the role of the ubiquitin-proteasome system in O75030 degradation . DB00133 73 , which is located in a region rich in proline , glutamic acid , serine , and threonine ( PEST ) , regulates O75030 protein stability , since a serine to alanine mutation prevented hUBC9-mediated O75030 ( S73A ) degradation . Furthermore , we identified lysine 201 as a potential ubiquitination site . A lysine to arginine mutation abolished O75030 ( K201R ) degradation by hUBC9 in vivo . Our experiments indicate that by targeting O75030 for proteasome degradation , hUBC9 is a critical regulator of melanocyte differentiation . P60484 sumo-wrestles human P43351 to mystery land . DB00588 -induced regulation of the balance within macrophage subpopulations . In asthma , treatment with inhaled corticosteroids reduces chronic peribronchial inflammation and restores the balance within macrophage subpopulations . This study investigates whether corticosteroids can regulate monocyte differentiation in vitro and thereby influence the balance of functionally distinct macrophages . Graded doses of fluticasone propionate ( FP ) were added to cultures of normal peripheral blood monocytes in the presence or absence of P05112 . Cells were harvested after 7 days ' culture . Double immunofluorescence studies were performed on cytospins of differentiated macrophages using the MoAbs RFD1 and RFD7 to distinguish inductive and suppressive macrophages by their respective phenotypes . Macrophage function was determined by quantifying allostimulation in a mixed leucocyte reaction and by measuring tumour necrosis factor-alpha ( P01375 ) production . FP reduced the number of mature cells with a D1+ antigen-presenting phenotype and up-regulated the development of cells with the D1/D7+ and D7+ phenotypes . Functionally , this was associated with reduced stimulation of T cell proliferation in a mixed leucocyte reaction ( P08235 ) . DB00588 also reversed the increase in both D1+ expression and P01375 production induced by P05112 . The effect of FP persisted for 24 h after removal of FP from the culture medium . These results suggest that FP treatment of asthmatics may have a direct beneficial effect by normalizing the macrophage subset imbalance that contributes to the chronic peribronchial inflammation present in this condition . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Selective inhibition of histone deacetylase 6 ( Q9UBN7 ) induces DNA damage and sensitizes transformed cells to anticancer agents . Q9UBN7 ( Q9UBN7 ) is structurally and functionally unique among the 11 human zinc-dependent histone deacetylases . Here we show that chemical inhibition with the Q9UBN7 -selective inhibitor tubacin significantly enhances cell death induced by the topoisomerase II inhibitors etoposide and doxorubicin and the pan-HDAC inhibitor DB02546 ( vorinostat ) in transformed cells ( LNCaP , MCF-7 ) , an effect not observed in normal cells ( human foreskin fibroblast cells ) . The inactive analogue of tubacin , nil-tubacin , does not sensitize transformed cells to these anticancer agents . Further , we show that down-regulation of Q9UBN7 expression by shRNA in LNCaP cells enhances cell death induced by etoposide , doxorubicin , and DB02546 . Tubacin in combination with DB02546 or etoposide is more potent than either drug alone in activating the intrinsic apoptotic pathway in transformed cells , as evidenced by an increase in PARP cleavage and partial inhibition of this effect by the pan-caspase inhibitor Z-VAD-fmk . Q9UBN7 inhibition with tubacin induces the accumulation of γ P16104 , an early marker of DNA double-strand breaks . Tubacin enhances DNA damage induced by etoposide or DB02546 as indicated by increased accumulation of γ P16104 and activation of the checkpoint kinase Chk2 . Tubacin induces the expression of P35638 ( P35638 / P35638 ) , a transcription factor up-regulated in response to cellular stress . P35638 induction is further increased when tubacin is combined with DB02546 . These findings point to mechanisms by which Q9UBN7 -selective inhibition can enhance the efficacy of certain anti-cancer agents in transformed cells . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . Current researches on breast cancer epidemiology in Korea . As a cause of death in women , breast cancer ranks second to stomach cancer in Korea . Age-standardized mortality rates for breast cancer steadily increased during the 1980s and 1990s . There are big differences in the incidence rates for breast cancer compared with Western countries . Epidemiological features , trends in morbidity and mortality , various age-specific incidence curves , migrant study results , and analysis of the risk factors , however , suggest that the incidence of breast cancer might be further increasing in Korea . The key epidemiological hormonal risk factors for breast cancer are all explicable in terms of the estrogen augmented by progesterone hypothesis . These include older age , family history of breast cancer , early menarche , late menopause , late full-term pregnancy , and never a breast feeding . Both the establishment of high-risk groups and the estimation of lifetime risk are essential to develop a control strategy against breast cancer . Invasive ductal carcinoma is the most common histologic type of breast cancer in Korea , and the five-year survival rate has been estimated as 80-83 % . Recent studies on the identification of susceptibility factors such as genetic polymorphisms of P09488 /T1/P1 , P21964 , P05181 , P11511 , P05093 , P03372 , P18887 , O43542 , P43351 , TGF-alpha , P01375 , IL-1B , IL-1RN , P50613 etc. that predispose individuals to breast cancer by gene-environment or gene-gene interactions may possibly give further insight into both the etiology and the prevention of this malignancy . Acidic pH induces topoisomerase II-mediated DNA damage . Acidic pH plays an important role in various pathophysiological states and has been demonstrated to be carcinogenic in animal models . Recent studies have also implicated acidic pH in the development of preneoplastic Barrett 's esophagus in human . However , little is known about the molecular mechanism underlying acidic pH-induced carcinogenesis . In the current study , we show that acidic pH , like the topoisomerase II ( P11388 ) poison DB00773 ( demethylepipodophyllotoxin ethylidene-beta-D-glucoside ) , induces tumors in 9,10-dimethyl-1,2-benzanthracene(DMBA)-initiated mice . The following studies in tissue culture models have suggested that acidic pH acts like a P11388 poison to induce P11388 -mediated DNA damage : ( i ) acidic pH induces P11388 -dependent DNA damage signals as evidenced by up-regulation of p53 and DB00133 -139 phosphorylation of P16104 [ a substrate for ataxia telangiectasia mutated ( Q13315 ) Q13315 and Rad3-related ( ATR ) kinases ] ; ( ii ) acidic pH-induced cytotoxicity in tumor cells is reduced in P11388 -deficient cells ; ( iii ) acidic pH increases the mutation frequency of the hypoxanthine phosphoribosyl transferase ( P00492 ) gene in a P11388 -dependent manner ; and ( iv ) acidic pH induces reversible P11388 -mediated DNA strand breaks in vitro . We discuss the possibility that P11388 -mediated DNA damage may contribute to acidic pH-induced carcinogenesis . Senescence-associated secretory phenotype in a mouse model of bleomycin-induced lung injury . DB00290 produces DNA damage , apoptosis and senescence , all of which play crucial roles in the development of pulmonary fibrosis . Recently , close attention has been paid to a DNA damage-induced phenotypic change ( senescence-associated secretory phenotype ; SASP ) as a trigger for the secretion of various mediators which modify the processes of tissue injury , inflammation , repair and fibrosis . We characterized the SASP in a murine model of bleomycin-induced lung injury . Mice were intratracheally administered bleomycin or control saline , and the lungs were obtained on days 7 , 14 and 21 . The occurrence of DNA damage and the SASP in the lungs was examined by immunostaining . γ P16104 immunostaining of the bleomycin-treated lungs revealed double-strand breaks ( DSBs ) , largely within P12830 -positive , β4-integirn-positive alveolar epithelial cells . The DSBs were associated with phosphorylation of Q13315 /ATR , a central signal transducer mediating the DNA damage response , and upregulation of the cyclin-dependent kinase inhibitor P38936 (CIP1) . The DSBs persisted for at least 21 days after the bleomycin exposure , although it began to wane after 7 days . A subpopulation of the γ P16104 -positive , DNA-damaged cells exhibited the SASP , characterized by overexpression of P05231 , TNFα , P08253 and P14780 , in association with the phosphorylation of IKKα/β and p38 MAPK . Persistent DNA damage and the SASP are induced in the process of bleomycin-induced lung injury and repair , suggesting that these events play an important role in the regulation of inflammation and tissue remodeling in bleomycin-induced pneumopathy . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . DB00820 , a further innovation in the treatment of sexual dysfunction . In recognition of the large number of sufferers of sexual dysfunction worldwide , and the variety of etiologies of the condition , investigation into effective pharmacological agents has been expanded . One method of intervention is inhibition of the phosphodiesterase type 5 ( O76074 ) enzyme , which has already been exploited with a considerable degree -- though not complete -- success . A number of new agents that inhibit O76074 are under development . Notable among these is tadalafil , which has demonstrated a high level of selectivity for O76074 over the other phosphodiesterases and has shown efficacy in improving erectile function and sexual satisfaction in phase III trials . Throughout the clinical development program for tadalafil , the drug has been well tolerated and without serious side effects . The manufacturer , Lilly Q9Y6W8 , received an approvable letter from the US Food and Drug Administration for use of the drug as a treatment for erectile dysfunction on April 30 , 2002 . Lilly Q9Y6W8 hopes to market tadalafil , with the trade name DB00820 , in the USA in 2003 . A p53 axis regulates B cell receptor-triggered , innate immune system-driven B cell clonal expansion . Resting mature human B cells undergo a dynamic process of clonal expansion , followed by clonal contraction , during an in vitro response to surrogate C3d-coated Ag and innate immune system cytokines , P05112 and Q9Y275 . In this study , we explore the mechanism for clonal contraction through following the time- and division-influenced expression of several pro- and anti-apoptotic proteins within CFSE-labeled cultures . Several findings , involving both human and mouse B cells , show that a mitochondria-dependent apoptotic pathway involving p53 contributes to the high activation-induced cell death ( AICD ) susceptibility of replicating blasts . Activated B cell clones exhibit elevated p53 protein and elevated mRNA/protein of proapoptotic molecules known to be under direct p53 transcriptional control , Bax , Bad , Puma , Bid , and procaspase 6 , accompanied by reduced anti-apoptotic Bcl-2 . Under these conditions , Bim levels were not increased . The finding that full-length Bid protein significantly declines in AICD-susceptible replicating blasts , whereas Bid mRNA does not , suggests that Bid is actively cleaved to short-lived , proapoptotic truncated Bid . AICD was diminished , albeit not eliminated , by p53 small interfering RNA transfection , genetic deletion of p53 , or Bcl-2 overexpression . DNA damage is a likely trigger for p53-dependent AICD because susceptible lymphoblasts expressed significantly elevated levels of both phosphorylated ataxia telangiectasia mutated- DB00133 (1980) and phospho- P16104 - DB00133 (139) . Deficiency in activation-induced cytosine deaminase diminishes but does not ablate murine B cell AICD , indicating that activation-induced cytosine deaminase-induced DNA damage is only in part responsible . Evidence for p53-influenced AICD during this route of T cell-independent clonal expansion raises the possibility that progeny bearing p53 mutations might undergo positive selection in peripherally inflamed tissues with elevated levels of P05112 and Q9Y275 . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development .

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interacts_with DB09079?

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Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries . BACKGROUND : Atherosclerosis affects aorta , coronary , carotid , and iliac arteries most frequently than any other body vessel . There may be common molecular pathways sustaining this process . Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis . RESULTS : We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression . We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis . Caveolae and JAK/ P35610 pathways , and P06702 / P05109 interacting proteins are certainly involved in the development of vascular disease . We found that the system of caveolae is directly connected with genes that respond to hormone receptors , and indirectly with the apoptosis pathway . Cytokines , chemokines and growth factors released in the blood flux were investigated in parallel . High levels of RANTES , IL-1ra , MIP-1 alpha , MIP-1 beta , P60568 , P05112 , P05113 , P05231 , P13232 , Q16552 , DB00102 , P15692 and P01579 were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels . CONCLUSION : The pattern of cytokine and P06702 / P05109 up-regulation characterizes atherosclerosis as a proinflammatory disorder . Activation of the JAK/ P35610 pathway is confirmed by the up-regulation of P05231 , P42224 , Q00978 and Q13651 genes in coronary and carotid plaques . The functional network constructed in our research is an evidence of the central role of P35610 protein and the caveolae system to contribute to preserve the plaque . Moreover , Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . DB00184 induces cell proliferation , invasion and epithelial-mesenchymal transition in a variety of human cancer cell lines . Cigarette smoking is strongly correlated with the onset of nonsmall cell lung cancer ( NSCLC ) . DB00184 , an active component of cigarettes , has been found to induce proliferation of lung cancer cell lines . In addition , nicotine can induce angiogenesis and confer resistance to apoptosis . All these events are mediated through the nicotinic acetylcholine receptors ( nAChRs ) on lung cancer cells . In this study , we demonstrate that nicotine can promote anchorage-independent growth in NSCLCs . In addition , nicotine also induces morphological changes characteristic of a migratory , invasive phenotype in NSCLCs on collagen gel . These morphological changes were similar to those induced by the promigratory growth factor P15692 . The proinvasive effects of nicotine were mediated by alpha7-nAChRs on NSCLCs . RT-PCR analysis showed that the alpha7-nAChRs were also expressed on human breast cancer and pancreatic cancer cell lines . DB00184 was found to promote proliferation and invasion in human breast cancer . The proinvasive effects of nicotine were mediated via a nAChR , Src and calcium-dependent signaling pathway in breast cancer cells . In a similar fashion , nicotine could also induce proliferation and invasion of Aspc1 pancreatic cancer cells . Most importantly , nicotine could induce changes in gene expression consistent with epithelial to mesenchymal transition ( EMT ) , characterized by reduction of epithelial markers like P12830 expression , ZO-1 staining and concomitant increase in levels of mesenchymal proteins like vimentin and fibronectin in human breast and lung cancer cells . Therefore , it is probable that the ability of nicotine to induce invasion and EMT may contribute to the progression of breast and lung cancers . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Modeling the neurovascular niche : murine strain differences mimic the range of responses to chronic hypoxia in the premature newborn . Preterm birth results in significant cognitive and motor disabilities , but recent evidence suggests that there is variable recovery over time . One possibility that may explain this variable recovery entails variable neurogenic responses in the subventricular zone ( SVZ ) following the period of chronic hypoxia experienced by these neonates . In this report , we have characterized the responses to chronic hypoxia of two mouse strains that represent a wide range of susceptibility to chronic hypoxia . We determined that C57BL/6 pups and neural progenitor cells ( NPCs ) derived from them exhibit a blunted response to hypoxic insult compared with CD-1 pups and NPCs . Specifically , C57BL/6 pups and NPCs exhibited blunted in vivo and in vitro proliferative and increased apoptotic responses to hypoxic insult . Additionally , C57BL/6 NPCs exhibited lower baseline levels and hypoxia-induced levels of selected transcription factors , growth factors , and receptors ( including HIF-1alpha , Q9GZT9 , P23560 , P15692 , P48061 , TrkB , Nrp-1 , P61073 , and NO ) that determine , in part , the responsiveness to chronic hypoxic insult compared with CD-1 pups and NPCs , providing insight into this important and timely problem in perinatology . 4-[3,5-Bis(trimethylsilyl)benzamido] benzoic acid inhibits angiogenesis in colon cancer through reduced expression of vascular endothelial growth factor . 4-[3,5-bis(trimethylsilyl)benzamido] Benzoic acid ( TAC-101 ) has potent antiproliferative , antiangiogenic , and antitumor effects in vitro and in vivo . These effects might be due to TAC-101 binding to retinoic acid receptor alpha ( P10276 ) and interfering with the binding of activator protein-1 ( AP-1 ) to DNA . However , little is known about the detailed mechanism of TAC-101 function . We investigated the mechanism of the antiangiogenic effect of TAC-101 using a rat hepatic metastatic model in vivo and DLD-1 human colon cancer cells in vitro . Liver metastases were induced by portal injection of Q15293 -9 rat colonic cancer cells into F344 rats . TAC-101 ( 8 mg/kg ) was orally administered 5 days per week for 4 weeks and then hepatic tumors were immunohistochemically evaluated for microvessel density ( P53602 ) and vascular endothelial growth factor ( P15692 ) . TAC-101 significantly reduced both P53602 and P15692 expression . Northern blot analysis and ELISA indicated that TAC-101 efficiently inhibited production of P15692 mRNA and protein in DLD-1 cells in a time- and dose-dependent manner . These findings suggest that TAC-101 may inhibit progression and metastasis in colon cancer by interfering with tumor production of P15692 . 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 . DB09079 , a triple angiokinase inhibitor , enhances cytotoxic therapy response in pancreatic cancer . Angiogenesis remains a sensible target for pancreatic ductal adenocarcinoma ( PDAC ) therapy . P15692 , PDGF , FGF and their receptors are expressed at high levels and correlate with poor prognosis in human PDAC . DB09079 is a triple angiokinase inhibitor that targets P17948 /2/3 , P11362 /2/3 and PDGFRα/β signaling . We investigated the antitumor activity of nintedanib alone or in combination with the cytotoxic agent gemcitabine in experimental PDAC . DB09079 inhibited proliferation of cells from multiple lineages found in PDAC , with gemcitabine enhancing inhibitory effects . DB09079 blocked PI3K/MAPK activity and induced apoptosis in vitro and in vivo . In a heterotopic model , net local tumor growth compared to controls ( 100 % ) was 60.8 ± 10.5 % in the gemcitabine group , -2.1 ± 9.9 % after nintedanib therapy and -12.4 ± 16 % after gemcitabine plus nintedanib therapy . Effects of therapy on intratumoral proliferation , microvessel density and apoptosis corresponded with tumor growth inhibition data . In a PDAC survival model , median animal survival after gemcitabine , nintedanib and gemcitabine plus nintedanib was 25 , 31 and 38 days , respectively , compared to 16 days in controls . The strong antitumor activity of nintedanib in experimental PDAC supports the potential of nintedanib-controlled mechanisms as targets for improved clinical PDAC therapy . Q00987 is a ubiquitin ligase of P12956 -Akt promotes cell survival by inhibiting Q00987 -dependent P12956 destabilization . Earlier , we have reported that 70 kDa subunit of Ku protein heterodimer ( P12956 ) binds and inhibits Bax activity in the cytosol and that ubiquitin ( Ub ) -dependent proteolysis of cytosolic P12956 facilitates Bax-mediated apoptosis . We found that Q00987 ( human homolog of murine double minute ) has an ability to ubiquitinate P12956 and that Q00987 overexpression in cultured cells causes a decrease in P12956 expression levels . An interaction between P12956 and Q00987 was shown by means of immunoprecipitation , whereas none could be shown between 80 kDa subunit of Ku protein heterodimer and Q00987 . Vascular endothelial growth factor ( P15692 ) is known to inhibit endothelial cell ( EC ) apoptosis through an Akt-mediated survival kinase signal ; however , the mechanism underlying this inhibition of apoptosis has not been fully elucidated . We found that P15692 inhibited cytosolic P12956 degradation induced by apoptotic stress . It is known that Akt-dependent phosphorylation of Q00987 causes nuclear translocation of Q00987 followed by Q00987 -mediated inactivation of p53 . We found that P15692 stimulated nuclear translocation of Q00987 in EC and efficiently inhibited P12956 degradation . We also found that constitutively active Akt , but not kinase-dead Akt , inhibited P12956 degradation in the cytosol . Furthermore , P12956 knockdown diminished antiapoptotic activity of Akt . Taken together , we propose that Q00987 is a P12956 Ub ligase and that Akt inhibits Bax-mediated apoptosis , at least in part , by maintaining P12956 levels through the promotion of Q00987 nuclear translocation . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Loss of epigenetic Kruppel-like factor 4 histone deacetylase ( KLF-4-HDAC ) -mediated transcriptional suppression is crucial in increasing vascular endothelial growth factor ( P15692 ) expression in breast cancer . Vascular endothelial growth factor ( P15692 ) is recognized as an important angiogenic factor that promotes angiogenesis in a series of pathological conditions , including cancer , inflammation , and ischemic disorders . We have recently shown that the inflammatory transcription factor P56270 is , at least in part , responsible for the marked increase of P15692 levels in breast cancer . Here , we show that P56270 -mediated induction of P15692 is repressed by KLF-4 transcription factor . KLF-4 is abundantly present in normal breast epithelial cells , but its level is considerably reduced in breast cancer cells and clinical cancer tissues . In the human P15692 promoter , P56270 - and KLF-4-binding elements are overlapping , whereas P56270 induces and KLF-4 suppresses P15692 expression . Ectopic overexpression of KLF-4 and RNAi-mediated inhibition of endogenous KLF-4 supported the role of KLF-4 as a transcriptional repressor of P15692 and an inhibitor of angiogenesis in breast cancer cells . We show that KLF-4 recruits histone deacetylases ( HDACs ) -2 and -3 at the P15692 promoter . Chronological ChIP assays demonstrated the occupancy of KLF-4 , Q92769 , and O15379 in the P15692 promoter in normal MCF-10A cells but not in MDA-MB-231 cancer cells . Co-transfection of KLF-4 and HDAC expression plasmids in breast cancer cells results in synergistic repression of P15692 expression and inhibition of angiogenic potential of these carcinoma cells . Together these results identify a new mechanism of P15692 up-regulation in cancer that involves concomitant loss of KLF-4-HDAC-mediated transcriptional repression and active recruitment of P56270 -mediated transcriptional activation . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . 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 .

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interacts_with DB00083?

multiple_choice

Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . Synthesis of substrates and inhibitors of botulinum neurotoxin type A metalloprotease . Botulinum neurotoxin ( BoNT ) metalloproteases and related proteases are the most selective proteases known . X-ray crystal structures suggest that the active sites of the native enzymes exist in catalytically incompetent forms that must be activated by substrate binding . In order to characterize the postulated substrate-induced conformational changes for enzyme activation , we synthesized a series of transition-state analog inhibitors in which the dipeptide cleavage site is replaced by tetrahedral intermediate analogs within the minimal substrate peptide sequence . In this paper , we report our efforts to design inhibitors of DB00083 metalloprotease . We confirm that an effective substrate sequence for DB00083 metalloprotease is a 17-mer peptide corresponding to residues 187-203 of P60880 . A more stable substrate , Nle202SNAP-25 [ 187-203 ] was synthesized in order to develop an assay for proteolytic activity of DB00083 metalloprotease that can be used to monitor time-dependent inhibition . Alpha-thiol amide analogs of Gln-197 were incorporated via solid-phase peptide synthesis into both 17-mer minimal peptide substrate sequences . The synthesis , characterization and inhibition kinetics for the alpha-thiol amide analogs of holotoxin A substrate are described . These substrate-derived inhibitors were shown to be submicromolar inhibitors of DB00083 catalytic activity . Comparison of the catalytic properties of the botulinum neurotoxin subtypes A1 and A5 . Clostridium botulinum neurotoxins ( BoNTs ) cause the life-threatening disease botulism through the inhibition of neurotransmitter release by cleaving essential SNARE proteins . There are seven serologically distinctive types of BoNTs and many subtypes within a serotype have been identified . BoNT/A5 is a recently discovered subtype of type A botulinum neurotoxin which possesses a very high degree of sequence similarity and identity to the well-studied A1 subtype . In the present study , we examined the endopeptidase activity of these two DB00083 subtypes and our results revealed significant differences in substrate binding and cleavage efficiency between subtype A5 and A1 . Distinctive hydrolysis efficiency was observed between the two toxins during cleavage of the native substrate P60880 versus a shortened peptide mimic . N-terminal truncation studies demonstrated that a key region of the P60880 , including the amino acid residues at 151 through 154 located in the remote binding region of the substrate , contributed to the differential catalytic properties between A1 and A5 . Elevated binding affinity of the peptide substrate resulted from including these important residues and enhanced BoNT/A5 's hydrolysis efficiency . In addition , mutations of these amino acid residues affect the proteolytic performance of the two toxins in different ways . This study provides a better understanding of the biological activity of these toxins , their performance characteristics in the Endopep-MS assay to detect BoNT in clinical samples and foods , and is useful for the development of peptide substrates . Comparative role of neurotoxin-associated proteins in the structural stability and endopeptidase activity of botulinum neurotoxin complex types A and E . Seven serotypes of botulinum neurotoxins , the most toxic substances known to mankind , are each produced by different strains of Clostridium botulinum along with a group of neurotoxin-associated proteins ( NAPs ) . NAPs play a critical role in the toxicoinfection process of botulism in addition to their role in protecting the neurotoxin from proteolytic digestion in the GI tract as well as from adverse environmental conditions . In this study we have investigated the effect of temperature on the structural and functional stability of DB00083 complex ( BoNT/AC ) and BoNT/E complex ( BoNT/EC ) . Although the NAPs in the two complexes are quite different , both groups of NAPs activate the endopeptidase activities of their BoNTs without any need to reduce the disulfide bonds between light and heavy chains of respective BoNTs . BoNT/AC attains optimum enzyme activity at the physiological temperature of 37 degrees C whereas BoNT/EC is maximally active at 45 degrees C , and this is accompanied by conformational alterations in its polypeptide folding at this temperature , leading to favorable binding with its intracellular substrate , P60880 , and subsequent cleavage of the latter . DB00083 in its complex form is found to be structurally more stable against temperature whereas BoNT/E in its complex form is functionally better protected against temperature . Based on the analysis of isolated NAPs we have observed that the structural stability of the BoNT/AC is contributed by the NAPs . In addition to the unique structural conditions in which the enzyme remains active , functional stability of botulinum neurotoxins against temperature plays a critical role in the survival of the agent in cooked food and in food-borne botulism . New insights into clostridial neurotoxin-SNARE interactions . Botulinum neurotoxin serotype A ( DB00083 ) has achieved a dichotomous status in modern medicine ; it is both a versatile treatment for several neurological disorders and a lethal poison responsible for causing the neuroparalytic syndrome botulism . The extent of paralysis largely depends on the dosage of toxin received . The toxins block neurotransmitter release by delivering their DB01593 (2+)-dependent protease components to the presynaptic side of chemical synapses . These highly specialized enzymes exclusively hydrolyze peptide bonds within SNARE ( soluble N-ethylmaleiamide-sensitive factor attachment protein receptor ) proteins . Recently , the structural basis for the highly specific interaction between DB00083 and its target SNARE , P60880 ( synaptosomal-associated protein of 25kDa ) , was elucidated . New details regarding the nature of the toxin-SNARE interactions could be exploited for novel inhibitor design . Second generation steroidal 4-aminoquinolines are potent , dual-target inhibitors of the botulinum neurotoxin serotype A metalloprotease and P. falciparum malaria . Significantly more potent second generation 4-amino-7-chloroquinoline ( 4,7-ACQ ) based inhibitors of the botulinum neurotoxin serotype A ( DB00083 ) light chain were synthesized . Introducing an amino group at the C(3) position of the cholate component markedly increased potency ( IC50 values for such derivatives ranged from 0.81 to 2.27 μM ) . Two additional subclasses were prepared : bis(steroidal)-4,7-ACQ derivatives and bis(4,7-ACQ)cholate derivatives ; both classes provided inhibitors with nanomolar-range potencies ( e.g. , the Ki of compound 67 is 0.10 μM ) . During DB00083 challenge using primary neurons , select derivatives protected P60880 by up to 89 % . Docking simulations were performed to rationalize the compounds ' in vitro potencies . In addition to specific residue contacts , coordination of the enzyme 's catalytic zinc and expulsion of the enzyme 's catalytic water were a consistent theme . With respect to antimalarial activity , the compounds provided better IC90 activities against chloroquine resistant ( CQR ) malaria than CQ , and seven compounds were more active than mefloquine against CQR strain W2 . P01375 -alpha induces apoptosis via inducible nitric oxide synthase in neonatal mouse cardiomyocytes . OBJECTIVE : It has been demonstrated that tumor necrosis factor-alpha ( P01375 alpha ) induces apoptosis in cardiac myocytes . However , its mechanism of action is still not well understood . In the present study , we hypothesized that P01375 alpha induces myocardial apoptosis by induction of inducible nitric oxide synthase ( P35228 ) . METHODS : Neonatal cardiac myocytes were isolated from P35228 ( -/- ) mutant and C57BL6 wild type mice . Cells were cultured for 3 days before treatment with an NO donor or P01375 alpha . Following treatment with S-nitroso-N-acetyl-penicillamine ( P60880 ) or P01375 , cells were tested for apoptosis by terminal deoxynucleotidyl transfer-mediated end labeling ( TUNEL ) staining and cell death detection ELISA . NO production was measured by nitrite concentration in the culture medium . Cardiomyocyte expression of P35228 and P01375 type 1 receptor ( P19438 ) mRNA was determined by reverse transcriptase-polymerase chain reaction ( RT-PCR ) . RESULTS : P60880 ( 0.01-100 microM ) induced apoptosis of cardiac myocytes in a concentration-dependent manner in the wild type mice ( n = 5 , P < 0.01 ) . P19438 mRNA was expressed in neonatal cardiomyocytes from both wild type and P35228 ( -/- ) mutant mice . P01375 alpha induced a concentration-dependent increase in P35228 mRNA expression and nitrite production as well as significant apoptosis of cardiomyocytes in the wild type mice ( n = 4 , P < 0.01 ) . However , without P35228 expression , the apoptotic effects of P01375 were significantly attenuated in cardiomyocytes from P35228 ( -/- ) mutant mice ( n = 4 , P < 0.05 ) . CONCLUSION : P01375 alpha induces apoptosis via P35228 expression and NO production in neonatal mouse cardiomyocytes . Inhibition of neurotransmitter release by clostridial neurotoxins correlates with specific proteolysis of synaptosomal proteins . Rat brain synaptosomes were used to study the effect of several clostridial neurotoxins on the neurotransmitter release . In this system the blockade of transmitter release correlated with the proteolytic activity of the toxins . Blockade of glutamate release was linked to selective proteolysis of one of the following synaptic proteins : synaptobrevin ( BoNT/D , BoNT/F ) ; P60880 ( DB00083 , BoNT/E ) , or HPC-1/syntaxin ( BoNT/C1 ) . All the toxins used had an inhibitory effect on synaptosomes with the exception of BoNT/F . BoNT/F cleaved synaptobrevin in permeabilized synaptosomes but failed to produce the same effect on intact synaptosomes . Structural and biochemical studies of botulinum neurotoxin serotype C1 light chain protease : implications for dual substrate specificity . Clostridial neurotoxins are the causative agents of the neuroparalytic disease botulism and tetanus . They block neurotransmitter release through specific proteolysis of one of the three soluble N-ethylmaleimide-sensitive-factor attachment protein receptors ( SNAREs ) P60880 , syntaxin , and synaptobrevin , which constitute part of the synaptic vesicle fusion machinery . The catalytic component of the clostridial neurotoxins is their light chain ( LC ) , a DB01593 endopeptidase . There are seven structurally and functionally related botulinum neurotoxins ( BoNTs ) , termed serotype A to G , and tetanus neurotoxin ( TeNT ) . Each of them exhibits unique specificity for their target SNAREs and peptide bond(s) they cleave . The mechanisms of action for substrate recognition and target cleavage are largely unknown . Here , we report structural and biochemical studies of BoNT/C1-LC , which is unique among BoNTs in that it exhibits dual specificity toward both syntaxin and P60880 . A distinct pocket ( S1 ' ) near the active site likely achieves the correct register for the cleavage site by only allowing Ala as the P1 ' residue for both P60880 and syntaxin . Mutations of this P60880 residue dramatically reduce enzymatic activity . The remote alpha-exosite that was previously identified in the complex of DB00083 -LC and P60880 is structurally conserved in BoNT/C1 . However , mutagenesis experiments show that the alpha-exosite of BoNT/C1 plays a less stringent role in substrate discrimination in comparison to that of DB00083 , which could account for its dual substrate specificity . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Botulinum neurotoxin A activity is dependent upon the presence of specific gangliosides in neuroblastoma cells expressing synaptotagmin I . Botulinum neurotoxin A ( DB00083 ) is the deadliest of all known biological substances . Although its toxicity makes DB00083 a biological warfare threat , its biologic activity makes it an increasingly useful therapeutic agent for the treatment of muscular disorders . However , almost 200 years after its discovery , the neuronal cell components required for the activity of this deadly toxin have not been unequivocally identified . In this work , neuroblastoma cells expressing synaptotagmin I , a protein shown to be bound by DB00083 , were used to determine whether specific gangliosides were necessary for DB00083 activity as measured by synaptosomal-associated protein of 25 kDa ( P60880 ) cleavage . Ganglioside GT1b was found to support DB00083 activity significantly more effectively than GD1a , which was far more effective than GM1 when added to ganglioside-deficient murine cholinergic Neuro 2a or to human adrenergic SK-N-SH neuroblastoma cells . Whereas both cell lines expressed synaptotagmin I , P60880 cleavage was not observed in the absence of complex gangliosides . These results indicate that 1 ) gangliosides are required for DB00083 activity , 2 ) synaptotagmin I in the absence of gangliosides does not support DB00083 activity , and 3 ) Neuro 2a cells are an efficient model system for studying the biological activity of DB00083 . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Substrate recognition mechanism of VAMP/synaptobrevin-cleaving clostridial neurotoxins . Botulinum neurotoxins ( BoNTs ) and tetanus neurotoxin ( TeNT ) inhibit neurotransmitter release by proteolyzing a single peptide bond in one of the three soluble N-ethylmaleimide-sensitive factor attachment protein receptors P60880 , syntaxin , and vesicle-associated membrane protein ( VAMP ) /synaptobrevin . TeNT and BoNT/B , D , F , and G of the seven known BoNTs cleave the synaptic vesicle protein VAMP/synaptobrevin . Except for BoNT/B and TeNT , they cleave unique peptide bonds , and prior work suggested that different substrate segments are required for the interaction of each toxin . Although the mode of P60880 cleavage by DB00083 and E has recently been studied in detail , the mechanism of VAMP/synaptobrevin proteolysis is fragmentary . Here , we report the determination of all substrate residues that are involved in the interaction with BoNT/B , D , and F and TeNT by means of systematic mutagenesis of VAMP/synaptobrevin . For each of the toxins , three or more residues clustered at an N-terminal site remote from the respective scissile bond are identified that affect solely substrate binding . These exosites exhibit different sizes and distances to the scissile peptide bonds for each neurotoxin . Substrate segments C-terminal of the cleavage site ( P4-P4 ' ) do not play a role in the catalytic process . Mutation of residues in the proximity of the scissile bond exclusively affects the turnover number ; however , the importance of individual positions at the cleavage sites varied for each toxin . The data show that , similar to the P60880 proteolyzing DB00083 and E , VAMP/synaptobrevin-specific clostridial neurotoxins also initiate substrate interaction , employing an exosite located N-terminal of the scissile peptide bond . Cytotoxicity of botulinum neurotoxins reveals a direct role of syntaxin 1 and P60880 in neuron survival . Botulinum neurotoxins ( DB00083 -G ) act by blocking synaptic vesicle exocytosis . Whether BoNTs disrupt additional neuronal functions has not been addressed . Here we report that cleavage of syntaxin 1 by BoNT/C , and cleavage of P60880 by BoNT/E both induce degeneration of neurons . Furthermore , although P60880 cleaved by DB00083 still supports neuron survival , it has reduced capacity to tolerate additional mutations . We demonstrate that syntaxin 1 and P60880 cooperate as SNARE proteins to support neuron survival . Exogenous expression of other homologous SNARE proteins , syntaxin 2/3/4 and O00161 , which are resistant to BoNT/C and E in neurons , can substitute syntaxin 1/ P60880 and prevent toxin-induced neuron death . Finally , we find that neuronal death is due to blockage of plasma membrane recycling processes that utilize syntaxin 1/ P60880 , independent of synaptic vesicle exocytosis . These findings establish neuronal cytotoxicity for BoNTs and reveal syntaxin 1/ P60880 as the ubiquitous and essential SNARE proteins mediating multiple fusion events on neuronal plasma membranes . Self-assembled peptide monolayers as a toxin sensing mechanism within arrayed microchannels . A sensor for the lethal bacterial enzyme , botulinum neurotoxin type A ( DB00083 ) , was developed using self-assembled monolayers ( SAMs ) . SAMs consisting of an immobilized synthetic peptide that mimicked the toxin 's in vivo P60880 protein substrate were formed on Au and interfaced with arrayed microfluidic channels . Efforts to optimize DB00118 composition and assay conditions for greatest reaction efficiency and sensitivity are described in detail . Channel design provided facile fluid manipulation , sample incubation , analyte concentration , and fluorescence detection all within a single microfluidic channel , thus avoiding sample transfer and loss . Peptide SAMs were exposed to varying concentrations of DB00083 or its catalytic light chain ( ALC ) , resulting in enzymatic cleavage of the peptide substrate from the surface . Fluorescence detection was achieved down to 20 pg/mL ALC and 3 pg/mL DB00083 in 3 h . Toxin sensing was also accomplished in vegetable soup , demonstrating practicality of the method . The modular design of this microfluidic DB00118 platform allows for extension to sensing other toxins that operate via enzymatic cleavage , such as the remaining BoNT serotypes B-G , anthrax , and tetanus toxin . Accelerated intoxication of GABAergic synapses by botulinum neurotoxin A disinhibits stem cell-derived neuron networks prior to network silencing . Botulinum neurotoxins ( BoNTs ) are extremely potent toxins that specifically cleave SNARE proteins in peripheral synapses , preventing neurotransmitter release . Neuronal responses to BoNT intoxication are traditionally studied by quantifying SNARE protein cleavage in vitro or monitoring physiological paralysis in vivo . Consequently , the dynamic effects of intoxication on synaptic behaviors are not well-understood . We have reported that mouse embryonic stem cell-derived neurons ( ESNs ) are highly sensitive to BoNT based on molecular readouts of intoxication . Here we study the time-dependent changes in synapse- and network-level behaviors following addition of DB00083 to spontaneously active networks of glutamatergic and GABAergic ESNs . Whole-cell patch-clamp recordings indicated that DB00083 rapidly blocked synaptic neurotransmission , confirming that ESNs replicate the functional pathophysiology responsible for clinical botulism . Quantitation of spontaneous neurotransmission in pharmacologically isolated synapses revealed accelerated silencing of GABAergic synapses compared to glutamatergic synapses , which was consistent with the selective accumulation of cleaved P60880 at Q99259 (+) pre-synaptic terminals at early timepoints . Different latencies of intoxication resulted in complex network responses to DB00083 addition , involving rapid disinhibition of stochastic firing followed by network silencing . Synaptic activity was found to be highly sensitive to P60880 cleavage , reflecting the functional consequences of the localized cleavage of the small subpopulation of P60880 that is engaged in neurotransmitter release in the nerve terminal . Collectively these findings illustrate that use of synaptic function assays in networked neurons cultures offers a novel and highly sensitive approach for mechanistic studies of toxin:neuron interactions and synaptic responses to BoNT . Crystal structure of Clostridium botulinum neurotoxin protease in a product-bound state : Evidence for noncanonical zinc protease activity . Clostridium botulinum neurotoxins ( BoNTs ) , the most potent toxins known , disrupt neurotransmission through proteolysis of proteins involved in neuroexocytosis . The light chains of BoNTs are unique zinc proteases that have stringent substrate specificity and require exceptionally long substrates . We have determined the crystal structure of the protease domain from BoNT serotype A ( DB00083 ) . The structure reveals a homodimer in a product-bound state , with loop F242-V257 from each monomer deeply buried in its partner 's catalytic site . The loop , which acts as a substrate , is oriented in reverse of the canonical direction for other zinc proteases . The Y249-Y250 peptide bond of the substrate loop is hydrolyzed , leaving the Y249 product carboxylate coordinated to the catalytic zinc . From the crystal structure of the DB00083 protease , detailed models of noncanonical binding and proteolysis can be derived which we propose are also consistent with DB00083 binding and proteolysis of natural substrate synaptosome-associated protein of 25 kDa ( P60880 ) . The proposed DB00083 substrate-binding mode and catalytic mechanism are markedly different from those previously proposed for the BoNT serotype B . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Longer-acting and highly potent chimaeric inhibitors of excessive exocytosis created with domains from botulinum neurotoxin A and B . Various human neurogenic hyper-excitability disorders are successfully treated with type A or B BoNT ( botulinum neurotoxin ) . The DB00083 complex is widely used because of its longer-lasting benefits ; also , autonomic side-effects are more often reported for BoNT/B . To establish if this distinct effect of BoNT/B could be exploited therapeutically , DB00083 was modified so that it would bind the more abundant BoNT/B acceptor in rodents while retaining its desirable persistent action . The advantageous protease and translocation domain of DB00083 were recombinantly combined with the acceptor-binding moiety of type B [ H(C)/B ( C-terminal half of BoNT/B heavy chain ) ] , creating the chimaera AB . This purified protein bound the BoNT/B acceptor , displayed enhanced capability relative to type A for intraneuronally delivering its protease , cleaved P60880 ( synaptosome-associated protein of 25 kDa ) and induced a more prolonged neuromuscular paralysis than DB00083 in mice . The BA chimaera , generated by substituting H(C)/A ( C-terminal half of DB00083 heavy chain ) into BoNT/B , exhibited an extremely high specific activity , delivered the BoNT/B protease via the DB00083 acceptor into neurons , or fibroblast-like synoviocytes that lack P60880 , cleaving the requisite isoforms of VAMP ( vesicle-associated membrane protein ) . Both chimaeras inhibited neurotransmission in murine bladder smooth muscle . BA has the unique ability to reduce exocytosis from non-neuronal cells expressing the DB00083 -acceptor and utilising VAMP , but not P60880 , in exocytosis . Acute and chronic effects of botulinum neurotoxin a on the mammalian neuromuscular junction . INTRODUCTION : Botulinum neurotoxin A ( DB00083 ) cleaves P60880 and inhibits acetylcholine ( ACh ) release at the neuromuscular junctions ( NMJ ) to cause neuroparalysis . Previous reports indicate a dyssynchrony between the inhibitory effect of DB00083 on ACh release and P60880 cleavage . METHODS : We tested the in vitro ( acute ; 90 min ) and in vivo ( chronic ; 12 h ) effects of DB00083 on stimulus-evoked ACh release ( SEAR ) , twitch tension , and P60880 cleavage in isolated extensor digitorum longus ( Q9Y5X9 ) nerve-muscle preparations ( NMP ) . RESULTS : In vitro or in vivo DB00083 poisoning inhibited SEAR and twitch tension . Conversely , P60880 cleavage and inhibition of spontaneous release frequency were observed only in NMP poisoned with DB00083 in vivo . Moreover , chronic treatment of DB00083 inhibited ionomycin stimulated Ca(2+) signals in Neuro 2a cells . CONCLUSIONS : These results demonstrate that the inhibition of SEAR precedes P60880 cleavage and suggest involvement of a more complex mechanism for the inhibitory effect of DB00083 at the NMJ . Dynamics of motor nerve terminal remodeling unveiled using SNARE-cleaving botulinum toxins : the extent and duration are dictated by the sites of P60880 truncation . Nerve sprouts emerge from motor nerve terminals following blockade of exo-endocytosis for more than 3 days by botulinum neurotoxin ( BoNT ) , and form functional synapses , albeit temporary . Upon restoration of synaptic activity to the parent terminal 7 and 90 days after exposure to BoNT/F or A respectively , a concomitant retraction of the outgrowths was observed . BoNT/E caused short-term neuroparalysis , and dramatically accelerated the recovery of DB00083 -paralyzed muscle by further truncation of P60880 and its replenishment with functional full-length SNARE . The removal of 9 C-terminal residues from P60880 by DB00083 leads to persistence of the inhibitory product due to the formation of a nonproductive SNARE complex(es) at release sites , whereas deletion of a further 17 amino acids permits replenishment and a speedy recovery . Inhibition of neurotransmitter release by peptides that mimic the N-terminal domain of P60880 . Botulinum neurotoxin serotypes A and E ( DB00083 and BoNT/E ) block neurotransmitter release by cleaving the 206-amino-acid SNARE protein , P60880 . For each BoNT serotype , cleavage of P60880 results in the loss of intact protein , the production of an N-terminal truncated protein , and the generation of a small C-terminal peptide . Peptides that mimic the C-terminal fragments of P60880 following DB00083 or BoNT/E cleavage were shown to depress transmitter release in bovine chromaffin cells and in Aplysia buccal ganglion cells . Similarly , the N-terminal-truncated P60880 resulting from DB00083 or BoNT/E cleavage has been found to inhibit transmitter exocytosis in various systems . With one exception , however , the inhibitory action of truncated P60880 has not been demonstrated at a well-defined cholinergic synapse . The goal of the current study was to determine the level of inhibition of neurotransmitter release by N-terminal DB00083 - or BoNT/E-truncated P60880 in two different neuronal systems : cholinergically coupled Aplysia neurons and rat hippocampal cell cultures . Both truncated P60880 products inhibited depolarization-dependent glutamate release from hippocampal cultures and depressed synaptic transmission in Aplysia buccal ganglion cells . These results suggest that truncated P60880 can compete with endogenous P60880 for binding with other SNARE proteins involved in transmitter release , thus inhibiting neurotransmitter exocytosis . Recombinant holotoxoid vaccine against botulism . The botulinum neurotoxins ( BoNT ) are the most toxic proteins for humans and designated " Category A Select Agents. " The current vaccine against botulism is in limited supply , and there is a need to develop new vaccine strategies . A recombinant DB00083 toxoid was produced in Clostridium botulinum that contained a double amino acid substitution , R363A Y365F ( termed DB00083 (RYM) ) . DB00083 (RYM) was noncatalytic for P60880 and nontoxic for mice . Immunization with DB00083 (RYM) protected mice from challenge at levels that were similar to chemically inactivated DB00083 toxoid . DB00083 (RYM) elicited an immune response against the light-chain and heavy-chain components of the toxin . Neutralizing anti- DB00083 (RYM) sera blocked BoNT toxicity in primary cortical neurons and blocked ganglioside binding by the heavy chain . DB00083 (RYM) represents a viable vaccine candidate for a holotoxoid against botulism . Characterization of Förster resonance energy transfer in a botulinum neurotoxin protease assay . Our previous article described a fluorescence-based assay for monitoring the proteolytic activity of botulinum neurotoxin types A and E ( DB00083 and BoNT/E ) . As detailed in that article , the assay is based on depolarization due to Förster resonance energy transfer between blue fluorescent protein ( Q9ULX5 ) and green fluorescent protein ( GFP ) moieties linked via residues 134-206 of P60880 ( synaptosome-associated protein of 25kDa ) , the protein substrate for DB00083 and BoNT/E . Before cleavage of this recombinant substrate , the polarization observed for the GFP emission , excited near the absorption maximum of the Q9ULX5 , is very low due to depolarization following energy transfer from Q9ULX5 to GFP . After substrate cleavage and diffusion of the fluorescent proteins beyond the energy transfer distance , the polarization is high due to observation of the emission only from directly excited GFP . This change in fluorescence polarization allows an assay , termed DARET ( depolarization after resonance energy transfer ) , that is robust and sensitive . In this article , we characterize the spectroscopic parameters of the system before and after substrate cleavage , including excitation and emission spectra , polarizations , and lifetimes . Mastoparan-7 rescues botulinum toxin-A poisoned neurons in a mouse spinal cord cell culture model . Botulinum neurotoxin serotype A ( DB00083 ) is the most potent poison of biological origin known to mankind . The toxicity of DB00083 is due to the inhibition of neurotransmission at cholinergic synapses ; this is responsible for the symptom of flaccid paralysis at peripheral neuromuscular junctions . At a molecular level , the DB00083 effect is due to its inhibition of stimulated acetylcholine ( ACh ) release from presynaptic nerve terminals . Currently , there is no antidote available to rescue DB00083 -poisoned synapses . Here , we report an example of rescuing botulinum-poisoned cultured mouse spinal cord neurons by treatment with Mastoparan-7 ( Mas-7 ) , which is known to be a phospholipase A2 activator compound . Mas-7 , a naturally occurring bee venom peptide , was delivered to botulinum-poisoned neurons via a drug delivery vehicle ( DDV ) construct prepared using the recombinant non-toxic heavy chain ( HC ) fragment of DB00083 itself . In this method , the DB00083 HC component in the DDV served as a neuron specific drug targeting molecule . We found that Mas-7 delivered into DB00083 intoxicated spinal cord cells restored over 40 % their property of stimulated neurotransmitter release . Rescue from cell poisoning did not occur from inhibition of the endopeptidase activity of DB00083 light chain ( LC ) against its well-known substrate , P60880 that is mechanistically involved in the cholinergic neuroexocytosis process . Rather , Mas-7 induced a physiological host response apparently unrelated to P60880 , but linked to the phospholipase-mediated signal transduction pathway . Probing DB00083 protease exosites : implications for inhibitor design and light chain longevity . Botulinum neurotoxin serotype A ( DB00083 ) is one of the most lethal toxins known . Its extreme toxicity is due to its light chain ( LC ) , a zinc protease that cleaves P60880 , a synaptosome-associated protein , leading to the inhibition of neuronal activity . Studies on DB00083 LC have revealed that two regions , termed exosites , can play an important role in BoNT catalytic activity . A clear understanding of how these exosites influence neurotoxin catalytic activity would provide a critical framework for deciphering the mechanism of P60880 cleavage and the design of inhibitors . Herein , based on the crystallographic structure of DB00083 LC complexed with its substrate , we designed an α-exosite binding probe . Experiments with this unique probe demonstrated that α-exosite binding enhanced both catalytic activity and stability of the LC . These data help delineate why α-exosite binding is needed for P60880 cleavage and also provide new insights into the extended lifetime observed for DB00083 LC in vivo . 8-Hydroxyquinoline and hydroxamic acid inhibitors of botulinum neurotoxin DB00083 . We describe here the state of the art of certain aspects concerning potential small molecule therapy directed toward botulism , by inhibition of the zinc-protease containing light chain ( LC ) of botulinum neurotoxin DB00083 from the anaerobic bacillus Clostridium botulinum . Botulinum neurotoxins ( BoNTs ) are comprised of eight serologically-distinct proteins ( A - H ) , several of which are further divided , such as DB00083 which has five subtypes . The BoNTs are the most toxic substances known to mankind , causing a form of flaccid paralysis that can be rapid and is often lethal . DB00083 is comprised of a ~100 kDa heavy chain ( HC ) attached via a single disulfide DB00151 - DB00151 bond to a ~50 kDa LC . The HC mediates transport to and uptake by presynaptic glutamatergic neurons , where the LC cleaves the protein P60880 and thus prevents vesicular trafficking and release of acetylcholine . The Zn-endoprotease activity of the LC of DB00083 is a target for the development of small molecule inhibitors of DB00083 -mediated toxicity . A variety of DB00083 LC inhibitors have been described to date and we focus here primarily on the Zn-binding 8-hydroxyquinoline structural type as well as some of the previously-described hydroxamic acids . DB00435 suppresses inducible nitric oxide synthase expression by inhibiting post-translational modification of IkappaB . The expression of inducible nitric oxide synthase ( P35228 ) is a critical factor in both normal physiological functions and the pathogenesis of disease . This study was undertaken to determine the molecular mechanism by which nitric oxide ( NO ) exerts negative feedback regulation on P35228 gene expression . Isolated rat hepatocytes stimulated with cytokines exhibited a marked increase in NO production as well as P35228 mRNA and protein levels , which were significantly reduced by pretreatment of the NO donors S-nitroso-N-acetyl-D,L-penicillamine ( P60880 ) and V-PYRRO/NO . This effect of P60880 was inhibited when NO was scavenged using red blood cells . Pretreatment with oxidized P60880 , 8-Br-cGMP , NO2- , or NO3- did not suppress the cytokine-induced NO production . Moreover , LPS/ P01579 -stimulated RAW264.7 cells , which produce endogenous NO , expressed lower levels of P35228 , IL-1beta , P05231 and P01375 mRNAs , without changes in their mRNA half-lives , than those in the presence of the P35228 inhibitor NG-monomethyl-L-arginine . The P35228 gene transcription rate exhibited an 18-fold increase after cytokine stimulation , which was significantly inhibited by P60880 pretreatment . P60880 also blocked cytokine- induced increase in NF-kappaB activation , P35228 promoter activity , nuclear translocation of cytosolic NF-kappaB p65 subunit , and P25963 degradation , which correlated with its inhibitory effect on phosphorylation and ubiquitination of IkappaB . These data indicate that NO down-regulates P35228 gene expression and NO production by inhibiting the post-translational processes of P25963 thereby preventing NF-kappaB activation . These results identify a novel negative feedback mechanism whereby NO down-regulates P35228 gene expression . Adeno-associated virus transfer of a gene encoding P60880 resistant to botulinum toxin A attenuates neuromuscular paralysis associated with botulism . Advances in viral gene therapy have opened new possibilities for treating a range of motor neuron diseases , but these have not yet been translated into clinically applicable therapies because of difficulties in delivery to susceptible/damaged neurons , ambiguities in the identity of gene(s) implicated , and a paucity of means to quantify any physiological improvement . Most of these hurdles can be overcome by using the neuromuscular paralysis induced by botulinum neurotoxin type A ( DB00083 ) as a prototype disease . Furthermore , because human botulism , occasionally fatal , causes prolonged muscle disablement as a result of the intraneuronal persistence of the toxin 's P60880 ( S25 ) -cleaving protease , development of a genetic approach could lead to a potential treatment for this debilitating disease . Adeno-associated viral delivery of a cleavage-resistant S25 gene ( S25-R198T ) to chromaffin cells in vitro yielded exocytotically active S25-R198T that diminished subsequent blockade by DB00083 of evoked catecholamine release . Evaluation in vivo , by administering this virus into rat spinal cord before injecting DB00083 , showed a decreased inhibition of acetylcholine release as reflected in elevated retention of neuromuscular transmission . A similar , although smaller , protection of synaptic transmission from the toxin was seen after peripherally injecting the therapeutic virus . Such therapy also curtailed nerve sprouting normally induced by DB00083 . This first demonstration of the utility of a DNA-based therapy for botulism paves the way for further advances in its treatment and for application to genetic disorders of motor neurons . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Is nitric oxide involved in P41595 receptor-mediated contraction in the rat stomach fundus ? Although contraction of the rat stomach fundus by 5-HT is known to be mediated by the P41595 receptor , the second messenger pathways involved in this response remain unclear . Since nitric oxide ( NO ) has been associated with contraction of certain gastrointestinal smooth muscle , the purpose of this study was to determine if NO is involved in 5-HT-induced contraction in the rat stomach fundus . The arginine analogs L-NAME and L-NMMA , at a concentration ( 100 microM ) established to inhibit NO synthase in the rat stomach fundus by inhibiting depolarization-induced relaxation in this tissue , had no effect on 5-HT contraction . Furthermore , the NO donors sodium nitroprusside and P60880 did not contract rat stomach fundus under basal tone , whereas 5-HT was a potent contractile agonist . These data do not support a role for NO in P41595 receptor-mediated contraction in the rat stomach fundus . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Peptide inhibitors of botulinum neurotoxin by mRNA display . Botulinum neurotoxins ( BoNTs ) are extremely toxic . The metalloproteases associated with the toxins cleave proteins essential for neurotransmitter secretion . Inhibitors of the metalloprotease are currently sought to control the toxicity of BoNTs . Toward that goal , we produced a synthetic cDNA for the expression and purification of the metalloprotease of DB00083 in Escherichia coli as a biotin-ubiquitin fusion protein , and constructed a combinatorial peptide library to screen for DB00083 light chain inhibitors using mRNA display . A protease assay was developed using immobilized intact P60880 as the substrate . The new peptide inhibitors showed a 10-fold increase in affinity to DB00083 light chain than the parent peptide . Interestingly , the sequences of the new peptide inhibitors showed abundant hydrophobic residues but few hydrophilic residues . The results suggest that mRNA display may provide a general approach in developing peptide inhibitors of BoNTs . Molecular targets of botulinum toxin at the mammalian neuromuscular junction . The molecular targets of botulinum neurotoxins ( BoNTs ) are SNARE ( soluble N-ethylmaleimide-sensitive factor-attachment protein-receptor ) proteins necessary for neurotransmitter release . BoNT are powerful therapeutic agents in the treatment of numerous neurological disorders . The goals of this study were to ( 1 ) assess toxin diffusion by measuring substrate cleavage in adjacent and distant muscles , and ( 2 ) characterize the clinical course using SNARE protein chemistry . A small volume of DB00083 was injected unilaterally into the mouse gastrocnemius muscle . Motor impairment was limited to the toxin-treated limb . No systemic illness or deaths occurred . At five time points , a subset of mice were killed , and muscles from both hindlimbs , and the diaphragm , were collected . Protein samples were examined for changes in P60880 ( synaptosomal-associated protein of Mr = 25 kDa ) using immunochemistry . P60880 cleavage product was noted in the toxin-treated limb as early as 1 day postinjection and continued through day 28 . Onset and peak levels of substrate cleavage corresponded to the onset and peak clinical response . Cleavage was observed in adjacent and distant muscles , demonstrating that substrate cleavage is a sensitive indicator of toxin diffusion . Significant increases in full-length P60880 and vesicle-associated membrane protein II were evident early in the impaired limb and continued through day 28 . The increased SNARE protein most likely originates from nerve terminal sprouts . A high-density single-nucleotide polymorphism screen of 23 candidate genes in attention deficit hyperactivity disorder : suggesting multiple susceptibility genes among Chinese Han population . Attention deficit hyperactivity disorder ( ADHD ) is a common childhood-onset behavioral disorder with a definite genetic component . The search for genes predisposing to ADHD has focused on genes involved in the regulation of monoamine systems . In this study , we emphasized genes that underlie various aspects of dopamine , norepinephrine and serotonin neurotransmissions and performed a comprehensive association analysis by screening with 245 single-nucleotide polymorphisms ( SNPs ) of 23 candidate genes in a sample of Chinese Han descent . A total of 182 DSM-IV ADHD children and 184 healthy controls were genotyped and analyzed with an average density of one SNP every 6.1 kb . Both single-SNP and multi-marker haplotype analyses were implemented to exploit association signal for ADHD and its diagnostic subtypes . Empirical P-values were derived on the basis of 5000 permutations to evaluate gene-wide statistical significance . P21397 yielded highly suggestive evidence of association ( empirical P < 0.01 , OR=1.94 ) with ADHD . For inattentive ADHD , P21397 , DDC and P08247 showed suggestive evidence of association ( empirical P < 0.05 ) . P18825 achieved suggestive significance ( empirical P < 0.05 ) for ADHD combined type . Additionally , for six genes ( P60880 , NET1 , P09172 , P43681 , P35462 and P21579 ) we detected one or more SNPs with nominal P-values </= 0.05 . This study has identified several genes as promising susceptibility loci for ADHD . Replication efforts and further investigations remain necessary to provide definite proof of association . Long-distance retrograde effects of botulinum neurotoxin A . Botulinum neurotoxins ( designated DB00083 -BoNT/G ) are bacterial enzymes that block neurotransmitter release by cleaving essential components of the vesicle fusion machinery . DB00083 , which cleaves P60880 ( synaptosomal-associated protein of 25 kDa ) , is extensively exploited in clinical medicine to treat neuromuscular pathologies , facial wrinkles , and various types of pain . It is widely assumed that DB00083 remains at the synaptic terminal and its effects are confined to the injection site . Here we demonstrate that catalytically active DB00083 is retrogradely transported by central neurons and motoneurons and is then transcytosed to afferent synapses , in which it cleaves P60880 . P60880 cleavage by DB00083 was observed in the contralateral hemisphere after unilateral DB00083 delivery to the hippocampus . Appearance of cleaved P60880 resulted in blockade of hippocampal activity in the untreated hemisphere . Injections of DB00083 into the optic tectum led to the appearance of DB00083 -truncated P60880 in synaptic terminals within the retina . Cleaved P60880 also appeared in the facial nucleus after injection of the toxin into rat whisker muscles . Experiments excluded passive spread of the toxin and demonstrated axonal migration and neuronal transcytosis of DB00083 . These findings reveal a novel pathway of DB00083 trafficking in neurons and have important implications for the clinical uses of this neurotoxin . Botulinum neurotoxin type D enables cytosolic delivery of enzymatically active cargo proteins to neurones via unfolded translocation intermediates . Multi-domain bacterial protein toxins are being explored as potential carriers for targeted delivery of biomolecules . Previous approaches employing isolated receptor binding subunits disallow entry into the cytosol . Strategies in which catalytic domains are replaced with cargo molecules are presumably inefficient due to co-operation of domains during the endosomal translocation step . Here , we characterize a novel transport vehicle in which cargo proteins are attached to the amino terminus of the full-length botulinum neurotoxin type D ( BoNT/D ) . The intrinsic enzymatic activity of the neurotoxin allowed quantification of the efficacy of cargo delivery to the cytosol . P00374 and BoNT type A ( DB00083 ) light chain ( LC ) were efficiently conveyed into the cytosol , whereas attachment of firefly luciferase or green fluorescent protein drastically reduced the toxicity . Luciferase and DB00083 LC retained their catalytic activity as evidenced by luciferin conversion or P60880 hydrolysis in the cytosol of synaptosomes , respectively . Conformationally stabilized dihydrofolate reductase as cargo considerably decreased the toxicity indicative for the requirement of partial unfolding of cargo protein and catalytic domain as prerequisite for efficient translocation across the endosomal membrane . Thus , enzymatically inactive clostridial neurotoxins may serve as effective , safe carriers for delivering proteins in functionally active form to the cytosol of neurones . Widespread sequence variations in P23763 across vertebrates suggest a potential selective pressure from botulinum neurotoxins . Botulinum neurotoxins ( DB00083 -G ) , the most potent toxins known , act by cleaving three SNARE proteins required for synaptic vesicle exocytosis . Previous studies on BoNTs have generally utilized the major SNARE homologues expressed in brain ( P63027 , syntaxin 1 , and P60880 ) . However , BoNTs target peripheral motor neurons and cause death by paralyzing respiratory muscles such as the diaphragm . Here we report that P23763 , but not P63027 , is the SNARE homologue predominantly expressed in adult rodent diaphragm motor nerve terminals and in differentiated human motor neurons . In contrast to the highly conserved P63027 , BoNT-resistant variations in P23763 are widespread across vertebrates . In particular , we identified a polymorphism at position 48 of P23763 in rats , which renders P23763 either resistant ( I48 ) or sensitive ( M48 ) to BoNT/D . Taking advantage of this finding , we showed that rat diaphragms with I48 in P23763 are insensitive to BoNT/D compared to rat diaphragms with M48 in P23763 . This unique intra-species comparison establishes P23763 as a physiological toxin target in diaphragm motor nerve terminals , and demonstrates that the resistance of P23763 to BoNTs can underlie the insensitivity of a species to members of BoNTs . Consistently , human P23763 contains I48 , which may explain why humans are insensitive to BoNT/D . Finally , we report that residue 48 of P23763 varies frequently between M and I across seventeen closely related primate species , suggesting a potential selective pressure from members of BoNTs for resistance in vertebrates . A structural perspective of the sequence variability within botulinum neurotoxin subtypes A1-A4 . Botulinum neurotoxin ( BoNT ) is a category A toxin that has been classified within seven serotypes , designated A-G . Recently , it has been discovered that sequence variability occurs in BoNTs produced by serotype A ( DB00083 ) variant strains , designated as subtypes A1 and A2 , which have significantly different antibody-binding properties . We have therefore made efforts to understand at the molecular level the diversity and its effects on the biological actions of the toxin , including receptor binding , substrate recognition , and catalysis . We provide the results of these studies , including the analysis of two newly sequenced DB00083 variants , Loch Maree ( A3 ) and 657Ba ( A4 ) , and their comparison to A1 and A2 . Using sequence analysis , available functional data , molecular modeling , and comparison of models with the crystal structures of BoNT/A1 and the light chain of BoNT/A2 , we conclude that these sequence differences within subtypes will impact development of broad-spectrum antibody and small ligand therapeutics , and suggest dissimilarities in binding affinity and cleavage efficiency of the P60880 substrate . In particular , sequence variation in subtypes BoNT/A3 and BoNT/A4 will likely effect alpha-exosite and S1 ' subsite recognition , respectively . P01308 / P05019 signaling pathways enhances tumor cell invasion through bisecting GlcNAc N-glycans modulation . an interplay with P12830 . Changes in glycosylation are considered a hallmark of cancer , and one of the key targets of glycosylation modifications is P12830 . We and others have previously demonstrated that P12830 has a role in the regulation of bisecting GlcNAc N-glycans expression , remaining to be determined the P12830 -dependent signaling pathway involved in this N-glycans expression regulation . In this study , we analysed the impact of P12830 expression in the activation profile of receptor tyrosine kinases such as insulin receptor ( IR ) and P08069 ( IGF-IR ) . We demonstrated that exogenous P12830 expression inhibits IR , IGF-IR and P29323 1/2 phosphorylation . Stimulation with insulin and P05019 in MDA-MD-435 cancer cells overexpressing P12830 induces a decrease of bisecting GlcNAc N-glycans that was accompanied with alterations on P12830 cellular localization . Concomitantly , IR/IGF-IR signaling activation induced a mesenchymal-like phenotype of cancer cells together with an increased tumor cell invasion capability . Altogether , these results demonstrate an interplay between P12830 and IR/IGF-IR signaling as major networking players in the regulation of bisecting N-glycans expression , with important effects in the modulation of epithelial characteristics and tumor cell invasion . Here we provide new insights into the role that P01308 / P05019 signaling play during cancer progression through glycosylation modifications . Detection of thymidylate synthase modulators by a novel screening assay . P04818 ( TS ) , a key cancer chemotherapeutic target , catalyzes the conversion of deoxyuridylate to thymidylate . TS can serve as a repressor of its own synthesis by binding to its own mRNA through TS-specific binding elements ( TBEs ) . In this report , we describe the use of a luciferase reporter plasmid containing two TBEs that can be used as a tool for the identification and initial profiling of compounds that modulate TS activity , levels , or ability to bind mRNA . To validate this model , we evaluated several groups of drugs . Thus , cells were exposed to the pyrimidine analogs 5-fluorouracil ( DB00544 ) , 5-fluorouridine ( DB01629 ) , 5-fluoro-2'-deoxyuridine ( FUdR ) , trifluorothymidine ( DB00432 ) ; to the nonpyrimidine TS-inhibitors AG-331 , nolatrexed ( AG337 ) , and raltitrexed ( DB00293 ) ; or to drugs with other primary sites of action ( methotrexate , actinomycin D , 5-azacytidine , 8-thioguanosine ) . Except for 5-azacytidine and 8-thioguanosine , all compounds examined induced luciferase activity compared with untreated cells . Effects of luciferase activity inducing drugs through TS-affected translation were confirmed by examinations of TS protein and mRNA levels . Treatment of H630- P13671 cells with DB00544 , DB01629 , FUdR , DB00432 , AG331 , AG337 , DB00293 , and methotrexate up-regulated TS levels as determined by Western blot analysis , although TS mRNA levels remained unchanged as determined by reverse transcription-polymerase chain reaction . Our studies demonstrate a novel application of a TBE-dependent reporter plasmid that could be used for the high-throughput identification of potential chemotherapeutic agents that modulate TS RNA-binding activity , either directly or indirectly . Chapter 3 : Molecular basis for the therapeutic effectiveness of botulinum neurotoxin type A . The utility of botulinum neurotoxin type A ( DB00083 ) for treating overactive muscles and endocrine glands is attributable to a unique conflation of properties honed to exploit and inactivate synaptic transmission . Specific , high-affinity coincident binding to gangliosides plus an intraluminal loop of synaptic vesicle protein 2 ( SV2 ) by the heavy chain ( HC ) of DB00083 confers selectivity for presynaptic nerve terminals and subsequent uptake by endocytosis . Upon vesicle acidification , the HC forms a channel for transmembrane transfer of the light chain to the cytosol , as observed by single channel recordings . The light chain is a Zn(2+) -dependent endoprotease that cleaves and inactivates P60880 , thereby blocking exocytotic release of transmitters , a discovery that revealed the pivotal role of the latter in synaptic vesicle fusion . A di-leucine motif in DB00083 light chain stabilizes this protease , contributing to its longevity inside nerves . The ubiquity of SV2 and P60880 has prompted re-evaluation of the nerve types susceptible to DB00083 . In urology , there is emerging evidence that DB00083 blocks neuropeptide release from afferent nerves , exocytosis of acetylcholine and purines from efferent nerves , and possibly DB00171 release from the urothelium . Suppression by DB00083 of the surface expression of nociceptor channels on bladder afferents might also contribute to its improvement of urological sensory symptoms . DB00435 induces apoptosis in GM- P04141 -treated eosinophils via caspase-6-dependent lamin and DNA fragmentation . Asthma is characterized by accumulation of eosinophils in the lungs and delayed apoptosis may be one mechanism leading to eosinophilia . DB00435 ( NO ) , present in inflamed lungs , has been shown to possess both anti- and proeosinophilic properties . We previously showed that NO induces apoptosis in the presence of survival prolonging cytokine P05113 in human eosinophils . In the present study , we examined the intracellular mechanisms of NO-induced apoptosis in granulocyte macrophage-colony stimulating factor ( GM- P04141 ) -treated eosinophils concentrating on the role of caspases and calpains . Eosinophils were isolated from human blood and apoptosis was determined by relative DNA fragmentation assay , morphological analysis and/or Annexin-V FITC assay . We showed that NO-donor S-nitroso-N-acetyl-d,l-penicillamine ( P60880 ) induced apoptosis in GM- P04141 -treated eosinophils . P60880 -induced DNA fragmentation was totally prevented by an inhibitor of caspase-6 ( Z-VEID-FMK ) . Decreased levels of caspase-6 proenzyme and increased amounts of cleaved lamin A/C in P60880 -treated cells indicated activation of caspase-6 . Furthermore , P60880 -induced lamin A/C and B fragmentation was totally abolished by an inhibitor of caspase-6 . According to our results , caspase-6 mediates lamin and DNA fragmentation also in spontaneously dying eosinophils . Inhibitor of calpains prevented most of DNA fragmentation related to spontaneous apoptosis but had no effect in eosinophils undergoing NO-induced apoptosis . In the present study we showed that caspase-6 is essential for the executive phase involving lamin and DNA fragmentation in both NO-induced and spontaneous eosinophil apoptosis . However , differences in the involvement of calpains suggest that the intracellular signalling in NO-induced apoptosis has specific features at the level of proteases . This study demonstrates new mechanisms for NO-induced and spontaneous apoptosis in human eosinophils . Tandem fluorescent proteins as enhanced FRET-based substrates for botulinum neurotoxin activity . The light chain of botulinum neurotoxin A ( DB00083 -LC ) is a zinc-metalloprotease that requires two extended exosites for optimal substrate binding and recognition of its intracellular target P60880 . P27918 and YFP connected through P60880 peptide ( 141-206 ) containing both exosites ( CsY ) has been used in a FRET-based assay for DB00083 . To further improve the FRET efficiency in this DB00083 substrate for in vitro high-throughput assays , we explored the feasibility of enhancing the capture of P27918 emission by doubling the number of YFP acceptors . In comparison to CsY , the tandem fluorescence substrates CsYY and YsCsY enhanced the ratiometric fluorescence signal between YFP and P27918 . YsCsY , containing two substrate sites , offered the greatest fluorometric change upon toxin-catalyzed cleavage . In addition to known approaches for enhancing fluorescence yield through various mutations , this alternative tandem substrate approach can boost the FRET signal and is particularly useful for substrates requiring extensive exosite recognition for specificity . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Therapeutic effectiveness of botulinum neurotoxin A : potent blockade of autonomic transmission by targeted cleavage of only the pertinent P60880 . In search of a basis for the impressive potency of an endoprotease that cleaves P60880 , botulinum neurotoxin type A ( DB00083 ) , in treating numerous diseases due to hyper-active autonomic nerves , truncation of its target and inhibition of neurotransmission were studied in rat sympathetic neurons . DB05232 -sensitive spontaneous cholinergic neurotransmission was blocked > 80 % by 1 pM DB00083 despite cleaving < 20 % of the P60880 . A maximum cleavage of ∼60 % P60880 could be achieved with > 1 nM DB00083 , despite an absence of non-cleavable P60880 in the detergent-solubilised neurons . In contrast , BoNT/E ( 100 nM ) truncated nearly all the P60880 in the intact cells , but was unable to block neurotransmission at low concentrations like DB00083 . Chimeras created by inserting the acceptor-binding HC domain of DB00083 into BoNT/E still cleaved all the P60880 , indicating ubiquitous expression of DB00083 acceptors . Accordingly , SV2 and P60880 were found to be co-expressed and broadly co-localised in neurons , but absent from non-neuronal cells . On the other hand , partial cleavage by the DB00083 protease persisted upon replacing its HC with counterparts from BoNT/E or BoNT/B . Moreover , limited cleavage of P60880 was conferred onto the protease from BoNT/E when fused to the N-terminus of DB00083 . Thus , the DB00083 protease is uniquely well-adapted for selectively inactivating the P60880 directly involved in neurotransmission ; this together with the toxin 's acceptor and its target being localised on the peri-somatic boutons likely contribute to its exceptional therapeutic utility in the clinic . Prevalence of borreliosis , anaplasmosis , ehrlichiosis and Dirofilaria immitis in dogs and vectors in Voronezh Reserve ( Russia ) . Most of the dogs studied for the prevalence of CVBD have previously received acaricidal and insecticidal treatments . In the present work , a very specific population of dogs ( Group 1 ) that had never been treated against ticks and mosquitoes was studied . Moreover , the territory occupied by this population has also never been treated , because it is a protected area -- Voronezh Natural Reserve . Canine patients from veterinary clinics ( Group 2 ) that had been treated against VBD vectors were studied for comparison . Eighty-two dogs ( Group 1 ) were enrolled in June , 2008 . Blood samples were tested using the IDEXX P60880 (®) 4Dx(®) test . A specific heartworm antigen was detected in 12.2 % samples . The seroprevalence for Anaplasma phagocytophilum was found to be 34.1 % . The antibodies to Borrelia P13671 peptide and to Ehrlichia canis were detected in 2.4 % of the samples . Almost all dogs with infections had no clinical signs . Only 3 mixed-infected dogs showed non-specific clinical signs . During the tick season , 358 Ixodes ricinus were collected ; the prevalence of Borrelia burgdorferi s.l. and Anaplasma phagocytophilum was 21.9 % and 0.6 % , respectively . Four hundred and forty dogs ( Group 2 ) were studied for comparison . Antibodies to B. burgdorferi s.l. were detected only in one dog , seroprevalence for A. phagocytophilum represented 1.1 % , no E. canis seropositive dogs were identified , and 8.2 % dogs were found infected with Dirofilaria immitis . Fifty-six percent of dogs with dirofilariosis had clinical signs . All dogs with anaplasmosis showed specific clinical signs -- fever , anemia , splenitis . Three dogs died within a few days . Primary cultures of embryonic chicken neurons for sensitive cell-based assay of botulinum neurotoxin : implications for therapeutic discovery . Botulinum toxin is an exceedingly potent inhibitor of neurotransmission across the neuromuscular junction , causing flaccid paralysis and death . The potential for misuse of this deadly poison as a bioweapon has added a greater urgency to the search for effective therapeutics . The development of sensitive and efficient cell-based assays for the evaluation of toxin antagonists is crucial to the rapid and successful identification of therapeutic compounds . The authors evaluated the sensitivity of primary cultures from 4 distinct regions of the embryonic chick nervous system to botulinum neurotoxin A ( DB00083 ) cleavage of synaptosomal-associated protein of 25 kD ( P60880 ) . Although differences in sensitivity were apparent , P60880 cleavage was detectable in neuronal cells from each of the 4 regions within 3 h at DB00083 concentrations of 1 nM or lower . Co-incubation of chick neurons with DB00083 and toxin-neutralizing antibodies inhibited P60880 cleavage , demonstrating the utility of these cultures for the assay of DB00083 antagonists . DB00435 -cGMP-protein kinase G signaling pathway induces anoxic preconditioning through activation of DB00171 -sensitive K+ channels in rat hearts . DB00435 ( NO ) plays an important role in anoxic preconditioning to protect the heart against ischemia-reperfusion injuries . The present work was performed to study better the NO-cGMP-protein kinase G ( PKG ) signaling pathway in the activation of both sarcolemmal and mitochondrial DB00171 -sensitive K+ ( KATP ) channels during anoxic preconditioning ( P25054 ) and final influence on reducing anoxia-reperfusion ( A/R ) -induced cardiac damage in rat hearts . The upstream regulating elements controlling NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection were investigated . The involvement of both inducible and endothelial NO synthases ( P35228 and P29474 ) in the progression of this signaling pathway was followed . Final cellular outcomes of ischemia-induced injury after different preconditioning in the form of lactate dehydrogenase release , DNA strand breaks , and malondialdehyde formation as indexes of cell injury and lipid peroxidation , respectively , were investigated . The lactate dehydrogenase and malondialdehyde values decreased in the groups that underwent preconditioning periods with specific mitochondrial KATP channels opener diazoxide ( 100 microM ) , nonspecific mitochondrial KATP channels opener pinacidil ( 50 microM ) , S-nitroso-N-acetylpenicillamine ( P60880 , 300 microM ) , or beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclicmonophosphorothioate , Sp-isomer ( 10 microM ) before the A/R period . Preconditioning with P60880 significantly reduced the DNA damage . The effect was blocked by glibenclamide ( 50 microM ) , 5-hydroxydecanoate ( 100 microM ) , NG-nitro-L-arginine methyl ester ( 200 microM ) , and beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclic monophosphorothioate , Rp-isomer ( 1 microM ) . The results suggest P35228 , rather than P29474 , as the major contributing NO synthase during P25054 treatment . Moreover , the PKG shows priority over NO as the upstream regulator of NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection during P25054 treatment . Antagonism of botulinum toxin type A-induced cleavage of P60880 in rat cerebral synaptosome by toosendanin . Toosendanin ( Q15631 ) , a triterpenoid derivative extracted from Chinese traditional medicine , has been demonstrated to be an effective cure for experimental botulism . This study is designed to explore its antibotulismic mechanism by Western blotting . The results showed that Q15631 incubation did not change the electrophoresis pattern and the amounts of synaptosomal-associated protein of 25 kDa ( P60880 ) , syntaxin and synaptobrevin/vesicle-associated membrane protein in rat cerebral synaptosomes , but made the synaptosomes completely resistant to botulinum neurotoxin A ( DB00083 ) -mediated cleavage of P60880 . After binding of DB00083 to synaptosomes , Q15631 still partially antagonized the toxin-mediated cleavage of P60880 . However , Q15631 -incubated synaptosomal membrane fraction did not resist the cleavage of P60880 by the light chain of DB00083 . It is suggested that the antibotulismic effect of Q15631 results from blocking the toxin 's approach to its enzymatic substrate . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . The hyh mutation uncovers roles for alpha Snap in apical protein localization and control of neural cell fate . The hyh ( hydrocephalus with hop gait ) mouse shows a markedly small cerebral cortex at birth and dies postnatally from progressive enlargement of the ventricular system . Here we show that the small hyh cortex reflects altered cell fate . Neural progenitor cells withdraw prematurely from the cell cycle , producing more early-born , deep-layer cerebral cortical neurons but depleting the cortical progenitor pool , such that late-born , upper-layer cortical neurons are underproduced , creating a small cortex. hyh mice carry a hypomorphic missense mutation in the gene Napa encoding soluble N-ethylmaleimide-sensitive factor ( P46459 ) attachment protein alpha ( alpha Snap ) , involved in P60880 receptor ( SNARE ) -mediated vesicle fusion in many cellular contexts . A targeted null Napa mutation is embryonically lethal . Altered neural cell fate is accompanied by abnormal localization of many apical proteins implicated in regulation of neural cell fate , including P12830 , beta-catenin , atypical protein kinase C ( aPKC ) and Q8NI35 ( inactivation-no-afterpotential D-like , also known as protein associated with Lin7 , or Pals1 ) . Apical localization of the SNARE Vamp7 is also disrupted . Thus , alpha Snap is essential for apical protein localization and cell fate determination in neuroepithelial cells . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . DB00368 inhibits exocytosis via the G protein βγ subunit and refilling of the readily releasable granule pool via the α(i1/2) subunit . The molecular mechanisms responsible for the ' distal ' effect by which noradrenaline ( NA ) blocks exocytosis in the β-cell were examined by whole-cell and cell-attached patch clamp capacitance measurements in P01308 832/13 β-cells . NA inhibited Ca(2+)-evoked exocytosis by reducing the number of exocytotic events , without modifying vesicle size . Fusion pore properties also were unaffected . NA-induced inhibition of exocytosis was abolished by a high level of Ca(2+) influx , by intracellular application of antibodies against the G protein subunit Gβ and was mimicked by the myristoylated βγ-binding/activating peptide mSIRK . NA-induced inhibition was also abolished by treatment with DB00083 , which cleaves the C-terminal nine amino acids of P60880 , and also by a P60880 C-terminal-blocking peptide containing the DB00083 cleavage site . These data indicate that inhibition of exocytosis by NA is downstream of increased [Ca(2+)](i) and is mediated by an interaction between Gβγ and the C-terminus of P60880 , as is the case for inhibition of neurotransmitter release . Remarkably , in the course of this work , a novel effect of NA was discovered . NA induced a marked retardation of the rate of refilling of the readily releasable pool ( O75783 ) of secretory granules . This retardation was specifically abolished by a Gα(i1/2) blocking peptide demonstrating that the effect is mediated via activation of Gα(i1) and/or Gα(i2) .

MH_train_3

MH_train_3

MH_train_3

interacts_with DB00083?

multiple_choice

A protein chip membrane-capture assay for botulinum neurotoxin activity . Botulinum neurotoxins A and B ( DB00083 and B ) are neuromuscular blocking agents which inhibit neurotransmission by cleaving the intra-cellular presynaptic SNARE proteins P60880 and P63027 , localized respectively in plasma membrane and synaptic vesicles . These neurotoxins are both dangerous pathogens and powerful therapeutic agents with numerous clinical and cosmetic applications . Consequently there is a need for in vitro assays of their biological activity to screen for potential inhibitors and to replace the widely used in vivo mouse assay . Surface plasmon resonance ( SPR ) was used to measure membrane vesicle capture by antibodies against P60880 and P63027 . Substrate cleavage by BoNTs modified capture providing a method to assay toxin activity . Firstly using synaptic vesicles as a substrate , a comparison of the EC(50)s for BoNT/B obtained by SPR , ELISA or flow cytometry indicated similar sensitivity although SPR assays were more rapid . Sonication of brain or neuronal cultures generated plasma membrane fragments with accessible intra-cellular epitopes adapted to measurement of DB00083 activity . SPR responses were proportional to antigen concentration permitting detection of as little as 4 pM P60880 in crude lysates . DB00083 activity was assayed using monoclonal antibodies that specifically recognize a P60880 epitope generated by the proteolytic action of the toxin . Incubation of intact primary cultured neurons with DB00083 yielded an EC(50) of 0.5 pM . The SPR biosensor method was sensitive enough to monitor DB00083 and B activity in cells cultured in a 96-well format providing an alternative to experimental animals for toxicological assays . Peptide inhibitors of botulinum neurotoxin by mRNA display . Botulinum neurotoxins ( BoNTs ) are extremely toxic . The metalloproteases associated with the toxins cleave proteins essential for neurotransmitter secretion . Inhibitors of the metalloprotease are currently sought to control the toxicity of BoNTs . Toward that goal , we produced a synthetic cDNA for the expression and purification of the metalloprotease of DB00083 in Escherichia coli as a biotin-ubiquitin fusion protein , and constructed a combinatorial peptide library to screen for DB00083 light chain inhibitors using mRNA display . A protease assay was developed using immobilized intact P60880 as the substrate . The new peptide inhibitors showed a 10-fold increase in affinity to DB00083 light chain than the parent peptide . Interestingly , the sequences of the new peptide inhibitors showed abundant hydrophobic residues but few hydrophilic residues . The results suggest that mRNA display may provide a general approach in developing peptide inhibitors of BoNTs . Enhancement of the endopeptidase activity of botulinum neurotoxin by its associated proteins and dithiothreitol . Botulinum neurotoxins type A ( DB00083 ) , the most toxic substance known to man , is produced by Clostridium botulinum type A as a complex with a group of neurotoxin-associated proteins ( NAPs ) , possibly through a polycistronic expression of a clustered group of genes . The botulinum neurotoxin complex is the only known example of a protein complex where a group of proteins ( NAPs ) protect another protein ( BoNT ) against acidity and proteases of the GI tract . We now report that NAPs also potentiate the DB01593 endopeptidase activity of DB00083 in both in vitro and in vivo assays against its known intracellular target protein , 25 kDa synaptosomal associated protein ( P60880 ) . While DB00083 exhibited no protease activity prior to reduction with dithiothreitol ( DTT ) , the DB00083 complex exhibited a high protease activity even in its nonreduced form . Our results suggest that the bacterial production of NAPs along with BoNT is designed for the NAPs to play an accessory role in the neurotoxin function , in contrast to their previously known limited role in protecting the neurotoxin in the GI tract and in the external environment . Structural features of DB00083 change considerably upon disulfide reduction , as revealed by near-UV circular dichroism spectroscopy . DB00083 in the reduced form adopts a more flexible structure than in the unreduced form , as also indicated by large differences in DeltaH values ( 155 vs 248 kJ mol-1 ) of temperature-induced unfolding of DB00083 . Inhibition of neurotransmitter release by clostridial neurotoxins correlates with specific proteolysis of synaptosomal proteins . Rat brain synaptosomes were used to study the effect of several clostridial neurotoxins on the neurotransmitter release . In this system the blockade of transmitter release correlated with the proteolytic activity of the toxins . Blockade of glutamate release was linked to selective proteolysis of one of the following synaptic proteins : synaptobrevin ( BoNT/D , BoNT/F ) ; P60880 ( DB00083 , BoNT/E ) , or HPC-1/syntaxin ( BoNT/C1 ) . All the toxins used had an inhibitory effect on synaptosomes with the exception of BoNT/F . BoNT/F cleaved synaptobrevin in permeabilized synaptosomes but failed to produce the same effect on intact synaptosomes . Calorie restriction promotes mammalian cell survival by inducing the Q96EB6 deacetylase . A major cause of aging is thought to result from the cumulative effects of cell loss over time . In yeast , caloric restriction ( CR ) delays aging by activating the Sir2 deacetylase . Here we show that expression of mammalian Sir2 ( Q96EB6 ) is induced in CR rats as well as in human cells that are treated with serum from these animals . P01308 and insulin-like growth factor 1 ( DB01277 ) attenuated this response . Q96EB6 deacetylates the DNA repair factor P12956 , causing it to sequester the proapoptotic factor Bax away from mitochondria , thereby inhibiting stress-induced apoptotic cell death . Thus , CR could extend life-span by inducing Q96EB6 expression and promoting the long-term survival of irreplaceable cells . The destructive effect of botulinum neurotoxins on the SNARE protein : P60880 and synaptic membrane fusion . Synaptic exocytosis requires the assembly of syntaxin 1A and P60880 on the plasma membrane and synaptobrevin 2 ( P63027 ) on the vesicular membrane to bridge the two opposite membranes . It is believed that the three SNARE proteins assemble in steps along the dynamic assembly pathway . The C-terminus of P60880 is known to be the target of botulinum neurotoxins ( DB00083 and BoNT/E ) that block neurotransmitters release in vivo . In this study , we employed electron paramagnetic resonance ( EPR ) spectroscopy to investigate the conformation of the P60880 C-terminus in binary and ternary SNARE complexes . The fluorescence lipid mixing assay shows that the C-terminal of P60880 is essential for membrane fusion , and that the truncated P60880 mutants cleaved by DB00083 and BoNT/E display different inhibition effects on membrane fusion : P60880 -25E ( Δ26 ) abolishes the fusion activity of the SNARE complex , while P60880 -25A ( Δ9 ) loses most of its function , although it can still form a SDS-resistant SNARE complex as the wild-type P60880 . CW-EPR spectra validate the unstable structures of the SNARE complex formed by P60880 mutants . We propose that the truncated P60880 mutants will disrupt the assembly of the SNARE core complex , and then inhibit the synaptic membrane fusion accordingly . Innocuous full-length botulinum neurotoxin targets and promotes the expression of lentiviral vectors in central and autonomic neurons . Fragments of botulinum neurotoxin ( BoNT ) have been explored as potential targeting moieties and carriers of biomolecules into neurons , although with lower binding and translocation efficiency compared with intact proteins . This study exploits a detoxified recombinant form of full-length BoNT/B ( BoTIM/B ) fused with core streptavidin ( CS-BoTIM/B ) for lentiviral targeting to central and autonomic neurons . CS-BoTIM/B underwent an activity-dependent entry into cultured spinal cord neurons . Coupling CS-BoTIM/B to biotinylated lentivirus-encoding green fluorescent protein ( GFP ) endowed considerable neuron selectivity to the vector as evident from the preferential expression of the reporter in neurons co-cultured with skeletal muscle cells . CS-BoTIM/B-guided lentiviral transduction with the expression of a SNARE protein , P60880 ( S25 ) , rendered non-susceptible to proteolysis by three BoNT serotypes , yielded a sizable decrease in cleaved S25 upon exposure of spinal cord neurons to these toxins . This was accompanied by synaptic transmission being spared from blockade by DB00083 or BoNT/E , reflecting adequate translation and functional competence of recombinant multi-toxin-resistant S25 . The augmented neurotropism conveyed on the lentivirus by CS-BoTIM/B was also demonstrated in vivo through enhanced expression of a reporter in intramural ganglionic neurons in the rat trachea , after injection of the targeted GFP-encoding lentivirus . Thus , a novel and realistic prospect for gene therapy of peripheral neuropathies is offered in this study through lentiviral targeting to neurons by CS-BoTIM/B . Behavioral and immunohistochemical evidence for central antinociceptive activity of botulinum toxin A . Botulinum toxin A ( DB00083 ) is approved for treatment of different cholinergic hyperactivity disorders , and , recently , migraine headache . Although suggested to act only locally , novel observations demonstrated bilateral reduction of pain after unilateral toxin injection , and proposed retrograde axonal transport , presumably in sensory neurons . However , up to now , axonal transport of DB00083 from periphery to CNS was identified only in motoneurons , but with unknown significance . We assessed the effects of low doses of DB00083 injected into the rat whisker pad ( 3.5 U/kg ) or into the sensory trigeminal ganglion ( 1 U/kg ) on formalin-induced facial pain . Axonal transport was prevented by colchicine injection into the trigeminal ganglion ( 5 mM , 2 μl ) . To find the possible site of action of axonally transported DB00083 , we employed immunohistochemical labeling of DB00083 -truncated synaptosomal-associated protein 25 ( P60880 ) in medullary dorsal horn of trigeminal nucleus caudalis after toxin injection into the whisker pad . Both peripheral and intraganglionic DB00083 reduce phase II of formalin-induced pain . Antinociceptive effect of DB00083 was prevented completely by colchicine . DB00083 -truncated P60880 in medullary dorsal horn ( spinal trigeminal nucleus ) was evident 3 days following the peripheral treatment , even with low dose applied ( 3.5 U/kg ) . Presented data provide the first evidence that axonal transport of DB00083 , obligatory for its antinociceptive effects , occurs via sensory neurons and is directed to sensory nociceptive nuclei in the CNS . Dynamics of motor nerve terminal remodeling unveiled using SNARE-cleaving botulinum toxins : the extent and duration are dictated by the sites of P60880 truncation . Nerve sprouts emerge from motor nerve terminals following blockade of exo-endocytosis for more than 3 days by botulinum neurotoxin ( BoNT ) , and form functional synapses , albeit temporary . Upon restoration of synaptic activity to the parent terminal 7 and 90 days after exposure to BoNT/F or A respectively , a concomitant retraction of the outgrowths was observed . BoNT/E caused short-term neuroparalysis , and dramatically accelerated the recovery of DB00083 -paralyzed muscle by further truncation of P60880 and its replenishment with functional full-length SNARE . The removal of 9 C-terminal residues from P60880 by DB00083 leads to persistence of the inhibitory product due to the formation of a nonproductive SNARE complex(es) at release sites , whereas deletion of a further 17 amino acids permits replenishment and a speedy recovery . Heterogeneity of genomic fusion of P11274 and P00519 in Philadelphia chromosome-positive acute lymphoblastic leukemia . Philadelphia chromosome-positive acute lymphoblastic leukemia occurs in two molecular forms , those with and those without rearrangement of the breakpoint cluster region on chromosome 22 . The molecular abnormality in the former group is similar to that found in chronic myelogenous leukemia . To characterize the abnormality in the breakpoint cluster region-unrearranged form , we have mapped a 9;22 translocation from the Philadelphia chromosome-positive acute lymphoblastic leukemia cell line P60880 -B13 by using pulsed-field gel electrophoresis and have cloned the DNA at the translocation junctions . We demonstrate a P11274 - P00519 fusion gene on the Philadelphia chromosome . The breakpoint on chromosome 9 is within P00519 between exons Ia and II , and the breakpoint on chromosome 22 is approximately equal to 50 kilobases upstream of a breakpoint cluster region in an intron of the P11274 gene . This upstream P11274 breakpoint leads to inclusion of fewer P11274 sequences in the fusion gene , compared with the P11274 - P00519 fusion gene of chronic myelogenous leukemia . Consequently , the associated mRNA and protein are smaller . The exons from P00519 are the same . Analysis of leukemic cells from four other patients with breakpoint cluster region-unrearranged Philadelphia chromosome-positive acute lymphoblastic leukemia revealed a rearrangement on chromosome 22 close to the breakpoint in P60880 -B13 in only one patient . These data indicate that breakpoints do not cluster tightly in this region but are scattered , possibly in a large intron . Given the large size of P11274 and the heterogeneity in breakpoint location , detection of P11274 rearrangement by standard Southern blot analysis is difficult . Pulsed-field gel electrophoresis should allow detection at the DNA level in every patient and thus will permit clinical correlation of the breakpoint location with prognosis . Botulinum neurotoxin C1 cleaves both syntaxin and P60880 in intact and permeabilized chromaffin cells : correlation with its blockade of catecholamine release . The seven types ( A -- G ) of botulinum neurotoxin ( BoNT ) are DB01593 -dependent endoproteases that potently block neurosecretion . Syntaxin is presently thought to be the sole substrate for BoNT/C1 , and synaptosomal-associated protein of Mr = 25 000 ( P60880 ) is selectively proteolyzed by types A and E . In this study , the effects of C1 on Ca2+ -regulated exocytosis of dense core granules from adreno-chromaffin cells were examined together with its underlying molecular action . Intact chromaffin cells were exposed to the toxin , and catecholamine release therefrom was then measured in conjunction with the monitoring of syntaxin cleavage by Western blotting . A good correlation was obtained between degradation of syntaxin 1A/B and reduction in Ca2+- or Ba2+-dependent secretion . However , blotting with antibodies against a C-terminal peptide of P60880 revealed the additional disappearance of immunoreactivity , with the same toxin concentration dependency as syntaxin breakdown . Notably , the cleaved P60880 product was similar in size to that produced by DB00083 ; however , contamination of BoNT/C1 by serotypes A or E was eliminated . Therefore , it is concluded that syntaxin 1A/B and P60880 are cleaved in intact cells poisoned with only C1 . Notably , C1 treatment of chromaffin cells abolished Ca2+ -evoked secretion following digitonin permeabilization , compared with partial inhibition by DB00083 , suggesting the importance of syntaxin for catecholamine release . Unexpectedly , C1 failed to proteolyze a soluble recombinant P60880 , even though it served as an efficient substrate for DB00083 . These interesting observations suggest that C1 can only efficiently cleave P60880 in intact cells , possibly due to the existence therein of a unique conformation and/or the participation of accessory factors . Involvement of P60880 in TRH-induced exocytosis in pituitary GH4C1 cells . The synaptic membrane protein synaptosomal-associated protein ( P60880 ) has recently been implicated as one of the key proteins involved in exocytotic membrane fusion in neurons . However , the role of P60880 in pituitary hormone release is not known . In this study , we determined that P60880 is involved in regulated exocytosis in the clonal pituitary cell line GH4C1 . P60880 messenger RNA and protein were detected in GH4C1 cells by RT-PCR and immunoblot analysis , respectively . Immunofluorescence analysis indicated that P60880 protein was localized in the plasma membrane . Next , to determine the function of P60880 in GH4C1 cells , specific inhibitors of P60880 , botulinum neurotoxin ( BoNT ) /A or /E , and antisense P60880 oligonucleotide were used . Neither DB00083 nor BoNT/E affected thyrotropin-releasing hormone ( TRH ) -induced cytosolic Ca2+ increase , but both inhibited TRH-induced exocytosis . Moreover , they dose-dependently inhibited TRH-induced prolactin release . The introduction of antisense oligonucleotide into the cells also inhibited TRH-induced prolactin release . These results suggest that P60880 is involved in regulated exocytosis in GH4C1 cells . Site-directed mutagenesis identifies active-site residues of the light chain of botulinum neurotoxin type A . Botulinum neurotoxins ( BoNTs ) are metalloproteases which block neuroexocytosis via specific cleavage and inactivation of SNARE proteins . Such proteolysis accounts for the extreme toxicity of these neurotoxins and of their prolonged effect . The recently determined structures of DB00083 and/B allows one to design active-site mutants to probe the role of specific residues in the proteolysis of SNARE proteins . Here we present the results of mutations of the second glutamyl residue involved in zinc coordination and of a tyrosine and a phenylalanine residues that occupy critical positions within the active site of DB00083 . The spectroscopic properties of the purified mutants are closely similar to those of the wild-type molecule indicating the acquisition of a correct tertiary structure . Mutation of the DB00142 -262* nearly abolishes P60880 hydrolysis as expected for a residue involved in zinc coordination . The DB00120 -266 and DB00135 -366 mutants have reduced proteolytic activity indicating a direct participation in the proteolytic reaction , and their possible role in catalysis is discussed . Cdk5/p35 regulates neurotransmitter release through phosphorylation and downregulation of P/Q-type voltage-dependent calcium channel activity . P12004 -dependent kinase 5 ( Cdk5 ) is a proline-directed serine/threonine kinase with close structural homology to the mitotic Cdks . The complex of Cdk5 and p35 , the neuron-specific regulatory subunit of Cdk5 , plays important roles in brain development , such as neuronal migration and neurite outgrowth . Moreover , Cdk5 is thought to be involved in the promotion of neurodegeneration in Alzheimer 's disease . Cdk5 is abundant in mature neurons ; however , its physiological functions in the adult brain are unknown . Here we show that Cdk5/p35 regulates neurotransmitter release in the presynaptic terminal . Both Cdk5 and p35 were abundant in the synaptosomes . Roscovitine , a specific inhibitor of Cdk5 in neurons , induced neurotransmitter release from the synaptosomes in response to membrane depolarization and enhanced the EPSP slopes in rat hippocampal slices . The electrophysiological study using each specific inhibitor of the voltage-dependent calcium channels ( VDCCs ) and calcium imaging revealed that roscovitine enhanced Ca2+ influx from the P/Q-type VDCC . Moreover , Cdk5/p25 phosphorylated the intracellular loop connecting domains II and III ( L(II-III) ) between amino acid residues 724 and 981 of isoforms cloned from rat brain of the alpha1A subunit of P/Q-type Ca2+ channels . The phosphorylation inhibited the interaction of L(II-III) with P60880 and synaptotagmin I , which were plasma membrane soluble N-ethylmaleimide-sensitive factor attachment protein ( P60880 ) receptor ( SNARE ) proteins and were required for efficient neurotransmitter release . These results strongly suggest that Cdk5/p35 inhibits neurotransmitter release through the phosphorylation of P/Q-type VDCC and downregulation of the channel activity . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Correlation of cleavage of P60880 with muscle function in a rat model of Botulinum neurotoxin type A induced paralysis . Injection of botulinum neurotoxin serotype A ( DB00083 ) into muscle results in cleavage of the synaptosomal associated protein of 25 kDa ( P60880 ) and relatively long-term paralysis . However , nerve-terminal sprouting , which appears to require intact P60880 , has been reported to occur much earlier . The difference between the long-term paralysis induced by injection of DB00083 and the short time needed for sprouting led us to investigate the relationship between DB00083 catalyzed cleavage of P60880 and muscle function . The effect of DB00083 on P60880 present in nerve endings innervating gastrocnemius muscles of rats was monitored over time . Cleaved P60880 was found in nerve terminals innervating the muscles within 24h of inoculation with DB00083 and was present more than 2 months later . Comparison of the ratios of cleaved to intact P60880 from the onset of DB00083 -induced paralysis until function was regained indicated that paralysis was probable when the ratio of cleaved to intact P60880 was greater than 0.35 . Chapter 3 : Molecular basis for the therapeutic effectiveness of botulinum neurotoxin type A . The utility of botulinum neurotoxin type A ( DB00083 ) for treating overactive muscles and endocrine glands is attributable to a unique conflation of properties honed to exploit and inactivate synaptic transmission . Specific , high-affinity coincident binding to gangliosides plus an intraluminal loop of synaptic vesicle protein 2 ( SV2 ) by the heavy chain ( HC ) of DB00083 confers selectivity for presynaptic nerve terminals and subsequent uptake by endocytosis . Upon vesicle acidification , the HC forms a channel for transmembrane transfer of the light chain to the cytosol , as observed by single channel recordings . The light chain is a Zn(2+) -dependent endoprotease that cleaves and inactivates P60880 , thereby blocking exocytotic release of transmitters , a discovery that revealed the pivotal role of the latter in synaptic vesicle fusion . A di-leucine motif in DB00083 light chain stabilizes this protease , contributing to its longevity inside nerves . The ubiquity of SV2 and P60880 has prompted re-evaluation of the nerve types susceptible to DB00083 . In urology , there is emerging evidence that DB00083 blocks neuropeptide release from afferent nerves , exocytosis of acetylcholine and purines from efferent nerves , and possibly DB00171 release from the urothelium . Suppression by DB00083 of the surface expression of nociceptor channels on bladder afferents might also contribute to its improvement of urological sensory symptoms . The role of the synaptic protein snap-25 in the potency of botulinum neurotoxin type A . Botulinum neurotoxin serotype A ( DB00083 ) is distinguished from BoNT/E by longer duration of paralysis and greater potency . The proteolytic activity of DB00083 in cultures of dissociated spinal cord neurons persists beyond 80 days , whereas BoNT/E activity persists for less than 1 day ( Keller , J. E. , Neale , E. A. Oyler , G. , and Adler , M. ( 1999 ) FEBS Lett. 456 , 137-142 ) . This single quality of toxin activity can account for the differences observed in the duration of muscle block . In the present work we sought to understand the basis for the apparent greater potency of DB00083 . BoNT/E cleaves a 26-amino acid fragment from the C terminus of the synaptic protein P60880 whereas DB00083 removes only nine residues creating a 197-amino acid fragment ( P197 ) that is 95 % the length of P60880 . We show that inhibition of neurotransmitter release by BoNT/E is equivalent to the damage caused to P60880 . However , synaptic blockade by DB00083 is greater than the extent of P60880 proteolysis . These findings can be explained if P197 produces an inhibitory effect on neurotransmitter release . A mathematical model of the experimentally determined relationship between P60880 damage and blockade of neurotransmission supports this interpretation . Furthermore , neurotransmitter release following complete cleavage of P60880 can be achieved by P197 , but with about 5-fold less sensitivity to external Ca(2+) . In this case , vesicular release is restored by increasing intracellular Ca(2+) . These data demonstrate that P197 competes with intact P60880 , but is unable to initiate normal synaptic vesicle fusion in physiological concentrations of Ca(2+) . The blockade of the neurotransmitter release apparatus by botulinum neurotoxins . The high toxicity of the seven serotypes of botulinum neurotoxins ( DB00083 to G ) , together with their specificity and reversibility , includes them in the list A of potential bioterrorism weapons and , at the same time , among the therapeutics of choice for a variety of human syndromes . They invade nerve terminals and cleave specifically the three proteins which form the heterotrimeric P60880 REceptors ( SNARE ) complex that mediates neurotransmitter release . The BoNT-induced cleavage of the SNARE proteins explains by itself the paralysing activity of the BoNTs because the truncated proteins can not form the SNARE complex . However , in the case of DB00083 , the most widely used toxin in therapy , additional factors come into play as it only removes a few residues from the synaptosomal associate protein of 25 kDa C-terminus and this results in a long duration of action . To explain these facts and other experimental data , we present here a model for the assembly of the neuroexocytosis apparatus in which Synaptotagmin and Complexin first assist the zippering of the SNARE complex , and then stabilize and clamp an octameric radial assembly of the SNARE complexes . Disruption of pancreatic beta-cell lipid rafts modifies Kv2.1 channel gating and insulin exocytosis . In pancreatic beta-cells , the predominant voltage-gated Ca(2+) channel ( Ca(V)1.2 ) and K(+) channel ( K(V)2.1 ) are directly coupled to SNARE ( soluble N-ethylmaleimide-sensitive factor attachment protein ( P60880 ) receptor ) proteins . These SNARE proteins modulate channel expression and gating and closely associate these channels with the insulin secretory vesicles . We show that K(V)2.1 and Ca(V)1.2 , but not K(V)1.4 , Q09428 , or Kir6.2 , target to specialized cholesterol-rich lipid raft domains on beta-cell plasma membranes . Similarly , the SNARE proteins syntaxin 1A , P60880 , and P63027 , but not Munc-13-1 or n-Sec1 , are associated with lipid rafts . Disruption of the lipid rafts by depleting membrane cholesterol with methyl-beta-cyclodextrin shunts K(V)2.1 , Ca(V)1.2 , and SNARE proteins out of lipid rafts . Furthermore , methyl-beta-cyclodextrin inhibits K(V)2.1 but not Ca(V)1.2 channel activity and enhances single-cell exocytic events and insulin secretion . Membrane compartmentalization of ion channels and SNARE proteins in lipid rafts may be critical for the temporal and spatial coordination of insulin release , forming what has been described as the excitosome complex . Self-assembled peptide monolayers as a toxin sensing mechanism within arrayed microchannels . A sensor for the lethal bacterial enzyme , botulinum neurotoxin type A ( DB00083 ) , was developed using self-assembled monolayers ( SAMs ) . SAMs consisting of an immobilized synthetic peptide that mimicked the toxin 's in vivo P60880 protein substrate were formed on Au and interfaced with arrayed microfluidic channels . Efforts to optimize DB00118 composition and assay conditions for greatest reaction efficiency and sensitivity are described in detail . Channel design provided facile fluid manipulation , sample incubation , analyte concentration , and fluorescence detection all within a single microfluidic channel , thus avoiding sample transfer and loss . Peptide SAMs were exposed to varying concentrations of DB00083 or its catalytic light chain ( ALC ) , resulting in enzymatic cleavage of the peptide substrate from the surface . Fluorescence detection was achieved down to 20 pg/mL ALC and 3 pg/mL DB00083 in 3 h . Toxin sensing was also accomplished in vegetable soup , demonstrating practicality of the method . The modular design of this microfluidic DB00118 platform allows for extension to sensing other toxins that operate via enzymatic cleavage , such as the remaining BoNT serotypes B-G , anthrax , and tetanus toxin . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Probing DB00083 protease exosites : implications for inhibitor design and light chain longevity . Botulinum neurotoxin serotype A ( DB00083 ) is one of the most lethal toxins known . Its extreme toxicity is due to its light chain ( LC ) , a zinc protease that cleaves P60880 , a synaptosome-associated protein , leading to the inhibition of neuronal activity . Studies on DB00083 LC have revealed that two regions , termed exosites , can play an important role in BoNT catalytic activity . A clear understanding of how these exosites influence neurotoxin catalytic activity would provide a critical framework for deciphering the mechanism of P60880 cleavage and the design of inhibitors . Herein , based on the crystallographic structure of DB00083 LC complexed with its substrate , we designed an α-exosite binding probe . Experiments with this unique probe demonstrated that α-exosite binding enhanced both catalytic activity and stability of the LC . These data help delineate why α-exosite binding is needed for P60880 cleavage and also provide new insights into the extended lifetime observed for DB00083 LC in vivo . Expression and purification of the light chain of botulinum neurotoxin A : a single mutation abolishes its cleavage of P60880 and neurotoxicity after reconstitution with the heavy chain . Botulinum neurotoxin type A ( DB00083 ) selectively and irreversibly inhibits acetylcholine release from peripheral nerve endings . While the toxin 's heavy ( H ) chain contributes to neuronal binding and internalization , its light ( L ) chain is a Zn(2+)-dependent endoprotease that intracellularly cleaves synaptosomal-associated protein of M(r) = 25 kDa ( P60880 ) . For research and clinical exploitation of this uniquely-acting neurotoxin , recombinant wild-type L chain was produced together with a mutant in which His227 in the Zn(2+)-binding motif was substituted by DB00135 . The PCR-amplified wild-type and mutant L chain genes were cloned , fused to the gene for maltose-binding protein , and expressed at high levels in Escherichia coli . The soluble fusion proteins were purified using amylose affinity chromatography , and , after factor Xa cleavage , the free L chains were isolated . The wild-type was shown to proteolyze P60880 at a rate approaching that of the native chain while the mutant was inactive . Reconstitution of the pure wild-type L chain with native homogeneous H chain yielded a disulfide-linked dichain form that inhibited neuromuscular transmission in vitro and produced the symptoms of botulism in vivo . After reconstitution with the H chain , the Tyr227 mutant L chain failed to show any neuroparalytic activity in either of these assays . This methodology allows , for the first time , routine preparation of recombinant forms of the L chain that are needed to decipher the molecular details of its interaction with substrate and , thereby , assist the design of effective inhibitors. ( ABSTRACT TRUNCATED AT 250 WORDS ) Primary cultures of embryonic chicken neurons for sensitive cell-based assay of botulinum neurotoxin : implications for therapeutic discovery . Botulinum toxin is an exceedingly potent inhibitor of neurotransmission across the neuromuscular junction , causing flaccid paralysis and death . The potential for misuse of this deadly poison as a bioweapon has added a greater urgency to the search for effective therapeutics . The development of sensitive and efficient cell-based assays for the evaluation of toxin antagonists is crucial to the rapid and successful identification of therapeutic compounds . The authors evaluated the sensitivity of primary cultures from 4 distinct regions of the embryonic chick nervous system to botulinum neurotoxin A ( DB00083 ) cleavage of synaptosomal-associated protein of 25 kD ( P60880 ) . Although differences in sensitivity were apparent , P60880 cleavage was detectable in neuronal cells from each of the 4 regions within 3 h at DB00083 concentrations of 1 nM or lower . Co-incubation of chick neurons with DB00083 and toxin-neutralizing antibodies inhibited P60880 cleavage , demonstrating the utility of these cultures for the assay of DB00083 antagonists . Basic tetrapeptides as potent intracellular inhibitors of type A botulinum neurotoxin protease activity . Botulinum neurotoxins ( BoNT ) are the most potent of all toxins that cause flaccid muscle paralysis leading to death . They are also potential biothreat agents . A systematic investigation of various short peptide inhibitors of the BoNT protease domain with a 17-residue peptide substrate led to arginine-arginine-glycine-cysteine having a basic tetrapeptide structure as the most potent inhibitor . When assayed in the presence of dithiothreitol ( DTT ) , the inhibitory effect was drastically reduced . Replacing the terminal cysteine with one hydrophobic residue eliminated the DTT effect but with two hydrophobic residues made the pentapeptide a poor inhibitor . Replacing the first arginine with cysteine or adding an additional cysteine at the N terminus did not improve inhibition . When assessed using mouse brain lysates , the tetrapeptides also inhibited DB00083 cleavage of the endogenous P60880 . The peptides penetrated the neuronal cell lines , N2A and BE(2)-M17 , without adversely affecting metabolic functions as measured by DB00171 production and P-38 phosphorylation . Biological activity of the peptides persisted within cultured chick motor neurons and rat and mouse cerebellar neurons for more than 40 h and inhibited DB00083 protease action inside the neurons in a dose- and time-dependent fashion . Our results define a tetrapeptide as the smallest peptide inhibitor in the backdrop of a large substrate protein of 200+ amino acids having multiple interaction regions with its cognate enzyme . The inhibitors should also be valuable candidates for drug development . Comparative in vitro effects of guinea pig P01282 and common P01282 on liver and lung membranes from guinea pig and rat and on human lymphoblastic P60880 -T1 membranes . Guinea pig P01282 differs from P01282 of several mammals by its amino acids in positions 5 , 9 , 19 and 26 . We tested a ) its ability to occupy P01282 receptors in liver and lung membranes of rat and guinea pig and in the human lymphoblastic P60880 -T1 cell line and b ) the ensuing adenylate cyclase stimulation . In liver and lung membranes from rat , guinea pig P01282 was less potent than common P01282 to occupy high and low affinity P01282 receptors . In rat liver both P01282 activated adenylate cyclase mostly through high affinity receptors . In rat lung , guinea pig P01282 activated the enzyme mostly through high affinity receptors and was less efficient than common P01282 acting through both classes of receptors . In guinea pig liver and lung membranes , binding inhibition curves were steeper than with rat preparations and adenylate cyclase appeared to be mostly activated through high affinity P01282 receptors in liver and through both classes of receptors in lung . On human lymphoblastic P60880 -T1 membranes both P01282 were equally potent and efficient to inhibit tracer binding and activate adenylate cyclase . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Light chain of botulinum A neurotoxin expressed as an inclusion body from a synthetic gene is catalytically and functionally active . Botulinum neurotoxins , the most potent of all toxins , induce lethal neuromuscular paralysis by inhibiting exocytosis at the neuromuscular junction . The light chains ( LC ) of these dichain neurotoxins are a new class of zinc-endopeptidases that specifically cleave the synaptosomal proteins , P60880 , VAMP , or syntaxin at discrete sites . To facilitate the structural and functional characterization of these unique endopeptidases , we constructed a synthetic gene for the LC of the botulinum neurotoxin serotype A ( DB00083 ) , overexpressed it in Escherichia coli , and purified the gene product from inclusion bodies . Our procedure can provide 1.1 g of the LC from 1 L of culture . The LC product was stable in solution at 4 degrees C for at least 6 months . This rBoNT/A LC was proteolytically active , specifically cleaving the DB00142 - DB00125 bond in a 17-residue synthetic peptide of P60880 , the reported cleavage site of DB00083 . Its calculated catalytic efficiency kcat/Km was higher than that reported for the native DB00083 dichain . Treating the rBoNT/A LC with mercuric compounds completely abolished its activity , most probably by modifying the cysteine-164 residue located in the vicinity of the active site . About 70 % activity of the LC was restored by adding DB01593 to a DB01593 -free , apo-LC preparation . The LC was nontoxic to mice and failed to elicit neutralizing epitope(s) when the animals were vaccinated with this protein . In addition , injecting rBoNT/A LC into sea urchin eggs inhibited exocytosis-dependent plasma membrane resealing . For the first time , results of our study make available a large amount of the biologically active toxin fragment in a soluble and stable form . Novel chimeras of botulinum neurotoxins A and E unveil contributions from the binding , translocation , and protease domains to their functional characteristics . Hyperexcitability disorders of cholinergically innervated muscles are treatable with botulinum neurotoxin ( BoNT ) A . The seven serotypes ( A-G ) potently block neurotransmission by binding to presynaptic receptors , undergoing endocytosis , transferring to the cytosol , and inactivating proteins essential for vesicle fusion . Although DB00083 and BoNT/E cleave P60880 , albeit at distinct sites , BoNT/E blocks neurotransmission faster and more potently . To identify the domains responsible for these characteristics , the C-terminal heavy chain portions of DB00083 and BoNT/E were exchanged to create chimeras AE and EA . After high yield expression in Escherichia coli , these single chain chimeras were purified by two-step chromatography and activated by conversion to disulfide-linked dichains . In vitro , each entered neurons , cleaved P60880 , and blocked neuromuscular transmission while causing flaccid paralysis in vivo . Acidification-dependent translocation of the light chain to the cytosol occurred more rapidly for BoNT/E and EA than for DB00083 and AE because the latter pair remained susceptible for longer to inhibitors of the vesicular proton pump , and DB00083 proved less sensitive . The receptor-binding and protease domains do not seem to be responsible for the speeds of intoxication ; rather the N-terminal halves of their heavy chains are implicated , with dissimilar rates of cytosolic transfer of the light chains being due to differences in pH sensitivity . AE produced the most persistent muscle weakening and therefore has therapeutic potential . Thus , proof of principle is provided for tailoring the pharmacological properties of these toxins by protein engineering . Domain requirement for the membrane trafficking and targeting of syntaxin 1A . Syntaxin plays a key role in intracellular membrane fusion in eukaryotic cells . The function of syntaxin relies on its proper trafficking to and targeting at the target membrane . The mechanisms underlying the trafficking and targeting of syntaxin to its physiological sites remain poorly understood . Here we have analyzed the trafficking of syntaxin 1A in P01308 -1 and CHO cells . We have identified the transmembrane domain together with several flanking positive-charged amino acids as the minimal domain required for the membrane delivery . Interestingly , we found that SNARE motif-exposed syntaxin 1A mutants were retained in endoplasmic reticulum ( ER ) and failed to transport to the cell surface in the absence of P60880 , suggesting that the exposure of the SNARE motif causes ER retention and complexation with P60880 helps the ER escape . Finally , our data propose two key roles for the H(abc) domain : to protect nonspecific interaction by masking the SNARE motif and to participate in the clustering of syntaxin 1A to the fusion sites in the plasma membrane . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Ca2+ or Sr2+ partially rescues synaptic transmission in hippocampal cultures treated with botulinum toxin A and C , but not tetanus toxin . Botulinum ( DB00083 -G ) and tetanus toxins ( TeNT ) are zinc endopeptidases that cleave proteins associated with presynaptic terminals ( P60880 , syntaxin , or VAMP/synaptobrevin ) and block neurotransmitter release . Treatment of hippocampal slice cultures with DB00083 , BoNT/C , BoNT/E , or TeNT prevented the occurrence of spontaneous or miniature EPSCs ( sEPSCs or mEPSCs ) as well as the [Ca2+]o-independent increase in their frequency induced by phorbol ester , 0.5 nM alpha-latrotoxin , or sucrose . [Ca2+]o-independent and -dependent release thus requires that the target proteins of clostridial neurotoxins be uncleaved . In contrast , significant increases in mEPSC frequency were produced in BoNT-treated , but not TeNT-treated , cultures by application of the Ca2+ ionophore ionomycin in the presence of 10 mM [Ca2+]o . The frequency of sEPSCs was increased in BoNT-treated , but not TeNT-treated , cultures by increasing [Ca2+]o from 2.8 to 5-10 mM or by applying 5 mM Sr2+ . Large Ca2+ and Sr2+ influxes thus can rescue release after BoNT treatment , albeit less than in control cultures . The nature of the toxin-induced modification of Ca2+-dependent release was assessed by recordings from monosynaptically coupled P07451 cell pairs . The paired-pulse ratio of unitary EPSCs evoked by two presynaptic action potentials in close succession was 0.5 in control cultures , but it was 1.4 and 1.2 in DB00083 - or BoNT/C-treated cultures when recorded in 10 mM [Ca2+]o . Log-log plots of unitary EPSC amplitude versus [Ca2+]o were shifted toward higher [Ca2+]o in DB00083 - or BoNT/C-treated cultures , but their slope was unchanged and the maximal EPSC amplitudes were reduced . We conclude that BoNTs reduce the Ca2+ sensitivity of the exocytotic machinery and the number of quanta released . Embryonic stem cell-derived neurons are a novel , highly sensitive tissue culture platform for botulinum research . There are no pharmacological treatments to rescue botulinum neurotoxin ( BoNT ) -mediated paralysis of neuromuscular signaling . In part , this failure can be attributed to the lack of a cell culture model system that is neuron-based , allowing detailed elucidation of the mechanisms underlying BoNT pathogenesis , yet still compatible with modern cellular and molecular approaches . We have developed a method to derive highly enriched , glutamatergic neurons from suspension-cultured murine embryonic stem ( ES ) cells . Hypothesizing that ES cell-derived neurons ( ESNs ) might comprise a novel platform to investigate the neurotoxicology of BoNTs , we evaluated the susceptibility of ESNs to DB00083 and BoNT/E using molecular and functional assays . ESNs express neuron-specific proteins , develop synapses and release glutamate in a calcium-dependent manner under depolarizing conditions . They express the BoNT substrate SNARE proteins P60880 , P63027 and syntaxin , and treatment with DB00083 and BoNT/E holotoxin results in proteolysis of P60880 within 24 h with EC50s of 0.81 and 68.6 pM , respectively . Intoxication with DB00083 results in the functional inhibition of potassium-induced , calcium-dependent glutamate release . ESNs remain viable and susceptible to intoxication for up to 90 days after plating , enabling longitudinal screens exploring toxin-specific mechanisms underlying persistence of synaptic blockade . The evidence suggests that derived neurons are a novel , biologically relevant model system that combines the verisimilitude of primary neurons with the genetic tractability and scalable expansion of a continuous cell line , and thus should significantly accelerate BoNT research and drug discovery while dramatically decreasing animal use . Recombinant P60880 is an effective substrate for Clostridium botulinum type A toxin endopeptidase activity in vitro . Bacterial neurotoxins are now being used routinely for the treatment of neuromuscular conditions . Alternative assays to replace or to complement in vivo bioassay methods for assessment of the safety and potency of these botulinum neurotoxin-based therapeutic products are urgently needed . Advances made in understanding the mode of action of clostridial neurotoxins have provided the basis for the development of alternative mechanism-based assay methods . Thus , the identification of P60880 ( synaptosomal-associated protein of molecular mass 25 kDa ) as the intracellular protein target which is selectively cleaved during poisoning by botulinum neurotoxin type A ( DB00083 ) has enabled the development of a functional in vitro assay for this toxin . Using recombinant DNA methods , a segment of P60880 ( aa residues 134-206 ) spanning the toxin cleavage site was prepared as a fusion protein to the maltose-binding protein in Escherichia coli . The fusion protein was purified by affinity chromatography and the fragment isolated after cleavage with Factor Xa . Targeted antibodies specific for the N and C termini of P60880 , as well as the toxin cleavage site , were prepared and used in an immunoassay to demonstrate DB00083 endopeptidase activity towards recombinant P60880 substrates . The reaction required low concentrations of reducing agents which were inhibitory at higher concentrations as were metal chelators and some inhibitors of metallopeptidases . The endopeptidase assay has proved to be more sensitive than the mouse bioassay for detection of toxin in therapeutic preparations . A good correlation with results obtained in the in vivo bioassay ( r = 0.95 , n = 23 ) was demonstrated . The endopeptidase assay described here may provide a suitable replacement assay for the estimation of the potency of type A toxin in therapeutic preparations . Activation of Q8NER1 mediates calcitonin gene-related peptide release , which excites trigeminal sensory neurons and is attenuated by a retargeted botulinum toxin with anti-nociceptive potential . Excessive release of inflammatory/pain mediators from peripheral sensory afferents renders nerve endings hyper-responsive , causing central sensitization and chronic pain . Herein , the basal release of proinflammatory calcitonin gene-related peptide ( P80511 ) was shown to increase the excitability of trigeminal sensory neurons in brainstem slices via CGRP1 receptors because the effect was negated by an antagonist , CGRP8-37 . This excitatory action could be prevented by cleaving synaptosomal-associated protein of M(r) 25,000 ( P60880 ) with botulinum neurotoxin ( BoNT ) type A , a potent inhibitor of exocytosis . Strikingly , DB00083 proved unable to abolish the CGRP1 receptor-mediated effect of capsaicin , a nociceptive Q8NER1 stimulant , or its elevation of P80511 release from trigeminal ganglionic neurons ( TGNs ) in culture . Although the latter was also not susceptible to BoNT/E , apparently attributable to a paucity of its acceptors ( glycosylated synaptic vesicle protein 2 A/B ) , this was overcome by using a recombinant chimera ( EA ) of DB00083 and BoNT/E . It bound effectively to the C isoform of SV2 abundantly expressed in TGNs and cleaved P60880 , indicating that its /A binding domain ( H(C) ) mediated uptake of the active /E protease . The efficacy of /EA is attributable to removal of 26 C-terminal residues from P60880 , precluding formation of SDS-resistant SNARE complexes . In contrast , exocytosis could be evoked after deleting nine of the P60880 residues with /A but only on prolonged elevation of [Ca(2+)](i) with capsaicin . This successful targeting of /EA to nociceptive neurons and inhibition of P80511 release in vitro and in situ highlight its potential as a new therapy for sensory dysmodulation and chronic pain . Neuronal targeting , internalization , and biological activity of a recombinant atoxic derivative of botulinum neurotoxin A . Non-toxic derivatives of botulinum neurotoxin A ( DB00083 ) have potential use as neuron-targeting delivery vehicles , and as reagents to study intracellular trafficking . We have designed and expressed an atoxic derivative of DB00083 ( DB00083 ad ) as a full-length 150 kDa molecule consisting of a 50 kDa light chain ( LC ) and a 100 kDa heavy chain ( HC ) joined by a disulfide bond and rendered atoxic through the introduction of metalloprotease-inactivating point mutations in the light chain . Studies in neuronal cultures demonstrated that DB00083 ad can not cleave synaptosomal-associated protein 25 ( P60880 ) , the substrate of wt DB00083 , and that it effectively competes with wt DB00083 for binding to endogenous neuronal receptors . In vitro and in vivo studies indicate accumulation of DB00083 ad at the neuromuscular junction of the mouse diaphragm . Immunoprecipitation studies indicate that the LC of DB00083 ad forms a complex with P60880 present in the neuronal cytosolic fraction , demonstrating that the atoxic LC retains the P60880 binding capability of the wt toxin . Toxicity of DB00083 ad was found to be reduced approximately 100,000-fold relative to wt DB00083 . Characterization of Förster resonance energy transfer in a botulinum neurotoxin protease assay . Our previous article described a fluorescence-based assay for monitoring the proteolytic activity of botulinum neurotoxin types A and E ( DB00083 and BoNT/E ) . As detailed in that article , the assay is based on depolarization due to Förster resonance energy transfer between blue fluorescent protein ( Q9ULX5 ) and green fluorescent protein ( GFP ) moieties linked via residues 134-206 of P60880 ( synaptosome-associated protein of 25kDa ) , the protein substrate for DB00083 and BoNT/E . Before cleavage of this recombinant substrate , the polarization observed for the GFP emission , excited near the absorption maximum of the Q9ULX5 , is very low due to depolarization following energy transfer from Q9ULX5 to GFP . After substrate cleavage and diffusion of the fluorescent proteins beyond the energy transfer distance , the polarization is high due to observation of the emission only from directly excited GFP . This change in fluorescence polarization allows an assay , termed DARET ( depolarization after resonance energy transfer ) , that is robust and sensitive . In this article , we characterize the spectroscopic parameters of the system before and after substrate cleavage , including excitation and emission spectra , polarizations , and lifetimes . Anti-nociceptive effect of a conjugate of DB05875 and light chain of botulinum neurotoxin type A . Neuropathic pain is a debilitating condition resulting from damage to sensory transmission pathways in the peripheral and central nervous system . A potential new way of treating chronic neuropathic pain is to target specific pain-processing neurons based on their expression of particular receptor molecules . We hypothesized that a toxin-neuropeptide conjugate would alter pain by first being taken up by specific receptors for the neuropeptide expressed on the neuronal cells . Then , once inside the cell the toxin would inhibit the neurons ' activity without killing the neurons , thereby providing pain relief without lesioning the nervous system . In an effort to inactivate the nociceptive neurons in the trigeminal nucleus caudalis in mice , we targeted the NK1 receptor ( P25103 ) using DB05875 ( SP ) . The catalytically active light chain of botulinum neurotoxin type A ( LC/A ) was conjugated with SP . Our results indicate that the conjugate DB00083 -LC:SP is internalized in cultured P25103 -expressing neurons and also cleaves the target of botulinum toxin , a component-docking motif necessary for release of neurotransmitters called P60880 . The conjugate was next tested in a murine model of DB01229 -induced neuropathic pain . An intracisternal injection of DB00083 -LC:SP decreased thermal hyperalgesia as measured by the operant orofacial nociception assay . These findings indicate that conjugates of the light chain of botulinum toxin are extremely promising agents for use in suppressing neuronal activity for extended time periods , and that DB00083 -LC:SP may be a useful agent for treating chronic pain . Widespread sequence variations in P23763 across vertebrates suggest a potential selective pressure from botulinum neurotoxins . Botulinum neurotoxins ( DB00083 -G ) , the most potent toxins known , act by cleaving three SNARE proteins required for synaptic vesicle exocytosis . Previous studies on BoNTs have generally utilized the major SNARE homologues expressed in brain ( P63027 , syntaxin 1 , and P60880 ) . However , BoNTs target peripheral motor neurons and cause death by paralyzing respiratory muscles such as the diaphragm . Here we report that P23763 , but not P63027 , is the SNARE homologue predominantly expressed in adult rodent diaphragm motor nerve terminals and in differentiated human motor neurons . In contrast to the highly conserved P63027 , BoNT-resistant variations in P23763 are widespread across vertebrates . In particular , we identified a polymorphism at position 48 of P23763 in rats , which renders P23763 either resistant ( I48 ) or sensitive ( M48 ) to BoNT/D . Taking advantage of this finding , we showed that rat diaphragms with I48 in P23763 are insensitive to BoNT/D compared to rat diaphragms with M48 in P23763 . This unique intra-species comparison establishes P23763 as a physiological toxin target in diaphragm motor nerve terminals , and demonstrates that the resistance of P23763 to BoNTs can underlie the insensitivity of a species to members of BoNTs . Consistently , human P23763 contains I48 , which may explain why humans are insensitive to BoNT/D . Finally , we report that residue 48 of P23763 varies frequently between M and I across seventeen closely related primate species , suggesting a potential selective pressure from members of BoNTs for resistance in vertebrates . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Persistence of botulinum neurotoxin action in cultured spinal cord cells . Primary dissociated fetal mouse spinal cord cultures were used to study the mechanisms underlying the differences in persistence of botulinum neurotoxin A ( DB00083 ) and botulinum neurotoxin/E ( BoNT/E ) activities . Spinal cord cultures were exposed to DB00083 ( 0.4 pM ) for 2-3 days , which converted approximately half of the P60880 to an altered form lacking the final nine C-terminal residues . The distribution of toxin-damaged to control P60880 remained relatively unchanged for up to 80 days thereafter . Application of a high concentration of BoNT/E ( 250 pM ) either 25 or 60 days following initial intoxication with DB00083 converted both normal and DB00083 -truncated P60880 into a single population lacking the final 26 C-terminal residues . Excess BoNT/E was removed by washout , and recovery of intact P60880 was monitored by Western blot analysis . The BoNT/E-truncated species gradually diminished during the ensuing 18 days , accompanied by the reappearance of both normal and DB00083 -truncated P60880 . Return of DB00083 -truncated P60880 was observed in spite of the absence of DB00083 in the culture medium during all but the first 3 days of exposure . These results indicate that proteolytic activity associated with the DB00083 light chain persists inside cells for > 11 weeks , while recovery from BoNT/E is complete in < 3 weeks . This longer duration of enzymatic activity appears to account for the persistence of serotype A action . Proteolysis of P60880 by types E and A botulinal neurotoxins . Clostridial neurotoxins , tetanus toxin ( TeTx ) and the seven related but serologically distinct botulinal neurotoxins ( DB00083 to BoNT/G ) , are potent inhibitors of synaptic vesicle exocytosis in nerve endings . Recently it was reported that the light chains of clostridial neurotoxins act as zinc-dependent metalloproteases which specifically cleave synaptic target proteins such as synaptobrevin/VAMPs , HPC-1/syntaxin ( BoNT/C1 ) , and P60880 ( DB00083 ) . We show here that BoNT/E , like DB00083 , cleaves P60880 , as generated by in vitro translation or by expression in Escherichia coli . BoNT/E cleaves the Arg180-Ile181 bond . This site is different from that of DB00083 , which cleaves P60880 between the amino acid residues Gln197 and Arg198 . These findings further support the view that clostridial neurotoxins have evolved from an ancestral protease recognizing the exocytotic fusion machinery of synaptic vesicles whereby individual toxins target different members of the membrane fusion complex . DB00368 inhibits exocytosis via the G protein βγ subunit and refilling of the readily releasable granule pool via the α(i1/2) subunit . The molecular mechanisms responsible for the ' distal ' effect by which noradrenaline ( NA ) blocks exocytosis in the β-cell were examined by whole-cell and cell-attached patch clamp capacitance measurements in P01308 832/13 β-cells . NA inhibited Ca(2+)-evoked exocytosis by reducing the number of exocytotic events , without modifying vesicle size . Fusion pore properties also were unaffected . NA-induced inhibition of exocytosis was abolished by a high level of Ca(2+) influx , by intracellular application of antibodies against the G protein subunit Gβ and was mimicked by the myristoylated βγ-binding/activating peptide mSIRK . NA-induced inhibition was also abolished by treatment with DB00083 , which cleaves the C-terminal nine amino acids of P60880 , and also by a P60880 C-terminal-blocking peptide containing the DB00083 cleavage site . These data indicate that inhibition of exocytosis by NA is downstream of increased [Ca(2+)](i) and is mediated by an interaction between Gβγ and the C-terminus of P60880 , as is the case for inhibition of neurotransmitter release . Remarkably , in the course of this work , a novel effect of NA was discovered . NA induced a marked retardation of the rate of refilling of the readily releasable pool ( O75783 ) of secretory granules . This retardation was specifically abolished by a Gα(i1/2) blocking peptide demonstrating that the effect is mediated via activation of Gα(i1) and/or Gα(i2) . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Association of botulinum neurotoxin serotype A light chain with plasma membrane-bound P60880 . The Clostridium botulinum neurotoxins ( BoNTs ) cleave SNARE proteins , which inhibit binding and thus fusion of neurotransmitter vesicles to the plasma membrane of peripheral neurons . BoNTs comprise an N-terminal light chain ( LC ) and C-terminal heavy chain , which are linked by a disulfide bond . There are seven serotypes ( A-G ) of BoNTs based upon immunological neutralization . Although the binding and entry of DB00083 into neurons has been subjected to considerable investigation , the intracellular events that allow DB00083 to efficiently cleave P60880 within neurons is less well understood . Earlier studies showed that intracellular LC/A bound to the plasma membrane of neurons . In this study , intracellular LC/A is shown to directly bind P60880 on the plasma membrane . Solid phase binding showed that the N-terminal residues of LC/A bound residues 80-110 of P60880 , which was also observed in cultured neurons . Association of the N-terminal 8 amino acids of LC/A and residues 80-110 of P60880 also enhanced substrate cleavage . These findings explain how LC/A associates with P60880 on the plasma membrane and provide a basis for LC/A cleavage of P60880 within the SNARE complex . Structural analysis of botulinum neurotoxin serotype F light chain : implications on substrate binding and inhibitor design . The seven serologically distinct Clostridium botulinum neurotoxins ( BoNTs A-G ) are zinc endopeptidases which block the neurotransmitter release by cleaving one of the three proteins of the soluble N-ethylmaleimide-sensitive-factor attachment protein receptor complex ( SNARE complex ) essential for the fusion of vesicles containing neurotransmitters with target membranes . These metallopeptidases exhibit unique specificity for the substrates and peptide bonds they cleave . Development of countermeasures and therapeutics for BoNTs is a priority because of their extreme toxicity and potential misuse as biowarfare agents . Though they share sequence homology and structural similarity , the structural information on each one of them is required to understand the mechanism of action of all of them because of their specificity . Unraveling the mechanism will help in the ultimate goal of developing inhibitors as antibotulinum drugs for the toxins . Here , we report the high-resolution structure of active BoNT/F catalytic domain in two crystal forms . The structure was exploited for modeling the substrate binding and identifying the S1 ' subsite and the putative exosites which are different from DB00083 or BoNT/B . The orientation of docking of the substrate at the active site is consistent with the experimental DB00083 -LC: P60880 peptide model and our proposed model for BoNT/E-LC: P60880 . Botulinum neurotoxin type A : Actions beyond P60880 ? Botulinum neurotoxin type A ( DB00083 ) , the most potent toxin known in nature which causes botulism , is a commonly used therapeutic protein . It prevents synaptic vesicle neuroexocytosis by proteolytic cleavage of synaptosomal-associated protein of 25 kDa ( P60880 ) . It is widely believed that DB00083 therapeutic or toxic actions are exclusively mediated by P60880 cleavage . On the other hand , in vitro and in vivo findings suggest that several DB00083 actions related to neuroexocytosis , cell cycle and apoptosis , neuritogenesis and gene expression are not necessarily mediated by this widely accepted mechanism of action . In present review we summarize the literature evidence which point to the existence of unknown DB00083 molecular target(s) and modulation of unknown signaling pathways . The effects of DB00083 apparently independent of P60880 occur at similar doses/concentrations known to induce P60880 cleavage and prevention of neurotransmitter release . Accordingly , these effects might be pharmacologically significant . Potentially the most interesting are observations of antimitotic and antitumor activity of DB00083 . However , the exact mechanisms require further studies . Truncation of P60880 reduces the stimulatory action of DB02527 on rapid exocytosis in insulin-secreting cells . Synaptosomal protein of 25 kDa ( P60880 ) is important for Ca(2+)-dependent fusion of large dense core vesicles ( LDCVs ) in insulin-secreting cells . Exocytosis is further enhanced by DB02527 -increasing agents such as glucagon-like peptide-1 ( P0C6A0 ) , and this augmentation includes interaction with both PKA and Q8WZA2 . To investigate the coupling between P60880 - and DB02527 -dependent stimulation of insulin exocytosis , we have used capacitance measurements , protein-binding assays , and Western blot analysis . In insulin-secreting P01308 -1 cells overexpressing wild-type P60880 ( P60880 (WT) ) , rapid exocytosis was stimulated more than threefold by DB02527 , similar to the situation in nontransfected cells . However , DB02527 failed to potentiate rapid exocytosis in P01308 -1 cells overexpressing a truncated form of P60880 ( P60880 (1-197) ) or Botulinum neurotoxin A ( DB00083 ) . Close dissection of the exocytotic response revealed that the inability of DB02527 to stimulate exocytosis in the presence of a truncated P60880 was confined to the release of primed LDCVs within the readily releasable pool , especially from the immediately releasable pool , whereas DB02527 enhanced mobilization of granules from the reserve pool in both P60880 (1-197) ( P < 0.01 ) and P60880 (WT) ( P < 0.05 ) cells . This was supported by hormone release measurements . Augmentation of the immediately releasable pool by DB02527 has been suggested to act through the Q8WZA2 -dependent , PKA-independent pathway . Indeed , we were able to verify an interaction between P60880 with both Q8WZA2 and Q9UQ26 , two proteins involved in the PKA-independent pathway . Thus we hypothesize that P60880 is a necessary partner in the complex mediating DB02527 -enhanced rapid exocytosis in insulin-secreting cells . Stable expression of a neuronal dopaminergic progenitor phenotype in cell lines derived from human amniotic fluid cells . Cells from human amniotic fluid derived from the fetus are considered a source of multipotent cells . Their properties have not been fully exploited , partially because unlike other embryonic sources such as embryonic stem ( ES ) cells , cell lines from amniocentesis samples have not been generated . We have established and characterized the properties of eight individual cell lines . Flow cytometry using several cell surface markers showed that all cell lines generated consisted of homogeneous populations that lack HLAII antigenicity . Using a combination of immunocytochemistry , Western blotting , and RT-PCR , we found weak expression of Oct4 and nestin and strong expression of tubulin-betaIII , P11137 , and tau . Specific markers for cholinergic , (nor)adrenergic , and GABAergic neurons or glia were weakly expressed or absent , whereas expression of factors implicated in early induction of dopaminergic neurons , TGF-beta3 and beta-catenin were present . Further analysis showed strong expression of EN-1 , c- P07949 , PTX3 , and P43354 essential for induction and survival of midbrain dopaminergic neurons , TH , P20711 , and Q05940 components of dopamine synthesis and secretion , and syntaxin1A and P60880 necessary for neurotransmitter exocytosis . This phenotype was retained throughout passages and up to the current passage 36 . Expression of neuronal and dopaminergic markers in individual AF cell lines was comparable to expression in neurons induced from ES cells and in IMR-32 and SH-SY5Y neuroblastomas . Our data show that cell lines can be derived from subcultures of amniocentesis , and are primarily composed of a population of progenitors with a phenotype similar to that of committed mesencephalic dopaminergic neurons . 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 . Inhibition of interleukin-12 p40 transcription and NF-kappaB activation by nitric oxide in murine macrophages and dendritic cells . DB00435 ( NO ) , an important effector molecule of the innate immune system , can also regulate adaptive immunity . In this study , the molecular effects of NO on the toll-like receptor signaling pathway were determined using interleukin-12 ( IL-12 ) as an immunologically relevant target gene . The principal conclusion of these experiments is that NO inhibits IL-1 receptor-associated kinase ( P51617 ) activity and attenuates the molecular interaction between tumor necrosis factor receptor-associated factor-6 and P51617 . As a consequence , the NO donor S-nitroso-N-acetylpenicillamine ( P60880 ) inhibits lipopolysaccharide ( LPS ) -induced IL-12 p40 mRNA expression , protein production , and promoter activity in murine macrophages , dendritic cells , and the murine macrophage cell line RAW 264.7 . Splenocytes from inducible nitric-oxide synthase-deficient mice demonstrate markedly increased IL-12 p40 protein and mRNA expression compared with wild type splenocytes . The inhibitory action of NO on IL-12 p40 is independent of the cytokine P22301 . The effects of NO can be directly attributed to inhibition of NF-kappaB activation through P51617 -dependent pathways . Accordingly , P60880 strongly reduces LPS-induced NF-kappaB DNA binding to the p40 promoter and inhibits LPS-induced IkappaB phosphorylation . Similarly , NO attenuates IL-1beta-induced NF-kappaB activation . These experiments provide another example of how an innate immune molecule may have a profound effect on adaptive immunity . Direct biosensor detection of botulinum neurotoxin endopeptidase activity in sera from patients with type A botulism . Botulinum neurotoxin A ( DB00083 ) has intrinsic endoprotease activity specific for P60880 , a key protein for presynaptic neurotransmitter release . The inactivation of P60880 by DB00083 underlies botulism , a rare but potentially fatal disease . There is a crucial need for a rapid and sensitive in vitro serological test for DB00083 to replace the current in vivo mouse bioassay . Cleavage of P60880 by DB00083 generates neo-epitopes which can be detected by binding of a monoclonal antibody ( mAb10F12 ) and thus measured by surface plasmon resonance ( SPR ) . We have explored two SPR assay formats , with either mAb10F12 or His6- P60880 coupled to the biosensor chip . When DB00083 was incubated with P60880 in solution and the reaction products were captured on a mAb-coated chip , a sensitivity of 5 fM ( 0.1LD50/ml serum ) was obtained . However , this configuration required prior immunoprecipitation of DB00083 . A sensitivity of 0.5 fM in 10 % serum ( 0.1 LD50/ml serum ) was attained when P60880 was coupled directly to the chip , followed by sequential injection of DB00083 samples and mAb10F12 into the flow system to achieve on-chip cleavage and detection , respectively . This latter format detected DB00083 endoprotease activity in 50-100 µl serum samples from all patients ( 11/11 ) with type A botulism within 5h . No false positives occurred in sera from healthy subjects or patients with other neurological diseases . The automated chip-based procedure has excellent specificity and sensitivity , with significant advantages over the mouse bioassay in terms of rapidity , required sample volume and animal ethics . Hypersensitive detection and quantitation of DB00083 by IgY antibody against substrate linear-peptide . Botulinum neurotoxin A ( DB00083 ) , the most acutely poisonous substance to humans known , cleave its P60880 substrate with high specificity . Based on the endopeptidase activity , different methods have been developed to detect DB00083 , but most lack ideal reproducibility or sensitivity , or suffer from long-term or unwanted interferences . In this study , we developed a simple method to detect and quantitate trace amounts of botulinum neurotoxin A using the IgY antibody against a linear-peptide substrate . The effects of reaction buffer , time , and temperature were analyzed and optimized . When the optimized assay was used to detect DB00083 , the limit of detection of the assay was 0.01 mouse LD50 ( 0.04 pg ) , and the limit of quantitation was 0.12 mouse LD50/ml ( 0.48 pg ) . The findings also showed favorable specificity of detecting DB00083 . When used to detect DB00083 in milk or human serum , the proposed assay exhibited good quantitative accuracy ( 88 % < recovery < 111 % ; inter- and intra-assay CVs < 18 % ) . This method of detection took less than 3 h to complete , indicating that it can be a valuable method of detecting DB00083 in food or clinical diagnosis . Neuromuscular transmission and muscle contractility in P60880 -deficient coloboma mice . Synaptosomal associated protein of 25 kDa ( P60880 ) is a cytoplasmic protein that participates in the docking and fusion of synaptic vesicles with the nerve terminal in preparation for neurotransmitter release . P60880 is also a substrate for three of the seven serotypes of botulinum neurotoxin ( BoNT ) . Intoxication by DB00083 , /C1 or /E results in weakness and paralysis of skeletal muscle due to cleavage of P60880 ( and syntaxin la in the case /C1 ) at discrete serotype-specific sites . To elucidate the role of P60880 in muscle function in more detail , contractility and neuromuscular transmission were studied in a mutant mouse model termed coloboma . The coloboma mutation results from a contiguous deletion of 1-2 centiMorgans on chromosome 2 , which includes the entire P60880 locus and three other identified genes . Homozygotes do not survive beyond gestation day 6 ; heterozygotes ( Cm/+ ) have a normal life-span but express reduced levels of P60880 mRNA and protein in the brain . The consequences of the Cm/+ mutation on twitch and tetanic tension , quantal release of neurotransmitter and spinal motoneuron expression of P60880 were examined in the present study . Contrary to expectations , Cm/+ mice exhibited no alteration in twitch tension and generated normal tetanic tension even at the highest frequency examined ( 800 Hz ) . Microelectrode recordings revealed that MEPP amplitude and frequency were both within control limits . The ventral spinal cord of Cm/+ mice showed no deficiency in P60880 content and immunohistochemical examination of nerve terminals in Cm/+ mice disclosed that P60880 levels and distribution were similar to those of control mice . It is concluded that spinal motor neurons up-regulate P60880 to preserve vital neuromuscular function . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . The role of nitric oxide in ischaemia/reperfusion injury of isolated hearts from severely atherosclerotic mice . DB00435 ( NO ) may play an essential role for maintenance of cardiac function and perfusion , while endothelial dysfunction of atherosclerotic vessels may aggravate ischaemia/reperfusion injury . This paper investigates the role of nitric oxide in ischaemia/reperfusion injury in hearts with coronary atherosclerosis . Hearts of apolipoprotein E/ P01130 double knockout ( ApoE/LDLr KO ) mice fed an atherogenic diet for 7-9 months were isolated and Langendorff-perfused with 40 minutes of global ischaemia and 60 minutes reperfusion , and funtion and infarction compared with hearts of C57BL/6 controls in the prescence or abscence of the NO-donor P60880 or the NOS inhibitor L-NAME . Hearts of animals with atherosclerosis were more susceptible to ischaemia/reperfusion injury than hearts of animals with healthy vessels , evident as more impaired left ventricular performance . P60880 protected function and reduced infarct size in atherosclerotic hearts , but the same concentration of P60880 was detrimental in normal hearts , perhaps due to NO-overproduction and peroxynitrite formation demonstrated immunohistochemically as increased formation of nitrosylated tyrosine . A low concentration of P60880 protected against ischaemia/reperfusion dysfunction in normal hearts . L-NAME decreased left ventricular performance in atherosclerotic hearts . These findings suggest that impaired endothelium dependent function contributes to reperfusion injury in coronary atherosclerosis . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Candidate gene studies of ADHD : a meta-analytic review . Quantitative genetic studies ( i.e. , twin and adoption studies ) suggest that genetic influences contribute substantially to the development of attention deficit hyperactivity disorder ( ADHD ) . Over the past 15 years , considerable efforts have been made to identify genes involved in the etiology of this disorder resulting in a large and often conflicting literature of candidate gene associations for ADHD . The first aim of the present study was to conduct a comprehensive meta-analytic review of this literature to determine which candidate genes show consistent evidence of association with childhood ADHD across studies . The second aim was to test for heterogeneity across studies in the effect sizes for each candidate gene as its presence might suggest moderating variables that could explain inconsistent results . Significant associations were identified for several candidate genes including Q01959 , P21917 , P21918 , P31645 , P28222 , and P60880 . Further , significant heterogeneity was observed for the associations between ADHD and Q01959 , P21917 , P21918 , P09172 , P08913 , P31645 , Q8IWU9 , P21397 , and P60880 , suggesting that future studies should explore potential moderators of these associations ( e.g. , ADHD subtype diagnoses , gender , exposure to environmental risk factors ) . We conclude with a discussion of these findings in relation to emerging themes relevant to future studies of the genetics of ADHD . A molecular basis underlying differences in the toxicity of botulinum serotypes A and E . Botulinum neurotoxins ( BoNTs ) block neurotransmitter release through their specific proteolysis of the proteins responsible for vesicle exocytosis . Paradoxically , two serotypes of BoNTs , A and E , cleave the same molecule , synaptosome-associated protein with relative molecular mass 25K ( P60880 ) , and yet they cause synaptic blockade with very different properties . Here we compared the action of BoNTs A and E on the plasma membrane fusion machinery composed of syntaxin and P60880 . We now show that the DB00083 -cleaved P60880 maintains its association with two syntaxin isoforms in vitro , which is mirrored by retention of P60880 on the plasma membrane in vivo . In contrast , BoNT/E severely compromises the ability of P60880 to bind the plasma membrane syntaxin isoforms , leading to dissociation of P60880 . The distinct properties of botulinum intoxication , therefore , can result from the ability of shortened P60880 to maintain its association with syntaxins-in the case of DB00083 poisoning resulting in unproductive syntaxin/ P60880 complexes that impede vesicle exocytosis .

MH_train_4

MH_train_4

MH_train_4

interacts_with DB06813?

multiple_choice

P21554 cannabinoid receptor deficiency promotes cardiac remodeling induced by pressure overload in mice . BACKGROUND : The endocannabinoid system is known to play a role in regulating myocardial contractility , but the influence of cannabinoid receptor 1 ( P21554 ) deficiency on chronic heart failure ( CHF ) remains unclear . In this study we attempted to investigate the effect of P21554 deficiency on CHF induced by pressure overload and the possible mechanisms involved . METHODS AND RESULTS : A CHF model was created by transverse aortic constriction ( TAC ) in both P21554 knockout mice and wild-type mice . P21554 knockout mice showed a marked increase of mortality due to CHF from 4 to 8 weeks after TAC ( p=0.021 ) . Five weeks after TAC , in contrast to wild-type mice , P21554 knockout mice had a higher left ventricular ( LV ) end-diastolic pressure , lower rate of LV pressure change ( ± dp/dt max ) , lower LV contractility index , and a larger heart weight to body weight ratio and lung weight to body weight ratio compared with wild-type mice ( all p < 0.05-0.001 ) . Phosphorylation of the epidermal growth factor receptor ( P00533 ) and mitogen-activated protein kinases ( O75791 and P29323 ) was higher in P21554 knockout mice than that in wild-type mice . In cultured neonatal rat cardiomyocytes , a P21554 agonist reduced DB02527 production stimulated by isoproterenol or forskolin , and suppressed phosphorylation of the P00533 , O75791 , and P29323 , while the inhibitory effect of a P21554 agonist on P00533 phosphorylation was abrogated by P21554 knockdown . CONCLUSION : These findings indicate that cannabinoid receptor 1 inactivation promotes cardiac remodeling by enhancing the activity of the epidermal growth factor receptor and mitogen-activated protein kinases . Single agent and combination studies of pralatrexate and molecular correlates of sensitivity . BACKGROUND : DB06813 is a dihydrofolate reductase ( P00374 ) inhibitor with high affinity for reduced folate carrier 1 ( P41440 -1 ) and folylpolyglutamate synthetase ( Q05932 ) , resulting in extensive internalization and accumulation in tumour cells . DB06813 is approved in the US for the treatment of relapsed or refractory peripheral T-cell lymphoma and is being investigated in various malignancies . Here , we evaluated molecular correlates of sensitivity to pralatrexate and explored combinations with a variety of anticancer agents . METHODS : Antiproliferative effects of pralatrexate were evaluated in 15 human-cancer cell lines using the MTT assay . Gene expression was evaluated using qRT-PCR . RESULTS : DB06813 and methotrexate had a similar pattern of cytotoxicity , pralatrexate being more potent . DB06813 potentiated the effects of platinum drugs , antimetabolites and P00533 inhibitors . Dose- and time-dependent cytotoxicity of pralatrexate correlated with high mRNA expression of Q05932 . Acquired resistance to pralatrexate was associated with decreased P41440 -1 expression , whereas methotrexate resistance correlated with increased P00374 expression , suggesting different mechanisms of acquired resistance . CONCLUSION : DB06813 was more potent than methotrexate in a panel of solid tumour lines . Our findings support the further clinical development of pralatrexate in combination with certain cytotoxics and targeted therapies , and suggest that P41440 -1 , Q05932 and P00374 may be potential biomarkers of outcome . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . On the relevance of glycolysis process on brain gliomas . The proposed analysis considers aspects of both statistical and biological validation of the glycolysis effect on brain gliomas , at both genomic and metabolic level . In particular , two independent datasets are analyzed in parallel , one engaging genomic ( Microarray Expression ) data and the other metabolomic ( Magnetic Resonance Spectroscopy Imaging ) data . The aim of this study is twofold . First to show that , apart from the already studied genes ( markers ) , other genes such as those involved in the human cell glycolysis significantly contribute in gliomas discrimination . Second , to demonstrate how the glycolysis process can open new ways towards the design of patient-specific therapeutic protocols . The results of our analysis demonstrate that the combination of genes participating in the glycolytic process ( P04075 , P09972 , P09104 , P04406 , P52789 , P00338 , P07195 , P40925 , P11177 , P08237 , P06744 , P00558 , P36871 and P30613 ) with the already known tumor suppressors ( P60484 , Rb , P04637 ) , oncogenes ( P11802 , P00533 , PDGF ) and Q9BYW2 , enhance the discrimination of low versus high-grade gliomas providing high prediction ability in a cross-validated framework . Following these results and supported by the biological effect of glycolytic genes on cancer cells , we address the study of glycolysis for the development of new treatment protocols . DB06813 : a novel synthetic antifolate for relapsed or refractory peripheral T-cell lymphoma and other potential uses . PURPOSE : The pharmacology , pharmacokinetics , clinical trials , adverse effects , dosage , and economic considerations of pralatrexate ( DB06813 ) are reviewed . SUMMARY : Peripheral T-cell lymphoma ( PTCL ) comprises approximately 15-20 % of all aggressive lymphomas and 5-10 % of all non-Hodgkin 's lymphomas . Advanced PTCL is often refractory to traditional first-line chemotherapy regimens . DB06813 was developed as a synthetic folate analog antimetabolite that competitively inhibits dihydrofolate reductase ( P00374 ) . This results in the depletion of thymidine , leading to interference with deoxyribonucleic acid synthesis and cancer cell death . DB06813 has a higher potency than methotrexate and edatrexate ( EDX ) . The efficacy and safety of DB06813 have been demonstrated in the PROPEL trial , a prospective phase II trial in patients with relapsed or refractory PTCL . Patients with prior stem cell transplantation receiving DB06813 also had similar response rates ( RRs ) . DB06813 was investigated on the treatment of relapsed or refractory cutaneous T-cell lymphoma , previously treated advanced non-small cell lung cancer and other solid malignancies . DB06813 has similar side effects to other P00374 inhibitors . The most common side effect of DB06813 is mucositis . The recommended dose of DB06813 is 30 mg/m(2) weekly once for 6 weeks in 7-week cycle until disease progresses or unacceptable toxicity for PTCL and may require dose reduction or discontinuation . Patients should be supplemented with oral folic acid and intramuscular vitamin B(12) injections . CONCLUSION : DB06813 provides clinical benefit to patients with relapsed or refractory PTCL with durable complete and partial responses in patients who had not responded to multiple prior treatment regimens . Sanguinarine suppresses basal-like breast cancer growth through dihydrofolate reductase inhibition . Basal-like breast cancer ( BLBC ) remains a great challenge because of its clinically aggressive nature and lack of effective targeted therapy . We analyzed the potential anti-neoplastic effects of sanguinarine , a natural benzophenanthridine alkaloid , against BLBC cells . Sanguinarine treatment resulted in a reduction of cell migration , in a dose-dependent inhibition of cell viability and in the induction of cell death by apoptosis in both human ( MDA-MB-231 cells ) and mouse ( A17 cells ) in vitro models of BLBC . In vivo experiments demonstrated that oral administration of sanguinarine reduced the development and growth of A17 transplantable tumors in FVB syngeneic mice . Western blotting analysis revealed that suppression of BLBC growth by sanguinarine was correlated with a concurrent upregulation of p27 and downregulation of cyclin D1 and with the inhibition of P40763 activation . In addition , we identified sanguinarine as a potent inhibitor of dihydrofolate reductase ( P00374 ) , able to impair enzyme activity even in methotrexate resistant MDA-MB-231 cells . These results provide evidence that sanguinarine is a promising anticancer drug for the treatment of BLBC . Inhibition of histone deacetylase activity is a novel function of the antifolate drug methotrexate . DB00563 ( MTX ) is a dihydrofolate reductase ( P00374 ) inhibitor widely used for treating human cancers , and overexpression of histone deacetylase ( HDAC ) is usually found in tumors . HDAC inhibitors ( HDACi ) can reactivate tumor suppressor genes and serve as potential anti-cancer drugs . In this study , we found that MTX shared structural similarity with some HDACi and molecular modeling showed that MTX indeed docks into the active site of HDLP , a bacterial homologue of HDAC . Subsequent in vitro assay demonstrated MTX 's inhibition on HDAC activity in human cancer cells . The global acetylation of histone H3 was also induced by MTX . Moreover , MTX inhibited immunoprecipitated Q13547 /2 activity but not their protein levels . This study provides evidence that MTX inhibits HDAC activity . Spline-fitting with a genetic algorithm : a method for developing classification structure-activity relationships . Classification methods allow for the development of structure-activity relationship models when the target property is categorical rather than continuous . We describe a classification method which fits descriptor splines to activities , with descriptors selected using a genetic algorithm . This method , which we identify as SFGA , is compared to the well-established techniques of recursive partitioning ( RP ) and soft independent modeling by class analogy ( SIMCA ) using five series of compounds : cyclooxygenase-2 ( P35354 ) inhibitors , benzodiazepine receptor ( BZR ) ligands , estrogen receptor ( ER ) ligands , dihydrofolate reductase ( P00374 ) inhibitors , and monoamine oxidase ( MAO ) inhibitors . Only 1-D and 2-D descriptors were used . Approximately 40 % of compounds in each series were assigned to a test set , " cherry-picked " from the complete set such that they lie outside the training set as much as possible . SFGA produced models that were more predictive for all but the P00374 set , for which SIMCA was most predictive . RP gave the least predictive models for all but the MAO set . A similar trend was observed when using training and test sets to which compounds were randomly assigned and when gradually eliminating compounds from the ( designed ) training set . The stability of models was examined for the random and reduced sets , where stability means that classification statistics and the selected descriptors are similar for models derived from different sets . Here , SIMCA produced the most stable models , followed by SFGA and RP . We show that a consensus approach that combines all three methods outperforms the single best model for all data sets . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Identification and analysis of specific chromosomal region adjacent to exogenous Dhfr-amplified region in Chinese hamster ovary cell genome . Chinese hamster ovary ( CHO ) cells are widely used for the stable production of recombinant proteins . Gene amplification techniques are frequently used to improve of protein production , and the dihydrofolate reductase ( P00374 ) gene amplification system is most widely used in the CHO cell line . We previously constructed a CHO genomic bacterial artificial chromosome ( BAC ) library from a mouse Dhfr-amplified CHO DR1000L-4N cell line and one BAC clone ( Cg0031N14 ) containing the CHO genomic DNA sequence adjacent to Dhfr was selected . To identify the specific chromosomal region adjacent to the exogenous Dhfr-amplified region in the CHO cell genome , we performed further screening of BAC clones to obtain other Dhfr-amplified regions in the CHO genome . From the screening by high-density replica filter hybridization using a digoxigenin-labeled pSV2-dhfr/hGM- P04141 probe , we obtained 8 new BAC clones containing a Dhfr-amplified region . To define the structures of the 8 BAC clones , Southern blot analysis , BAC end sequencing and fluorescence in situ hybridization ( Q5TCZ1 ) were performed . These results revealed that all the selected BAC clones contained a large palindrome structure with a small inverted repeat in the junction region . This suggests that the obtained amplicon structure in the Dhfr-amplified region in the CHO genome plays an important role in exogenous gene amplification . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 , CYP2B1/2 , P05181 , CYP3A , catechol-O-methyltransferase ( P21964 ) and microsomal glutathione transferase 1 ( P10620 ) . All but actin and soluble P21964 were positively detected at ≥1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 . The binding correlated well with inhibition of CYP activities , except for P05181 whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 antibodies showed a significant reduction in binding at ≥1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults . [ Genetic and clinical and pathological characteristics of breast cancer in premenopausal and postmenopausal women ] . This study involved 525 breast cancer ( BC ) patients of P24752 -4N0-2M0 stages at the age of 35 years and older . Significant differences in clinical and pathological characteristics between premenopausal and postmenopausal BC patients were found . Mostly marked differences were shown for positive lymph node correlation with distant metastasis , multicentric growth and local recurrence depending on menopause status . The prevalence of various morphological structures in primary tumors was appeared to be associated with different forms of tumor progression in pre- and postmenopausal women . We have studied polymorphisms in 15 genes involved in major cancer related pathways ( apoptosis , interleukins , folate metabolism enzymes genes ) . We found that variant genotypes of P42898 and P00374 genes were associated with an increased BC risk among premenopausal women while polymorphism in Q14116 , p53 genes were associated with BC among postmenopausal women . These results demonstrate novel biological information , which points the different mechanisms contributed to breast cancer progression in premenopausal and postmenopausal women . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . A phase II study of pralatrexate with vitamin B12 and folic acid supplementation for previously treated recurrent and/or metastatic head and neck squamous cell cancer . BACKGROUND : DB06813 ( Fotolyn(TM) ; Allos Therapeutics Inc. ) is an antifolate dihydrofolate reductase ( P00374 ) inhibitor . We conducted a phase II study of pralatrexate with folic acid and B12 supplementation in patients with recurrent and/or metastatic head and neck squamous cell cancer ( R/M HNSCC ) . PATIENTS AND METHODS : This was a single-arm , Simon optimal two stage phase II study . Patients with R/M HNSCC previously treated with chemotherapy were eligible . The study was initiated with a dosing schedule of pralatrexate 190 mg/m(2) biweekly on a 4-week cycle with vitamin supplementation . Due to toxicity concerns , the dosing was modified to 30 mg/m(2) weekly for 3 weeks in a 4-week cycle with vitamin supplementation . Radiologic imaging was to be obtained about every 2 cycles . RESULTS : Thirteen subjects were enrolled ; 12 were treated . Seven of the twelve patients had previously received ≥2 lines of chemotherapy . The most common grade 3 toxicity was mucositis ( 3 patients ) . Seven patients did not complete two cycles of therapy due to progression of disease ( 4 ) , toxicity ( 1 ) , death ( 1 ) , and withdrawal of consent ( 1 ) . Two deaths occurred : one due to disease progression and the other was an unwitnessed event that was possibly related to pralatrexate . No clinical activity was observed . The median overall survival was 3.1 months . The study was closed early due to lack of efficacy . CONCLUSIONS : DB06813 does not possess clinical activity against previously treated R/M HNSCC . Evaluation of pralatrexate in other clinical settings of HNSCC management with special considerations for drug toxicity may be warranted . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Rational design of an P01133 - Q14116 fusion protein : implication for developing tumor therapeutics . Q14116 ( Q14116 ) is a proinflammatory cytokine . This protein has a role in regulating immune responses and exhibits significant anti-tumor activities . Epidermal growth factor ( P01133 ) is an important growth factor that plays a central role in the regulation of cell cycle and differentiation . It was proposed that a targeted delivery of Q14116 by generation of Q14116 - P01133 fusion protein might decrease adverse effects and result in enhancing cytotoxic and antitumor activities . In the present study , a fusion protein , consisting of P00533 binding domain fused to human Q14116 mature peptide via a linker peptide of ( DB00145 (4) DB00133 ) 3 , was constructed and expressed in the insect cell line Sf9 using Bac-to-Bac baculovirus expression system . We showed that the purified recombinant fusion protein induced similar levels of P01579 to that of native Q14116 protein in human PBMC in the presence of ConA . Furthermore , P01133 receptor competitive test in human epithelial cancer A431 cell line showed that P01133 - Q14116 fusion protein can specifically bind with P00533 by competing with native P01133 protein . These suggest that this rationally designed protein can be further developed as novel tumor therapeutics . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . In vitro and in vivo biological activities of a novel nonpolyglutamable anti-folate , MX-68 . MX-68 is a newly synthesized anti-folate , chemically designed not to undergo intracellular polyglutamation and to have increased affinity to dihydrofolate reductase ( P00374 ) . In the present study , we examined the in vitro and in vivo biological activities of MX-68 compared with methotrexate ( MTX ) which forms several polyglutamates intracellularly . MX-68 dose-dependently inhibited the proliferation of PHA- , anti-CD3- , or PMA plus ionomycin-stimulated peripheral blood mononuclear cells ( PBMC ) and endothelial cells ( EC ) from normal subjects as well as P01584 - or P01375 alpha-stimulated synovial fibroblastic cells ( SC ) from rheumatoid arthritis ( RA ) patients . Coaddition of folinic acid completely reversed the anti-proliferative effects of both MX-68 and MTX . Although the anti-proliferative activities of MX-68 were almost comparable to those of MTX , the washout study clearly showed the characteristic nature of MX-68 . When drugs were removed during culture , the suppressive effect of MX-68 completely disappeared , whereas suppression by MTX was merely weakened . MX-68 dramatically suppressed the onset of collagen-induced arthritis ( CIA ) in mice when the drug was orally administered three times a week. starting from the day of first immunization . In this model , 2 mg/kg of MX-68 was sufficient to completely suppress arthritis , whereas suppression by the same dose of MTX was partial . These lines of evidence suggest that polyglutamation is not always a prerequisite in the anti-rheumatic effects of anti-folate . In addition , since intracellular accumulation of polyglutamates is thought to have adverse effects , MX-68 may become a more potent and less toxic anti-rheumatic drug than MTX . [ Cellular adhesion signal transduction network of tumor necrosis factor-alpha induced hepatocellular carcinoma cells ] . OBJECTIVE : To systemically explore the cellular adhesion signal transduction network of tumor necrosis factor-alpha ( P01375 -α ) -induced hepatocellular carcinoma cells with bioinformatics tools . METHODS : Published microarray dataset of P01375 -α-induced HepG2 , human transcription factor database HTRI and human protein-protein interaction database HPRD were used to construct and analyze the signal transduction network . RESULTS : In the signal transduction network , MYC and SP1 were the key nodes of signaling transduction . Several genes from the network were closely related with cellular adhesion. P00533 ( P00533 ) is a possible key gene of effectively regulating cellular adhesion during the induction of P01375 -α . CONCLUSION : P00533 is a possible key gene for P01375 -α-induced metastasis of hepatocellular carcinoma . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Insights into antifolate resistance from malarial P00374 -TS structures . Plasmodium falciparum dihydrofolate reductase-thymidylate synthase ( PfDHFR-TS ) is an important target of antimalarial drugs . The efficacy of this class of P00374 -inhibitor drugs is now compromised because of mutations that prevent drug binding yet retain enzyme activity . The crystal structures of PfDHFR-TS from the wild type ( TM4/8.2 ) and the quadruple drug-resistant mutant ( V1/S ) strains , in complex with a potent inhibitor WR99210 , as well as the resistant double mutant ( P04264 P21554 ) with the antimalarial pyrimethamine , reveal features for overcoming resistance . In contrast to pyrimethamine , the flexible side chain of WR99210 can adopt a conformation that fits well in the active site , thereby contributing to binding . The single-chain bifunctional PfDHFR-TS has a helical insert between the P00374 and TS domains that is involved in dimerization and domain organization . Moreover , positively charged grooves on the surface of the dimer suggest a function in channeling of substrate from TS to P00374 active sites . These features provide possible approaches for the design of new drugs to overcome antifolate resistance . P00374 protects endothelial nitric oxide synthase from uncoupling in tetrahydrobiopterin deficiency . DB00360 ( BH4 ) is a required cofactor for the synthesis of NO by endothelial nitric oxide synthase ( P29474 ) , and endothelial BH4 bioavailability is a critical factor in regulating the balance between NO and superoxide production ( P29474 coupling ) . Biosynthesis of BH4 is determined by the activity of GTP-cyclohydrolase I ( GTPCH ) . However , BH4 levels may also be influenced by oxidation , forming 7,8-dihydrobiopterin ( BH2 ) , which promotes P29474 uncoupling . Conversely , dihydrofolate reductase ( P00374 ) can regenerate BH4 from BH2 , but whether P00374 is functionally important in maintaining P29474 coupling remains unclear . To investigate the mechanism by which P00374 might regulate P29474 coupling in vivo , we treated wild-type , BH4-deficient ( hph-1 ) , and GTPCH-overexpressing ( P30793 -Tg ) mice with methotrexate ( MTX ) , to inhibit BH4 recycling by P00374 . MTX treatment resulted in a striking elevation in BH2 and a decreased BH4:BH2 ratio in the aortas of wild-type mice . These effects were magnified in hph-1 but diminished in P30793 -Tg mice . Attenuated P29474 activity was observed in MTX-treated hph-1 but not wild-type or P30793 -Tg mouse lung , suggesting that inhibition of P00374 in BH4-deficient states leads to P29474 uncoupling . Taken together , these data reveal a key role for P00374 in regulating the BH4 vs BH2 ratio and P29474 coupling under conditions of low total biopterin availability in vivo . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Significance of interleukin-6 signaling in the resistance of pharyngeal cancer to irradiation and the epidermal growth factor receptor inhibitor . PURPOSE : Tumor eradication by chemoradiotherapy for pharyngeal cancer has not been particularly successful . Targeting epithelial growth factor receptor ( P00533 ) could be a potential treatment strategy providing additional benefits , but only a subset of these tumors gives a clinically significant response to P00533 inhibitors . The aim has been to identify the role of interleukin-6 ( P05231 ) signaling and its predictive power in the treatment response of pharyngeal cancer . METHODS AND MATERIALS : Human pharyngeal cancer cell lines , including the hypopharyngeal cancer cell line FaDu and its derived cell line FaDu-C225-R , were selected . Changes in tumor growth , response to treatment , and responsible signaling pathway were investigated in vitro . Furthermore , 95 pharyngeal cancer tissue specimens were analyzed by immunohistochemical staining , and correlations were made between levels of P05231 , P05231 receptor ( IL-6R ) , p-AKT , and p- P40763 expression and the clinical outcome of patients . RESULTS : In vitro , either extrinsic P05231 stimulation of cancer cells or intrinsically activated P05231 signaling detected in FADu-C225-R cells results in resistance to irradiation and P00533 inhibitor . Blocking P05231 signaling attenuated aggressive tumor behavior and sensitized the cells to treatments . The responsible mechanisms included decreased p- P40763 , less nuclear translocation of P00533 , and subsequently attenuated epithelial-mesenchymal transition . Regarding clinical data , staining of p- P40763 and P05231 was significantly linked with lower response rates to treatments and shorter survival in pharyngeal cancer patients . CONCLUSIONS : P05231 and p- P40763 may be significant predictors of pharyngeal carcinoma , and regulating P05231 signaling can be considered a promising therapeutic approach . Evaluation of the pharmacokinetics , preclinical and clinical efficacy of pralatrexate for the treatment of T-cell lymphoma . INTRODUCTION : Peripheral T-cell lymphomas ( PTCLs ) are a heterogeneous group of T-cell neoplasms . Most patients with PTCL have a poor outcome with conventional therapies and are not cured without stem-cell transplantation . DB06813 , a novel antifolate chemotherapeutic agent , was rationally designed to impede folate metabolism by inhibiting dihydrofolate reductase ( P00374 ) and to be more efficiently internalized into tumor cells . DB06813 is the first drug that is FDA approved for patients with relapsed and refractory PTCL . AREAS COVERED : DB06813 has been used as a single agent and in combination with other agents in clinical trials for non-Hodgkin 's lymphoma and Hodgkin 's disease as well as in solid tumors . This review will cover the development of pralatrexate , the pharmacokinetics of pralatrexate , preclinical findings with pralatrexate and clinical studies of pralatrexate in hematologic malignancies . EXPERT OPINION : DB06813 has significant activity in vitro , and in early Phase I/II trials , responses were noted in patients with aggressive T-cell lymphomas . The DB06813 in Patients with Relapsed or Refractory Peripheral T-Cell Lymphoma trial demonstrated the activity of pralatrexate across a spectrum of heavily pretreated patients with different aggressive T-cell lymphoma subtypes , and studies in cutaneous T-cell lymphoma have shown efficacy at different doses and schedules . The most frequent adverse events in these trials were mucositis , reversible thrombocytopenia and fatigue . Class I histone deacetylase activity is required for proliferation of renal epithelial cells . The process of renal regeneration after acute kidney injury is thought to recapitulate renal development , and proliferation of renal proximal tubular cells ( RPTCs ) is a critical step in the regenerative response . Recent studies indicate that class I histone deacetylases ( HDACs ) are required for embryonic kidney gene expression , growth , and differentiation . The role and underlying mechanisms of class I HDAC activation in RPTC proliferation , however , remain unclear . In this study , we used cultured RPTCs to examine this issue since four class I HDAC isoforms ( 1 , 2 , 3 , and 8 ) are abundantly expressed in this cell type . Blocking class I HDAC activity with a highly selective inhibitor , MS-275 , induced global histone H3 hyperacetylation , reduced RPTC proliferation , and diminished expression of cyclin D1 and proliferating cell nuclear antigen . Silencing Q13547 , 3 , or 8 with small interfering RNA resulted in similar biological effects . Activation of epidermal growth factor receptor ( P00533 ) and signal transducers and activators of transcription 3 ( P40763 ) was required for RPTC proliferation , and P40763 functioned downstream of P00533 . Treatment with MS-275 or knockdown of Q13547 , 3 , or 8 suppressed P00533 expression and phosphorylation , and silencing Q13547 and 3 also reduced P40763 phosphorylation . However , Q92769 downregulation did not affect RPTC proliferation and phosphorylation of P00533 and P40763 . Collectively , these data reveal a critical role of class I HDACs in mediating proliferation of renal epithelial cells through activation of the P00533 / P40763 signaling pathway . P10275 controls P00533 and P04626 gene expression at different levels in prostate cancer cell lines . P00533 or P04626 contributes to prostate cancer ( PCa ) progression by activating the androgen receptor ( AR ) in hormone-poor conditions . Here , we investigated the mechanisms by which androgens regulate P00533 and P04626 expression in PCa cells . In steroid-depleted medium ( SDM ) , P00533 protein was less abundant in androgen-sensitive LNCaP than in androgen ablation-resistant 22Rv1 cells , whereas transcript levels were similar . DB02901 ( DB02901 ) treatment increased both P00533 mRNA and protein levels and stimulated RNA polymerase II recruitment to the P00533 gene promoter , whereas it decreased P04626 transcript and protein levels in LNCaP cells . DB02901 altered neither P00533 or P04626 levels nor the abundance of prostate-specific antigen ( PSA ) , TMEPA1 , or O15393 mRNAs in 22Rv1 cells , which express the full-length and a shorter AR isoform deleted from the COOH-terminal domain ( ARDeltaCTD ) . The contribution of both AR isoforms to the expression of these genes was assessed by small interfering RNAs targeting only the full-length or both AR isoforms . Silencing of both isoforms strongly reduced PSA , TMEPA1 , and O15393 transcript levels . Inhibition of both AR isoforms did not affect P00533 and P04626 transcript levels but decreased P00533 and increased P04626 protein levels . Proliferation of 22Rv1 cells in SDM was inhibited in the absence of AR and ARDeltaCTD . A further decrease was obtained with PKI166 , an P00533 / P04626 kinase inhibitor . Overall , we showed that ARDeltaCTD is responsible for constitutive P00533 expression and P04626 repression in 22Rv1 cells and that ARDeltaCTD and tyrosine kinase receptors are necessary for sustained 22Rv1 cell growth . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Distinct mechanistic activity profile of pralatrexate in comparison to other antifolates in in vitro and in vivo models of human cancers . PURPOSE : This study evaluated mechanistic differences of pralatrexate , methotrexate , and pemetrexed . METHODS : Inhibition of dihydrofolate reductase ( P00374 ) was quantified using recombinant human P00374 . Cellular uptake and folylpolyglutamate synthetase ( Q05932 ) activity were determined using radiolabeled pralatrexate , methotrexate , and pemetrexed in NCI-H460 non-small cell lung cancer ( NSCLC ) cells . The tumor growth inhibition ( TGI ) was assessed using MV522 and NCI-H460 human NSCLC xenografts . RESULTS : Apparent K ( i ) values for P00374 inhibition were 45 , 26 , and > 200 nM for pralatrexate , methotrexate , and pemetrexed , respectively . A significantly greater percentage of radiolabeled pralatrexate entered the cells and was polyglutamylatated relative to methotrexate or pemetrexed . In vivo , pralatrexate showed superior anti-tumor activity in both NSCLC models , with more effective dose-dependent TGI in the more rapidly growing NCI-H460 xenografts . CONCLUSIONS : DB06813 demonstrated a distinct mechanistic and anti-tumor activity profile relative to methotrexate and pemetrexed . DB06813 exhibited enhanced cellular uptake and increased polyglutamylation , which correlated with increased TGI in NSCLC xenograft models .

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Role of monoamine oxidases in the exaggerated 5-hydroxytryptamine-induced tension development of human isolated preeclamptic umbilical artery . We investigated the role(s) of monoamine oxidases ( MAOs ) on the altered 5-hydroxytryptamine ( 5-HT , serotonin ) -induced tension development of the isolated umbilical artery of preeclamptic pregnancy of Chinese women . An enhanced 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy was observed when compared with that of normal pregnancy . The enhanced component of 5-HT-induced tension development was eradicated by clorgyline ( a P21397 inhibitor ) . Blockade of P29474 ( endothelial isoform nitric oxide synthase ) ( N(omega)-nitro-L-arginine methyl ester ) , 5-HT transporter ( citalopram ) , 5-HT receptor subtypes ( 5HT2B , SB 204741 ; P28335 , RS 102221 ; P34969 , SB 269970 ) , and endothelium denudation of the umbilical artery of normal pregnancy mimicked the enhanced 5-HT-induced tension development as observed in the preeclamptic tissues . In contrast , no apparent changes in 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy were observed with the same pharmacological manipulations . A decreased protein expression levels of P21397 and P29474 ( no P35228 and P27338 expression was detected ) and no change in caveolin-1 and 5-HT transporter expression were demonstrated in the umbilical artery ( endothelium intact ) lysate of preeclamptic pregnancy , compared to that of the umbilical artery of normal pregnancy . Thus , in the umbilical artery of preeclamptic pregnancy , a decrease of P21397 and P29474 protein expression levels are probably associated with , or responsible for , the exaggerated 5-HT-induced tension development . DB06288 is a potent P34969 antagonist : relevance for antidepressant actions in vivo . RATIONALE : DB06288 is approved for clinical use in treating schizophrenia in a number of European countries and also for treating dysthymia , a mild form of depression , in Italy . DB06288 has also been demonstrated to be an antidepressant for patients with major depression in many clinical trials . In part because of the selective D(2)/D(3) receptor antagonist properties of amisulpride , it has long been widely assumed that dopaminergic modulation is the proximal event responsible for mediating its antidepressant and antipsychotic properties . OBJECTIVES : The purpose of these studies was to determine if amisulpride 's antidepressant actions are mediated by off-target interactions with other receptors . MATERIALS AND METHODS : We performed experiments that : ( 1 ) examined the pharmacological profile of amisulpride at a large number of central nervous system ( CNS ) molecular targets and , ( 2 ) after finding high potency antagonist affinity for human 5-HT(7a) serotonin receptors , characterized the actions of amisulpride as an antidepressant in wild-type and 5-HT(7) receptor knockout mice . RESULTS : We discovered that amisulpride was a potent competitive antagonist at 5-HT(7a) receptors and that interactions with no other molecular target investigated in this paper could explain its antidepressant actions in vivo . Significantly , and in contrast to their wild-type littermates , 5-HT(7) receptor knockout mice did not respond to amisulpride in two widely used rodent models of depression , the tail suspension test and the forced swim test . CONCLUSIONS : These results indicate that 5-HT(7a) receptor antagonism , and not D(2)/D(3) receptor antagonism , likely underlies the antidepressant actions of amisulpride . Serotonin P34969 receptors coupled to induction of interleukin-6 in human microglial MC-3 cells . Brain serotonin 5-HT(7) receptors are known to be expressed in neurons and astrocytes . We now report the presence of these receptors in a third type of cell , microglial cells . 5-Hydroxytryptamine ( 5-HT ) , 5-carboxamidotryptamine ( 5-CT ) , 5-methoxytryptamine ( 5-MeOT ) and 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) induced concentration-dependent stimulations of DB02527 accumulation in the human microglial MC-3 cell line . The maximal effect of 5-HT was 3.4+/-0.3-fold stimulation ( mean+/-S.E.M. , n=5 ) above basal levels . The rank order of agonist potency ( pEC50 values ) was 5-CT ( 7.09 ) > 5-HT ( 6.13 ) > or=5-MeOT ( 5.78 ) >> 8-OH-DPAT ( ca. 5 ) . The effect of 5-CT was inhibited in a concentration-dependent manner by the selective P34969 receptor antagonist SB-269970 ( pA2 value 9.03 ) . Western blot analysis revealed the presence of immunoreactive bands corresponding to the human P34969 receptor in extracts of MC-3 cells . The presence of two splice variants of the P34969 receptor ( P34969 (a/b) ) was visualized by reverse transcriptase-polymerase chain reaction ( RT-PCR ) analysis with specific primers . In real-time PCR studies , the mRNA for interleukin-6 ( P05231 ) was found to be increased by 2.5-fold in MC-3 cells after 1 h incubation with 5-CT ( 1 microM ) and this effect was fully blocked by the P34969 receptor antagonist SB-269970 ( 1 microM ) . These data show that functional P34969 receptors are present in human microglial MC-3 cells , suggesting that they are involved in neuroinflammatory processes . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Direct analysis of candidate genes in impulsive behaviours . Antisocial behaviour is both heterogeneous and the product of interacting genetic and environmental factors acting at different levels of causation . Heritability studies show that individual differences in predisposition to antisocial behaviour are transmitted vertically in families by genetic mechanisms . Owing to aetiological heterogeneity and complexity , study of a variety of other behavioural phenotypes may shed more light on the antecedents of antisocial behaviour than direct studies on antisocial behaviour . Identification of genetic vulnerability factors would clarify mechanisms of vulnerability and the role of the environment . Direct gene analysis and genetic linkage analysis have identified structural variants in genes involved in neurotransmitter function , and some progress has been made towards relating these genetic variants to antisocial personality and other behaviours . Thyroid hormone receptor variants can cause attention deficit/hyperactivity disorder , and a monoamine oxidase A variant leads to aggressive behaviour in one family . Direct gene analyses have revealed non-conservative amino acid substitutions and structural variants ( generally rare ) at P14416 , P35462 and P21917 dopamine receptors and P08908 , 5- Q13049 , P28335 and P34969 serotonin receptors . The stage is set to identify the phenotypic significance of these as well as genetic variants at other loci which may be relevant as candidate genes for antisocial behaviour and related behavioural differences . DB01296 improves cardiac function following trauma-hemorrhage by increased protein O-GlcNAcylation and attenuation of NF-{kappa}B signaling . We have previously demonstrated that in a rat model of trauma-hemorrhage ( T-H ) , glucosamine administration during resuscitation improved cardiac function , reduced circulating levels of inflammatory cytokines , and increased tissue levels of O-linked N-acetylglucosamine ( O-GlcNAc ) on proteins . The mechanism(s) by which glucosamine mediated its protective effect were not determined ; therefore , the goal of this study was to test the hypothesis that glucosamine treatment attenuated the activation of the nuclear factor-kappaB ( NF-kappaB ) signaling pathway in the heart via an increase in protein O-GlcNAc levels . Fasted male rats were subjected to T-H by bleeding to a mean arterial blood pressure of 40 mmHg for 90 min followed by resuscitation . DB01296 treatment during resuscitation significantly attenuated the T-H-induced increase in cardiac levels of P01375 and P05231 mRNA , IkappaB-alpha phosphorylation , NF-kappaB , NF-kappaB DNA binding activity , P05362 , and P05164 activity . LPS ( 2 microg/ml ) increased the levels of IkappaB-alpha phosphorylation , P01375 , P05362 , and NF-kappaB in primary cultured cardiomyocytes , which was significantly attenuated by glucosamine treatment and overexpression of O-GlcNAc transferase ; both interventions also significantly increased O-GlcNAc levels . In contrast , the transfection of neonatal rat ventricular myocytes with O15294 small-interfering RNA decreased O-GlcNAc transferase and O-GlcNAc levels and enhanced the LPS-induced increase in IkappaB-alpha phosphorylation . DB01296 treatment of macrophage cell line RAW 264.7 also increased O-GlcNAc levels and attenuated the LPS-induced activation of NF-kappaB . These results demonstrate that the modulation of O-GlcNAc levels alters the response of cardiomyocytes to the activation of the NF-kappaB pathway , which may contribute to the glucosamine-mediated improvement in cardiac function following hemorrhagic shock . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . Cytokine signaling via various transcription factors regulates receptor activator of nuclear factor ( NF ) -kappaB ligand ( O14788 ) -mediated osteoclast differentiation from monocyte/macrophage lineage cells involved in propagation and resolution of inflammatory bone destruction . Protein inhibitor of activated P40763 ( Q9Y6X2 ) was initially identified as a molecule that inhibits DNA binding of P40763 and regulates many transcription factors through distinct mechanisms . To analyze Q9Y6X2 function in osteoclasts in vivo , we have generated transgenic mice in which Q9Y6X2 is specifically expressed in the osteoclast lineage using the tartrate-resistant acid phosphatase ( TRAP ) gene promoter . Q9Y6X2 transgenic mice showed an osteopetrotic phenotype due to impairment of osteoclast differentiation . Overexpression of Q9Y6X2 in RAW264.7 cells suppressed O14788 -induced osteoclastogenesis by inhibiting the expression of c-Fos and O95644 . Interestingly , Q9Y6X2 inhibits the transcriptional activity of microphthalmia-associated transcription factor ( O75030 ) independent of sumoylation . Down-regulation of Q9Y6X2 markedly enhances O14788 -mediated osteoclastogenesis in RAW264.7 cells . Furthermore , overexpression of Q9Y6X2 in mouse primary osteoblast ( DB00925 ) , down-regulates O14788 expression induced by interleukin-6 ( P05231 ) cytokine family , and inhibits osteoclast formation from bone marrow macrophages ( BMMs ) in vitro coculture system . Down-regulation of Q9Y6X2 leads to the accelerated expression of O14788 in DB00925 stimulated with P05231 and soluble P05231 receptor ( sIL-6R ) . Taken together , our results clearly indicate that Q9Y6X2 negatively regulates O14788 -mediated osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblasts . DB06288 promotes cognitive flexibility in rats : the role of P34969 receptors . The antagonism of P34969 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug , amisulpride . It has been previously demonstrated that the selective P34969 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task ( ASST ) . Therefore , the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility . The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound 's action at P34969 receptors . Rats repeatedly exposed ( 1 h daily for 7 days ) to restraint stress demonstrated impaired performance on the extra-dimensional ( ED ) set-shifting stage of the ASST . DB06288 ( 3 mg/kg ) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group . The P34969 receptor agonist , AS19 ( 10 mg/kg ) , abolished the pro-cognitive efficacy of amisulpride ( 3 mg/kg ) . The present study suggests that the antagonism of P34969 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride . These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders . Regulation of P14061 and P18405 in lymphocytes . We previously reported lymphocyte expression of genes encoding enzymes required for steroid metabolism ; however , only 17beta-HSD and 5alpha-reductase showed significant enzyme activity . We now investigate regulation of lymphocyte expression for genes encoding 17beta-HSD and 5alpha-reductase . Cultured human T and B lymphoid cell lines and peripheral blood mononuclear cells were treated with known regulators of steroidogenic gene expression including forskolin , PMA , ionomycin , various steroids , interleukin ( IL ) -4 , and P05231 . Treatment with 10 or 50 microM forskolin resulted in a 20-60 % reduction of expression for P14061 ( encoding 17beta-HSD I ) in T and B lymphoid cell lines and peripheral blood mononuclear cells , although such a change was not observed in the expression of P18405 ( encoding 5alpha-reductase I ) . No significant changes were found when cells were treated for 24 h with various concentrations of PMA or ionomycin . Incubation with 10(-9) to 10(-7) M androstenedione or estradiol increased expression of P14061 , while testosterone decreased the expression of this gene . P18405 expression was increased in the presence of 5alpha- DB02901 although no consistent changes were observed when the cells were treated with testosterone . Other steroids , including dexamethasone , progesterone , and 6-hydroxypregnanolone , produced no effects on expression of either P14061 or P18405 . Treatment with 0.1-10 ng/ml of P05112 or P05231 also did not effect significant changes in gene expression . These data implicate the involvement of the DB02527 -protein kinase signal transduction pathway in regulating lymphocyte expression of P14061 . Furthermore , it appears that lymphocyte P14061 and P18405 are regulated to some extent by specific steroids . Serotonin skews human macrophage polarization through P41595 and P34969 . Besides its role as a neurotransmitter , serotonin ( 5-hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT(1-7) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 production , upregulated the expression of M2 polarization-associated genes ( P05120 , P07996 , Q9NY15 , Q86Y22 ) , and reduced the expression of M1-associated genes ( P08476 , P41597 , P39900 , P05121 , P29016 , O94788 ) . Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2( P09603 ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7) , whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies . S-sulfhydration of Q02750 leads to P09874 activation and DNA damage repair . The repair of DNA damage is fundamental to normal cell development and replication . Hydrogen sulfide ( H2S ) is a novel gasotransmitter that has been reported to protect cellular aging . Here , we show that H2S attenuates DNA damage in human endothelial cells and fibroblasts by S-sulfhydrating Q02750 at cysteine 341 , which leads to P09874 activation . H2S-induced Q02750 S-sulfhydration facilitates the translocation of phosphorylated P27361 /2 into nucleus , where it activates P09874 through direct interaction . Mutation of Q02750 cysteine 341 inhibits P29323 phosphorylation and P09874 activation . In the presence of H2S , activated P09874 recruits P18887 and P49916 to DNA breaks to mediate DNA damage repair , and cells are protected from senescence .

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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 . P19957 -kinase and Q96HU1 -kinase signaling cascades in Q9Y6W8 / Q9Y6W8 - and P10747 -costimulated T-cells have distinct functions between cell proliferation and P22301 production . Both Q9Y6W8 / Q9Y6W8 and P10747 provide positive costimulatory signals for T-cell activation , resulting in proliferation and cytokine production . In this study , we attempted to clarify the key signaling molecules in T-cell proliferation , and also P60568 and P22301 production , during T-cell activation by CD3 induced by costimulation with either Q9Y6W8 / Q9Y6W8 or P10747 . We examined the role of both the P19957 -kinase/Akt pathway and Q96HU1 kinase family members such as P27361 /2 , JNK , and p38 kinase in this process . P19957 -kinase and Erk1/2 were shown to potentially regulate primary T-cell activation and subsequent proliferation via both Q9Y6W8 / Q9Y6W8 - or P10747 -mediated costimulation and the Erk signaling cascade was essential for this proliferation induction and also for P60568 production . The JAK inhibitor , AG490 , inhibited this induction . Our studies indicate that P60568 is necessary for induction of T-cell proliferation and that the quantities of P60568 produced by Q9Y6W8 / Q9Y6W8 ligation are also sufficient for T-cells to proliferate . In contrast , inhibition of Akt and p38 , that are phosphorylated by both Q9Y6W8 / Q9Y6W8 and P10747 -ligation , could downregulate P22301 production but not T-cell proliferation . These data raise the interesting possibility that the signaling cascades between T-cell proliferation and P22301 production are regulated by different molecules in Q9Y6W8 / Q9Y6W8 - and P10747 -costimulated T-cells . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . Phosphodiesterase type 5 inhibitors for the treatment of erectile dysfunction in patients with diabetes mellitus . DB00203 , a phosphodiesterase 5 ( O76074 ) inhibitor , has become a first-line therapy for diabetic patients with erectile dysfunction ( ED ) . The efficacy in this subgroup , based on the Global Efficacy Question , is 56 % vs 84 % in a selected group of non-diabetic men with ED . Two novel O76074 inhibitors , tadalafil ( Lilly Q9Y6W8 ) and vardenafil ( Bayer ) , have recently completed efficacy and safety clinical trials in ' general ' and diabetic study populations and are now candidates for US FDA approval . A summary analysis of the phase three clinical trials of sildenafil , tadalafil and vardenafil in both study populations is presented to provide a foundation on which the evaluation of the role of the individual O76074 inhibitors for the treatment of patients with ED and DM can be built . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . High biochemical selectivity of tadalafil , sildenafil and vardenafil for human phosphodiesterase 5A1 ( O76074 ) over PDE11A4 suggests the absence of PDE11A4 cross-reaction in patients . The physiological role of phosphodiesterase (PDE)11 is unknown and its biochemical characteristics are poorly understood . We have expressed human DB00117 -tagged PDE11A4 and purified the enzyme to apparent homogeneity . PDE11A4 displays K(m) values of 0.97 microM for cGMP and 2.4 microM for DB02527 , and maximal velocities were 4- to 10-fold higher for DB02527 than for cGMP . Given the homology between PDE11 and O76074 , we have compared the biochemical potencies of tadalafil ( DB00820 , Lilly- Q9Y6W8 ) , vardenafil ( DB00862 , Bayer-GSK ) , and sildenafil ( Viagra , Pfizer Inc. ) for PDE11A4 and PDE5A1 . PDE5A1/PDE11A4 selectivities are 40- , 9300- , and 1000-fold for tadalafil , vardenafil , and sildenafil , respectively . This suggests that none of these three compounds is likely to crossreact with PDE11A4 in patients . T-cell regulation by casitas B-lineage lymphoma ( Cblb ) is a critical failsafe against autoimmune disease due to autoimmune regulator ( Aire ) deficiency . Autoimmune polyendocrinopathy syndrome type 1 ( Q96G61 ) results from homozygous Aire mutations that cripple thymic deletion of organ-specific T cells . The clinical course in man and mouse is characterized by high variability both in the latent period before onset of autoimmune disease and in the specific organs affected , but the reasons for this are unknown . Here we test the hypothesis that the latent period reflects the failsafe action of discrete postthymic mechanisms for imposing self-tolerance in peripheral T cells . Aire-deficient mice were crossed with mice of a uniform major histocompatibility complex ( MHC ) haplotype and genetic background carrying specific genetic defects in one of four distinct peripheral tolerance mechanisms : activation-induced cell death ( Fasl(gld/gld) ) , anergy and requirement for P10747 costimulation ( Cblb(-/-) ) , inhibition of Q9Y6W8 and T(FH) cells ( Rc3h1(san/san) ) , or decreased numbers of Foxp3(+) T regulatory cells ( Card11(unm/unm) ) . Cblb-deficiency was unique among these four in precipitating rapid clinical autoimmune disease when combined with Aire-deficiency , resulting in autoimmune exocrine pancreatitis with median age of survival of only 25 d . Massive lymphocytic infiltration selectively destroyed most of the exocrine acinar cells of the pancreas and submandibular salivary gland , and P01730 (+) and CD8(+) subsets were necessary and sufficient to transfer the disease . Intrinsic regulation of peripheral T cells by P22681 -B thus serves a uniquely critical role as a failsafe against clinical onset of autoimmune disease in O43918 deficiency , and multiple peripheral tolerance mechanisms may need to fail before onset of clinical autoimmunity to many organs . Genotoxicity and induction of DNA damage responsive genes by food-borne heterocyclic aromatic amines in human hepatoma HepG2 cells . Heterocyclic aromatic amines ( HAAs ) are potential human carcinogens formed in well-done meats and fish . The most abundant are 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline ( MeIQx ) , 2-Amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinoxaline ( 4,8-DiMeIQx ) and 2-Amino-3-methyl-3H-imidazo[4,5-f]quinoline ( IQ ) . HAAs exert genotoxic activity after metabolic transformation by CYP1A enzymes , that is well characterized , however the genomic and intervening responses are not well explored . We have examined cellular and genomic responses of human hepatoma HepG2 cells after 24h exposure to HAAs . Comet assay revealed increase in formation of DNA strand breaks by PhIP , MeIQx and IQ but not 4,8-DiMeIQx , whereas increased formation of micronuclei was not observed . The four HAAs up-regulated expression of genes encoding metabolic enzymes P04798 , P05177 and P22309 and expression of P04637 and its downstream regulated genes P38936 , GADD45α and Q07812 . Consistent with the up-regulation of P38936 and GADD45α the cell-cycle analysis showed arrest in S-phase by PhIP and IQ , and in P55008 -phase by 4,8-DiMeIQx and MeIQx . The results indicate that upon exposure to HAAs the cells respond with the cell-cycle arrest , which enables cells to repair the damage or eliminate them by apoptosis . However , elevated expression of P10415 and down-regulation of Q07812 may indicate that HAAs could suppress apoptosis meaning higher probability of damaged cells to survive and mutate . O75144 costimulation is required for T-cell encephalitogenicity . The interaction of Q9Y6W8 with its ligand on P25054 provides a costimulatory signal to previously activated T-cells . In these studies , we blocked the Q9Y6W8 : O75144 interaction with Q9Y6W8 -Ig during the in vitro activation of MBP-reactive transgenic P01730 (+) T-cells . The presence of Q9Y6W8 -Ig in these cultures inhibited the ability of the transgenic T-cells to transfer EAE , although they entered the brains of the recipient mice . Q9Y6W8 -Ig increased apoptosis in the transgenic T-cells , especially in the memory population . This enhanced apoptosis was accompanied by an increase in the Q07812 /BCL-2 mRNA ratio . Q9Y6W8 -Ig did not prevent P60568 production , demonstrating that P60568 production is O75144 independent . P01579 and P22301 production by the transgenic T-cells , however , was suppressed . Finally , Q9Y6W8 -Ig injection into mice after the first signs of EAE ameliorated clinical disease . Therefore , O75144 provides a signal distinct from P10747 costimulation that is required for the activation and viability of encephalitogenic T-cells . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . S-nitrosoglutathione modulates P61073 and Q9Y6W8 expression . The expression of P61073 , a membrane protein which is involved in the entry of HIV-1 , is down-modulated from the cell surface by Phorbol 12-myristate 13-acetate ( PMA ) and the Ca+ ionophore , Ionomycin . Inducible co-stimulator ( Q9Y6W8 ) , which contributes to lymphocyte proliferation , is up-regulated by PMA/Ionomycin . We examined the influence of S-nitrosoglutathione ( SNG ) , an inhibitor of Vacuolar H+-ATPase ( V-ATPase ) , on the expression of P61073 and Q9Y6W8 in PMA/Ionomycin-treated peripheral mononuclear cells ( PBMC ) , and of P61073 alone in lymphoid cell lines . In this report , we show that SNG interferes with both effects of PMA/Ionomycin , namely P61073 down-regulation and Q9Y6W8 up-regulation . These studies imply opposing roles of V-ATPase in the regulation of P61073 and Q9Y6W8 . The influence of SNG in modulating the susceptibility of T cells to HIV-1 and on their immune responses needs further investigation . 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 . Emerging oral drugs for erectile dysfunction . Erectile dysfunction ( ED ) is a common medical condition that affects the sexual life of millions of men worldwide . Many drugs are now available for the treatment of ED , with oral pharmacotherapy representing the first-line option for most patients . DB00203 citrate , an inhibitor of the enzyme phosphodiesterase type 5 ( O76074 ) , is the most widely prescribed oral agent and has a very satisfactory efficacy-safety profile in all patient categories . DB00820 ( DB00820 ; Eli Lilly & Co. , Q9Y6W8 ) and vardenafil ( DB00862 ; Bayer Pharmaceuticals , GlaxoSmithKline ) are new O76074 inhibitors that have recently been approved worldwide . Both have been associated with significant positive efficacy-safety profiles . DB00714 sublingual is a dopamine D1 and D2 receptor agonist , which has been approved for marketing in Europe . It is best selected for treating patients with mild-to-moderate ED , but it is seldom used in clinical practice due to its limited efficacy and side effects , particularly nausea . Patients who do not respond to oral pharmacotherapy or who are unable to use it are appropriate candidates for intracavernosal and intraurethral therapy . The efficacy of second-line treatment is high , but the attrition rate remains significant . For the purpose of this review , clinical and pharmacological analysis focuses on the recent advances in the field of oral therapy , including O76074 inhibitors and sublingual apomorphine . Induction , binding specificity and function of human Q9Y6W8 . Recently , we have identified the inducible co-stimulator ( Q9Y6W8 ) , an activation-dependent , T cell-specific cell surface molecule related to P10747 and P16410 . Detailed analysis of human Q9Y6W8 presented here shows that it is a 55-60-kDa homodimer with differently N-glycosylated subunits of 27 and 29 kDa . Q9Y6W8 requires both phorbol 12-myristate 13-acetate and ionomycin for full induction , and is sensitive to DB00091 . Q9Y6W8 is up-regulated early on all T cells , including the P10747 - subset , and continues to be expressed into later phases of T cell activation . On stimulation of T cells by antigen-presenting cells , the P10747 / P33681 , but not the P29965 / P25942 pathway is critically involved in the induction of Q9Y6W8 . Q9Y6W8 does not bind to P33681 -1 or P33681 -2 , and P10747 does not bind to O75144 ; thus the P10747 and Q9Y6W8 pathways do not cross-interact on the cell surface . In vivo , Q9Y6W8 is expressed in the medulla of the fetal and newborn thymus , in the T cell zones of tonsils and lymph nodes , and in the apical light zones of germinal centers ( predominant expression ) . Functionally , Q9Y6W8 co-induces a variety of cytokines including P05112 , P05113 , P05231 , P01579 , P01375 , GM- P04141 , but not P60568 , and superinduces P22301 . Furthermore , Q9Y6W8 co-stimulation prevents the apoptosis of pre-activated T cells . The human Q9Y6W8 gene maps to chromosome 2q33 - 34 . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Pharmacokinetics of [(18)F]fluoroalkyl derivatives of dihydrotetrabenazine in rat and monkey brain . The specific binding and regional brain pharmacokinetics of new fluorine-18 ( [ (18)F ] ) -labeled radioligands for the vesicular monoamine transporter ( Q05940 ) were examined in the rat and primate brain . In the rat , 9-[(18)F]fluoropropyl-(+/-)-9-O-desmethyldihydrotetrabenazine ( [(18)F]FP-(+/-)-DTBZ ) showed better specific binding in the striatum than either (+)-[(11)C]dihydrotetrabenazine ( (+)-[(11)C]DTBZ ) or 9-[(18)F]fluoroethyl-(+/-)-9-O-desmethyldihydrotetrabenazine ( [(18)F]FE-(+/-)-DTBZ ) . Using microPET , the regional brain pharmacokinetics of [(18)F]FE-(+/-)-DTBZ , [(18)F]FP-(+/-)-DTBZ and (+)-[(11)C]DTBZ were examined in the same monkey brain . (+)-[(11)C]DTBZ and [(18)F]FP-(+/-)-DTBZ showed similar brain uptakes and pharmacokinetics , with similar maximum striatum-to-cerebellum ratios ( STR/ P22681 =5.24 and 5.15 , respectively ) that were significantly better than obtained for [(18)F]FE-(+/-)-DTBZ ( STR/ P22681 =2.55 ) . Striatal distribution volume ratios calculated using Logan plot analysis confirmed the better specific binding for the fluoropropyl compound [ distribution volume ratio (DVR)=3.32 ] vs. the fluoroethyl compound ( DVR=2.37 ) . Using the resolved single active isomer of the fluoropropyl compound , [(18)F]FP-(+)-DTBZ , even better specific to nonspecific distribution was obtained , yielding the highest distribution volume ratio ( DVR=6.2 ) yet obtained for a Q05940 ligand in any species . The binding of [(18)F]FP-(+)-DTBZ to the Q05940 was shown to be reversible by administration of a competing dose of unlabeled tetrabenazine . Metabolic defluorination was slow and minor for the [(18)F]fluoroalkyl-DTBZ ligands . The characteristics of high specific binding ratio , reversibility , metabolic stability and longer half-life of the radionuclide make [(18)F]FP-(+)-DTBZ a promising alternative Q05940 radioligand suitable for widespread use in human positron emission tomography studies of monoaminergic innervation of the brain . Increased frequencies of nuocytes in peripheral blood from patients with Graves ' hyperthyroidism . Newly identified nuocytes play an important role in Th2 cell mediated immunity such as protective immune responses to helminth parasites , allergic asthma and chronic rhinosinusitis . However , the contributions of nuocytes in the occurrence and development of Graves ' hyperthyroidism remains unknown . Previous studies found that there was a predominant Th2 phenotype in patients with Graves ' hyperthyroidism , it might relate to polarization of nuocytes . Nuocytes were defined by transcription factor RORα , various cell surface markers ( T1/ST2 , Q9NRM6 , Q9Y6W8 , P08575 ) and associated cytokines . In this study , these cells related genes or molecules in PBMC from patients with Graves ' hyperthyroidism were measured , and the potential correlation between them was analyzed . The expression levels of T1/ST2 , Q9NRM6 , Q9Y6W8 , P05113 and P35225 , which represented nuocytes associated molecules were significantly increased in patients , meanwhile , the RORα mRNA also had a tendency to increase . In addition , IFN-γ and T-bet ( Th1 related cytokine and transcription factor ) were obviously decreased , and there was a positive correlation between Q9NRM6 and P35225 . These results suggested that there were polarized nuocytes in Graves ' hyperthyroidism patients , and which closely related to the down-regulation of Th1 cells or relatively advantage of Th2 differentiation . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Differential control of P10747 -regulated in vivo immunity by the E3 ligase Cbl-b . The E3 ubiquitin ligase Casitas B cell lymphoma-b ( Cbl-b ) plays a critical role in the development of autoimmunity and sets the threshold for T cell activation . In the absence of Cbl-b , T cells stimulated via the TCR respond similarly to those that have received a P10747 -mediated costimulatory signal , suggesting that the absence of Cbl-b substitutes for P10747 -mediated costimulation . In this study , we show that loss of Cbl-b restores Ig class switching and germinal center formation in Vav1 mutant mice in response to an in vivo viral challenge . Genetic inactivation of Cbl-b also rescues impaired antiviral IgG production in P10747 -mutant mice . Moreover , loss of P10747 results in disorganization of follicular dendritic cell clusters , which is also rescued by the Cbl-b mutation . Intriguingly , despite restored antiviral in vivo immunity and follicular dendritic cell clusters , loss of Cbl-b did not rescue germinal center formation in P10747 -deficient mice . Mechanistically , in vivo vesicular stomatitis virus-induced P05112 and P01579 production and up-regulation of the inducible costimulatory molecule Q9Y6W8 were dependent on P10747 , and could not be rescued by the loss of Cbl-b . These data provide genetic evidence that P10747 -dependent in vivo immune responses and Ig class switching can be genetically uncoupled from germinal center formation and Q9Y6W8 induction by Cbl-b-Vav1-regulated signaling pathways . Radiolabeled ligand binding to the catalytic or allosteric sites of O76074 and PDE11 . Cyclic nucleotide phosphodiesterases ( PDEs ) have been investigated for years as targets for therapeutic intervention in a number of pathophysiological processes . Phosphodiesterase-5 ( O76074 ) , which is highly specific for guanosine 3'-5'-cyclic-monophosphate ( cGMP ) at both its catalytic site and its allosteric sites , has generated particular interest because it is potently and specifically inhibited by three drugs : sildenafil ( Viagra , Pfizer ) , tadalafil ( DB00820 , Lilly- Q9Y6W8 ) , and vardenafil ( DB00862 , Bayer GSK ) . Previously , we have used [(3)H]cGMP to directly study the interaction of cGMP with the allosteric sites of O76074 , but because cGMP binds with relatively low affinity to the catalytic site , it has been difficult to devise a binding assay for this particular binding reaction . This approach using measurement of radiolabeled ligand binding continues to allow us to more precisely define functional features of the enzyme . We now use a similar approach to study the characteristics of high-affinity [(3)H]inhibitor binding to the O76074 catalytic domain . For these studies , we have prepared [(3)H]sildenafil and [(3)H]tadalafil , two structurally different competitive inhibitors of O76074 . The results demonstrate that radiolabeled ligands can be used as probes for both catalytic site and allosteric site functions of O76074 . We describe herein the methods that we have established for studying the binding of radiolabeled ligands to both types of sites on O76074 . These techniques have also been successfully applied to the study of binding of radiolabeled O76074 inhibitors to PDE11 , suggesting that these methods are applicable to the study of other PDEs , and perhaps other enzyme families . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs .

MH_train_7

MH_train_7

MH_train_7

interacts_with DB00773?

multiple_choice

Current researches on breast cancer epidemiology in Korea . As a cause of death in women , breast cancer ranks second to stomach cancer in Korea . Age-standardized mortality rates for breast cancer steadily increased during the 1980s and 1990s . There are big differences in the incidence rates for breast cancer compared with Western countries . Epidemiological features , trends in morbidity and mortality , various age-specific incidence curves , migrant study results , and analysis of the risk factors , however , suggest that the incidence of breast cancer might be further increasing in Korea . The key epidemiological hormonal risk factors for breast cancer are all explicable in terms of the estrogen augmented by progesterone hypothesis . These include older age , family history of breast cancer , early menarche , late menopause , late full-term pregnancy , and never a breast feeding . Both the establishment of high-risk groups and the estimation of lifetime risk are essential to develop a control strategy against breast cancer . Invasive ductal carcinoma is the most common histologic type of breast cancer in Korea , and the five-year survival rate has been estimated as 80-83 % . Recent studies on the identification of susceptibility factors such as genetic polymorphisms of P09488 /T1/P1 , P21964 , P05181 , P11511 , P05093 , P03372 , P18887 , O43542 , P43351 , TGF-alpha , P01375 , IL-1B , IL-1RN , P50613 etc. that predispose individuals to breast cancer by gene-environment or gene-gene interactions may possibly give further insight into both the etiology and the prevention of this malignancy . Levels of angiopoietins 1 and 2 in induced sputum supernatant in patients with P48444 . Pathological features of chronic obstructive pulmonary disease ( P48444 ) include lung vascular remodeling and angiogenesis . Q15389 ( Ang-1 ) , is an essential mediator of angiogenesis by establishing vascular integrity , whereas angiopoietin-2 ( Ang-2 ) acts as its natural inhibitor . We determined the levels of angiopoietins in sputum supernatants of patients with P48444 and investigated their possible association with mediators and cells involved in the inflammatory and remodeling process . Fifty-nine patients with P48444 , 25 healthy smokers and 20 healthy non-smokers were studied . All subjects underwent lung function tests , sputum induction for cell count identification and Ang-1 , Ang-2 , P15692 , TGF-β1 , P08253 , LTB4 , P10145 , albumin measurement in sputum supernatants . Airway vascular permeability ( AVP ) index was also assessed . Ang-2 levels were significantly higher in patients with P48444 compared to healthy smokers and healthy non-smokers [ median , interquartile ranges pg/ml , 267 ( 147-367 ) vs. 112 ( 67-171 ) and 98 ( 95-107 ) , respectively ; p < 0.001 ] . Regression analysis showed a significant association between Ang-2 levels and AVP index , P15692 , P10145 and P08253 levels in P48444 , the strongest being with P15692 . Our results indicate that induced sputum Ang-2 levels are higher in P48444 compared to healthy smokers and healthy non-smokers . Moreover , Ang-2 is associated with AVP , P10145 , P08253 , and P15692 , indicating a possible role for Ang-2 in the pathogenesis of the disease . P01308 -like growth factor-1 attenuates cisplatin-induced gammaH2AX formation and DNA double-strand breaks repair pathway in non-small cell lung cancer . Because insulin-like growth factor-1 ( DB01277 ) counteracts the anti-neoplastic effect of cisplatin that induces DNA damage and cell death through the formation of platinum-DNA adducts , we investigated the effects of DB01277 on the DNA double-strand breaks ( DSBs ) repair system induced by cisplatin . NCI-H1299 and H460 non-small cell lung cancer ( NSCLC ) cells treated with DB01277 recovered from cisplatin-derived inhibited proliferation and apoptosis . Decreased tail length in comet assay and suppressed phosphorylation of histone P16104 at Ser139 with DB01277 cotreatment indicates that DB01277 attenuates cisplatin-induced DNA damage . Cotreatment with DB01277 attenuates phosphorylation of ataxia-telangiectasia mutated ( Q13315 ) at Ser1981 , and Q13315 -Rad3-related ( ATR ) at Ser428 and subsequent phosphorylation of Chk2 , Chk1 , and p53 also dwindled by DB01277 . On the other hand , suppression of the IGF system with AG1024 or siRNA of insulin receptor substrate-1 ( P35568 ) , a major adaptor molecule of the IGF system , augmented cisplatin-induced gammaH2AX , Ser1981-pATM , and Ser428-pATR generation . Q13315 , which plays an important role in the phosphorylation of histone P16104 and Chk2 at Thr68 , strongly binds with P35568 under the influence of cisplatin , and the interaction was partially inhibited by DB01277 . Immunocytochemistry revealed that cisplatin induces nuclear translocation of P35568 with Ser1981-pATM , which is suppressed by cotreatment with DB01277 . In conclusion , cisplatin-induced gammaH2AX formation , DNA DSBs repair , and damage checkpoint pathway is inhibited by DB01277 . DB00515 derives interaction between Q13315 and P35568 , which is suppressed by DB01277 . Modulation of biologic activity of the DB01277 system could be a promising modality that raises the response rate of conventional chemotherapy . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment . 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 . Acidic pH induces topoisomerase II-mediated DNA damage . Acidic pH plays an important role in various pathophysiological states and has been demonstrated to be carcinogenic in animal models . Recent studies have also implicated acidic pH in the development of preneoplastic Barrett 's esophagus in human . However , little is known about the molecular mechanism underlying acidic pH-induced carcinogenesis . In the current study , we show that acidic pH , like the topoisomerase II ( P11388 ) poison DB00773 ( demethylepipodophyllotoxin ethylidene-beta-D-glucoside ) , induces tumors in 9,10-dimethyl-1,2-benzanthracene(DMBA)-initiated mice . The following studies in tissue culture models have suggested that acidic pH acts like a P11388 poison to induce P11388 -mediated DNA damage : ( i ) acidic pH induces P11388 -dependent DNA damage signals as evidenced by up-regulation of p53 and DB00133 -139 phosphorylation of P16104 [ a substrate for ataxia telangiectasia mutated ( Q13315 ) Q13315 and Rad3-related ( ATR ) kinases ] ; ( ii ) acidic pH-induced cytotoxicity in tumor cells is reduced in P11388 -deficient cells ; ( iii ) acidic pH increases the mutation frequency of the hypoxanthine phosphoribosyl transferase ( P00492 ) gene in a P11388 -dependent manner ; and ( iv ) acidic pH induces reversible P11388 -mediated DNA strand breaks in vitro . We discuss the possibility that P11388 -mediated DNA damage may contribute to acidic pH-induced carcinogenesis . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Pharmacogenetics of oral antidiabetic drugs . Oral antidiabetic drugs ( OADs ) are used for more than a half-century in the treatment of type 2 diabetes . Only in the last five years , intensive research has been conducted in the pharmacogenetics of these drugs based mainly on the retrospective register studies , but only a handful of associations detected in these studies were replicated . The gene variants in P11712 , Q09428 / Q14654 , and Q9NQB0 were associated with the effect of sulfonylureas . P11712 encodes sulfonylurea metabolizing cytochrome P450 isoenzyme 2C9 , Q09428 and Q14654 genes encode proteins constituting DB00171 -sensitive K(+) channel which is a therapeutic target for sulfonylureas , and Q9NQB0 is a gene with the strongest association with type 2 diabetes . O15245 , Q96FL8 , and Q13315 gene variants were repeatedly associated with the response to metformin . O15245 and Q96FL8 encode metformin transporters OCT1 and Q96FL8 , respectively . The function of a gene variant near Q13315 gene identified by a genome-wide association study is not elucidated so far . The first variant associated with the response to gliptins is a polymorphism in the proximity of P17538 /2 gene which encodes chymotrypsinogen . Establishment of diabetes pharmacogenetics consortia and reduction in costs of genomics might lead to some significant clinical breakthroughs in this field in a near future . The Escherichia coli recA gene increases resistance of the yeast Saccharomyces cerevisiae to ionizing and ultraviolet radiation . The Escherichia coli recA protein coding region was ligated into an extrachromosomally replicating yeast expression vector downstream of the yeast alcohol dehydrogenase promoter region to produce plasmid pADHrecA . Transformation of the wild-type yeast strains YNN-27 and 7799-4B , as well as the recombination-deficient rad52-1 P01031 -6 mutant , with this shuttle plasmid resulted in the expression of the bacterial 38 kDa RecA protein in exponential phase cells . The wild-type YNN27 and 7799-4B transformants expressing the bacterial recA gene showed increased resistance to the toxic effects of both ionizing and ultraviolet radiation . RecA moderately stimulated the UV-induced mutagenic response of 7799-4B cells . Transformation of the rad52-1 mutant with plasmid pADHrecA did not result in the complementation of sensitivity to ionizing radiation . Thus , the RecA protein endows the yeast cells with additional activities , which were shown to be error-prone and dependent on the P43351 gene . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Cigarette smoke induces cellular senescence . Chronic obstructive pulmonary disease ( P48444 ) is the fourth leading cause of death in the United States , and cigarette smoking is the major risk factor for P48444 . Fibroblasts play an important role in repair and lung homeostasis . Recent studies have demonstrated a reduced growth rate for lung fibroblasts in patients with P48444 . In this study we examined the effect of cigarette smoke extract ( CSE ) on fibroblast proliferative capacity . We found that cigarette smoke stopped proliferation of lung fibroblasts and upregulated two pathways linked to cell senescence ( a biological process associated with cell longevity and an inability to replicate ) , p53 and p16-retinoblastoma protein pathways . We compared a single exposure of CSE to multiple exposures over an extended time course . A single exposure to CSE led to cell growth inhibition at multiple phases of the cell cycle without killing the cells . The decrease in proliferation was accompanied by increased Q13315 , p53 , and P38936 activity . However , several important senescent markers were not present in the cells at an earlier time point . When we examined multiple exposures to CSE , we found that the cells had profound growth arrest , a flat and enlarged morphology , upregulated p16 , and senescence-associated beta-galactosidase activity , which is consistent with a classic senescent phenotype . These observations suggest that while a single exposure to cigarette smoke inhibits normal fibroblast proliferation ( required for lung repair ) , multiple exposures to cigarette smoke move cells into an irreversible state of senescence . This inability to repair lung injury may be an essential feature of emphysema . Topoisomerase II-mediated DNA cleavage and mutagenesis activated by nitric oxide underlie the inflammation-associated tumorigenesis . AIMS : Both cancer-suppressing and cancer-promoting properties of reactive nitrogen and oxygen species ( RNOS ) have been suggested to play a role in tumor pathology , particularly those activities associated with chronic inflammation . Here , we address the impact of nitric oxide ( NO ) on the induction of DNA damage and genome instability with a specific focus on the involvement of topoisomerase II ( P11388 ) . We also investigate the contribution of NO to the formation of skin melanoma in mice . RESULTS : Similar to the P11388 -targeting drug , etoposide ( DB00773 ) , the NO-donor , S-nitrosoglutathione ( GSNO ) , induces skin melanomas formation in 7,12-dimethyl- benz[a]anthracene ( DMBA ) -initiated mice . To explore the mechanism(s) underlying this NO-induced tumorigenesis , we use a co-culture model system to demonstrate that inflamed macrophages with inducible NO synthase ( P35228 ) expression cause γ- P16104 activation , p53 phosphorylation , and chromosome DNA breaks in the target cells . Inhibitor experiments revealed that NO and P11388 isozymes are responsible for the above described cellular phenotypes . Notably , NO , unlike DB00773 , preferentially induces the formation of TOP2β cleavable complexes ( TOP2βcc ) in cells . Moreover , GSNO induced P11388 -dependent DNA sequence rearrangements and cytotoxicity . Furthermore , the incidences of GSNO- and DB00773 -induced skin melanomas were also observed to be lower in the skin-specific top2β-knockout mice . Our results suggest that P11388 isozymes contribute to NO-induced mutagenesis and subsequent cancer development during chronic inflammation . INNOVATION AND CONCLUSIONS : We provide the first experimental evidence for the functional role of P11388 in NO-caused DNA damage , mutagenesis , and carcinogenesis . Notably , these studies contribute to our molecular understanding of the cancer-promoting actions of RNOS during chronic inflammation . Loss of P38398 function increases the antitumor activity of cisplatin against human breast cancer xenografts in vivo . BACKGROUND : Previous reports suggested a central role of P38398 in DNA-damage repair mechanisms elicited by cell exposure to anti-tumor agents . Here we studied if P38398 -defective HCC1937 or P38398 -reconstituted HCC1937/(WT) P38398 human breast cancer xenografts ( HBCXs ) generated in SCID mice were differentially sensitive to cisplatin ( DB00515 ) in vivo and we investigated potential molecular correlates of this effect . RESULTS : DB00515 induced almost complete growth inhibition of P38398 -defective HBCXs , while P38398 -reconstituted HBCXs were only partially inhibited . Cell cycle analysis showed a significant S- and G(2)/M blockade in P38398 -defective as compared with parental P38398 -reconstituted cells . Comparative gene expression profiling of HCC1937 and HCC1937/(WT) P38398 showed upregulation of P43351 and Q13426 , whereas P07992 and P23921 were downregulated . Pathway finder analysis of gene arrays data indicated perturbations of major proliferation and survival pathways suggesting that P38398 is mostly involved in G(2)/M but also in G(1)/S-phase checkpoints as well as in several important signaling pathways , including IGF , P15692 , estrogen receptor , PI3K/AKT and P01133 . METHODS : HCC1937 or HCC1937/(WT) P38398 HBCXs were generated in SCID mice . Animals were then weekly treated with 5 mg/kg DB00515 i.p. or with vehicle for 4 w . Tumor volume and mice survival were evaluated . Tumors were retrieved from animals 12 hours after the last treatment with DB00515 or vehicle treatment and the cell suspension underwent cell cycle analysis . Differential gene expression and pathway modulation between HCC1937 and HCC1937/(WT) P38398 cells were also studied . CONCLUSION : Our data suggest that P38398 -defective in vivo HBCXs express a molecular scenario predictive of high sensitivity to platinum-derived compounds strongly supporting the rationale for prospective tailored clinical trials in hereditary breast cancer . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Identification and validation of commonly overexpressed genes in solid tumors by comparison of microarray data . Cancers originating from epithelial cells are the most common malignancies . No common expression profile of solid tumors compared to normal tissues has been described so far . Therefore we were interested if genes differentially expressed in the majority of carcinomas could be identified using bioinformatic methods . Complete data sets were downloaded for carcinomas of the prostate , breast , lung , ovary , colon , pancreas , stomach , bladder , liver , and kidney , and were subjected to an expression analysis using DB00118 . In each experiment , a gene was scored as differentially expressed if the q value was below 25 % . Probe identifiers were unified by comparing the respective probe sequences to the Unigene build 155 using BlastN . To obtain differentially expressed genes within the set of analyzed carcinomas , the number of experiments in which differential expression was observed was counted . Differential expression was assigned to genes if they were differentially expressed in at least eight experiments of tumors from different origin . The identified candidate genes Q16186 , Q99848 , P14324 , Q08050 , P16104 , O15379 , P51617 , and P25490 were subjected to further validation . Using this comparative approach , 100 genes were identified as upregulated and 21 genes as downregulated in the carcinomas . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Direct interaction of Q9BXW9 with P51587 in DNA damage response pathways . Fanconi anaemia ( FA ) is a chromosomal instability disorder characterized by cellular sensitivity to DNA interstrand crosslinking agents and a high risk of cancer . Six of the eight proteins encoded by the known FA genes form a nuclear complex which is required for the monoubiquitination of the Q9BXW9 protein . Q9BXW9 complexes and colocalizes with P38398 , but its presumptive role in DNA repair has not yet been clearly defined . We used yeast two-hybrid analysis to test for interaction between Q9BXW9 and 10 proteins involved in homologous recombination repair . Q9BXW9 did not interact with Q06609 , the five Q06609 paralogs , P43351 , RAD54 or DMC1 . However , it bound to a highly conserved C-terminal site in P51587 that also binds O15287 / O15287 . Q9BXW9 and P51587 can be coimmunoprecipitated from cell extracts of both human and Chinese hamster wild-type cells , thus confirming that the interaction occurs in vivo . Formation of nuclear foci of Q9BXW9 was normal in the P51587 mutant CAPAN-1 cells , which indicates that the recruitment of Q9BXW9 to sites of DNA-repair is independent of wild-type P51587 function . Q9BXW9 colocalized with Q06609 in foci following treatment with mitomycin C or hydroxyurea , and colocalized very tightly with P12004 after treatment with hydroxyurea . These findings suggest that Q9BXW9 may have a role in the cellular response to stalled replication forks or in the repair of replication-associated double-strand breaks , irrespective of the type of primary DNA lesion . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Resistance of glioblastoma-initiating cells to radiation mediated by the tumor microenvironment can be abolished by inhibiting transforming growth factor-β . The poor prognosis of glioblastoma ( GBM ) routinely treated with ionizing radiation ( IR ) has been attributed to the relative radioresistance of glioma-initiating cells ( GIC ) . Other studies indicate that although GIC are sensitive , the response is mediated by undefined factors in the microenvironment . GBM produce abundant transforming growth factor-β ( TGF-β ) , a pleotropic cytokine that promotes effective DNA damage response . Consistent with this , radiation sensitivity , as measured by clonogenic assay of cultured murine ( GL261 ) and human ( U251 , U87MG ) glioma cell lines , increased by approximately 25 % when treated with LY364947 , a small-molecule inhibitor of TGF-β type I receptor kinase , before irradiation . Mice bearing GL261 flank tumors treated with 1D11 , a pan-isoform TGF-β neutralizing antibody , exhibited significantly increased tumor growth delay following IR . GL261 neurosphere cultures were used to evaluate GIC . LY364947 had no effect on the primary or secondary neurosphere-forming capacity . IR decreased primary neurosphere formation by 28 % , but did not reduce secondary neurosphere formation . In contrast , LY364947 treatment before IR decreased primary neurosphere formation by 75 % and secondary neurosphere formation by 68 % . Notably , GL261 neurospheres produced 3.7-fold more TGF-β per cell compared with conventional culture , suggesting that TGF-β production by GIC promotes effective DNA damage response and self-renewal , which creates microenvironment-mediated resistance . Consistent with this , LY364947 treatment in irradiated GL261 neurosphere-derived cells decreased DNA damage responses , P16104 and p53 phosphorylation , and induction of self-renewal signals , Notch1 and P61073 . These data motivate the use of TGF-β inhibitors with radiation to improve therapeutic response in patients with GBM . DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . Chk2-dependent phosphorylation of P18887 in the DNA damage response promotes base excision repair . The DNA damage response ( DDR ) has an essential function in maintaining genomic stability . Ataxia telangiectasia-mutated ( Q13315 ) -checkpoint kinase 2 ( Chk2 ) and Q13315 - and Rad3-related ( ATR ) -Chk1 , triggered , respectively , by DNA double-strand breaks and blocked replication forks , are two major DDRs processing structurally complicated DNA damage . In contrast , damage repaired by base excision repair ( BER ) is structurally simple , but whether , and how , the DDR is involved in repairing this damage is unclear . Here , we demonstrated that Q13315 -Chk2 was activated in the early response to oxidative and alkylation damage , known to be repaired by BER . Furthermore , Chk2 formed a complex with P18887 , the BER scaffold protein , and phosphorylated P18887 in vivo and in vitro at DB00156 (284) . A mutated P18887 lacking DB00156 (284) phosphorylation was linked to increased accumulation of unrepaired BER intermediate , reduced DNA repair capacity , and higher sensitivity to alkylation damage . In addition , a phosphorylation-mimic form of P18887 showed increased interaction with glycosylases , but not other BER proteins . Our results are consistent with the phosphorylation of P18887 by Q13315 -Chk2 facilitating recruitment of downstream BER proteins to the initial damage recognition/excision step to promote BER . Increased expression of human DNA repair genes , P18887 , O43542 and Q06609 , in radioresistant human KB carcinoma cell line N10 . The radioresistant N10 and parental KB cell lines were examined for the expression of human DNA repair genes which were related to the repair of radiation-induced DNA damage by northern blot analysis using five kinds of DNA probes ( P18887 , O43542 , P13010 , Q06609 , P43351 ) . In the unirradiated condition , N10 cells showed higher expression of P18887 , O43542 and Q06609 mRNA than did KB cells . The X-irradiation induced a time-dependent increase in the mRNA levels of O43542 and Q06609 in both cell lines with a maximum at 2 h postirradiation . The P18887 mRNA in N10 was maintained at the same level even after irradiation , whereas that in KB was decreased after irradiation . There was no difference in the expression of P13010 and P43351 mRNA between N10 and KB cells in both unirradiated and irradiated conditions . From these findings , it was suggested that P18887 , O43542 and Q06609 contribute to the radioresistance in cell line N10 . Pharmacogenetics in chemotherapy of colorectal cancer . Although in recent years , chemotherapeutic options for colorectal carcinoma have expanded , overall response rates are still too low , with high rates of toxicity . Pharmacogenetics aim at predicting both treatment response and adverse effects in individual patients . This review describes the current knowledge of pharmacogenetic markers in the systemic treatment of colorectal cancer . P22309 *28 leads to reduced conjugation of SN-38 , the active metabolite of irinotecan , resulting in an increased rate of adverse effects , especially neutropenia . To a lesser extent , increased DB00544 toxicity is predicted by Q12882 *2A . A variable number of tandem repeats polymorphism in the thymidylate synthase enhancer region , in combination with a single nucleotide polymorphism C > G , may predict poorer response to DB00544 . Efficacy of oxaliplatin is influenced by polymorphisms in components of DNA repair systems , such as P07992 and P18887 . Polymorphic changes in the endothelial growth factor receptor probably predict cetuximab efficacy . Furthermore , the antibody-depended cell-mediated cytotoxic effect of cetuximab may be reduced by polymorphisms in the immunoglobin G fragment C receptors . DB00112 efficacy is suspected to be influenced by polymorphisms in the P15692 gene and the hypoxia inducible factor 1alpha gene . Although the interpretation of pharmacogenetic studies is complicated , results imply a promising way of pretreatment prediction of chemotherapy efficacy and toxicity . Oxidative DNA damage in lung tissue from patients with P48444 is clustered in functionally significant sequences . Lung tissue from P48444 patients displays oxidative DNA damage . The present study determined whether oxidative DNA damage was randomly distributed or whether it was localized in specific sequences in either the nuclear or mitochondrial genomes . The DNA damage-specific histone , gamma- P16104 , was detected immunohistochemically in alveolar wall cells in lung tissue from P48444 patients but not control subjects . A PCR-based method was used to search for oxidized purine base products in selected 200 bp sequences in promoters and coding regions of the P15692 , TGF-β1 , P09601 , Egr1 , and β-actin genes while quantitative Southern blot analysis was used to detect oxidative damage to the mitochondrial genome in lung tissue from control subjects and P48444 patients . Among the nuclear genes examined , oxidative damage was detected in only 1 sequence in lung tissue from P48444 patients : the hypoxic response element ( HRE ) of the P15692 promoter . The content of P15692 mRNA also was reduced in P48444 lung tissue . Mitochondrial DNA content was unaltered in P48444 lung tissue , but there was a substantial increase in mitochondrial DNA strand breaks and/or abasic sites . These findings show that oxidative DNA damage in P48444 lungs is prominent in the HRE of the P15692 promoter and in the mitochondrial genome and raise the intriguing possibility that genome and sequence-specific oxidative DNA damage could contribute to transcriptional dysregulation and cell fate decisions in P48444 . Farnesyl diphosphate synthase : the art of compromise between substrate selectivity and stereoselectivity . Farnesyl diphosphate ( FPP ) synthase catalyzes the consecutive head-to-tail condensations of isopentenyl diphosphate ( IPP , P01031 ) with dimethylallyl diphosphate ( DMAPP , P01031 ) and geranyl diphosphate ( GPP , Q99622 ) to give ( E,E ) -FPP ( C15 ) . The enzyme belongs to a genetically distinct family of chain elongation enzymes that install E-double bonds during each addition of a five-carbon isoprene unit . Analysis of the Q99622 and C15 products from incubations with avian P14324 reveals that small amounts of neryl diphosphate ( Z- Q99622 ) and ( Z,E ) -FPP are formed along with the E-isomers during the P01031 --> Q99622 and Q99622 --> C15 reactions . Similar results were obtained for P14324 from Escherichia coli , Artemisia tridentata ( sage brush ) , Pyrococcus furiosus , and Methanobacter thermautotrophicus and for GPP and FPP synthesized in vivo by E. coli P14324 . When ( R ) -[2-2H]IPP was a substrate for chain elongation , no deuterium was found in the chain elongation products . In contrast , the deuterium in ( S ) -[2-2H]IPP was incorporated into all of the products . Thus , the pro-R hydrogen at P06681 of IPP is lost when the E- and Z-double bond isomers are formed . The synthesis of Z-double bond isomers by P14324 during chain elongation is unexpected for a highly evolved enzyme and probably reflects a compromise between optimizing double bond stereoselectivity and the need to exclude DMAPP from the IPP binding site . 15-deoxy-Δ¹²,¹⁴-PGJ₂ promotes inflammation and apoptosis in cardiomyocytes via the Q14188 /MAPK/ P01375 α axis . BACKGROUND : Prostaglandins ( PGs ) , lipid autacoids derived from arachidonic acid , play a pivotal role during inflammation . P52209 ₂ synthase is abundantly expressed in heart tissue and P52209 ₂ has recently been found to induce cardiomyocyte apoptosis . P52209 ₂ is an unstable prostanoid metabolite ; therefore the objective of the present study was to elucidate whether its final dehydration product , 15-deoxy-Δ¹²,¹⁴-PGJ₂ ( 15d-PGJ₂ , present at high levels in ischemic myocardium ) might cause cardiomyocyte damage . METHODS AND RESULTS : Using specific (ant)agonists we show that 15d-PGJ₂ induced formation of intracellular reactive oxygen species ( ROS ) and phosphorylation of p38 and Q8NFH3 /44 MAPKs via the Q13258 Q14188 ( but not DP1 or Q07869 γ ) in the murine atrial cardiomyocyte P07306 cell line . Activation of the Q14188 -ROS-MAPK axis by 15d-PGJ₂ enhanced transcription and translation of P01375 α and induced apoptosis in P07306 cardiomyocytes . Silencing of P01375 α significantly attenuated the extrinsic ( caspase-8 ) and intrinsic apoptotic pathways ( bax and caspase-9 ) , caspase-3 activation and downstream PARP cleavage and γ P16104 activation . The apoptotic machinery was unaffected by intracellular calcium , transcription factor NF-κB and its downstream target p53 . Of note , 9,10-dihydro-15d-PGJ₂ ( lacking the electrophilic carbon atom in the cyclopentenone ring ) did not activate cellular responses . Selected experiments performed in primary murine cardiomyocytes confirmed data obtained in P07306 cells namely that the intrinsic and extrinsic apoptotic cascades are activated via Q14188 /MAPK/ P01375 α signaling . CONCLUSIONS : We conclude that the reactive α,β-unsaturated carbonyl group of 15d-PGJ₂ is responsible for the pronounced upregulation of P01375 α promoting cardiomyocyte apoptosis . We propose that inhibition of Q14188 receptors could provide a possibility to modulate 15d-PGJ₂-induced myocardial injury . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . Growth of V79 cells as xenograft tumors promotes multicellular resistance but does not increase spontaneous or radiation-induced mutant frequency . A Chinese hamster V79 xenograft model was developed to determine whether cells subjected to a hypoxic tumor microenvironment would be more likely to undergo mutation at the P00492 locus . V79-171b cells stably transfected with P15692 and EGFP were grown subcutaneously in immunodeficient NOD/ SCID mice . V79-VE tumors were characterized for host cell infiltration , doubling time , hypoxic fraction , vascular perfusion , and response to ionizing radiation . When irradiated in vitro , the mutant frequency for a given surviving fraction did not differ for cells grown in vivo or in vitro . Similar results were obtained using HCT116 human colorectal carcinoma cells grown as xenografts . However , V79-VE cells grown as xenografts were significantly more resistant to killing than monolayers . The background mutant frequency and the radiation-induced mutant frequency did not differ for tumor cells close to or distant from blood vessels . Similarly , tumor cells from well-perfused regions showed the same rate of strand break rejoining and the same rate of loss of phosphorylated histone P16104 as cells sorted from poorly perfused regions . Therefore , deleterious effects of the tumor microenvironment on DNA repair efficiency or mutation induction could not be demonstrated in these tumors . Rather , development of multicellular resistance in V79-VE tumors acted to reduce mutant frequency for a given dose of radiation . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . Clinicopathological and functional significance of P18887 expression in ovarian cancer . X-ray repair cross-complementing gene 1 ( P18887 ) is essential for DNA base excision repair , single strand break repair and nucleotide excision repair . We investigated clinicopathological and functional significance of P18887 expression in ovarian cancers . P18887 protein expression was evaluated in 195 consecutive human ovarian cancers and correlated with clinicopathological variables and survival outcomes . Functional preclinical studies were conducted in a panel of P18887 deficient and proficient Chinese hamster and Human cancer cells for cisplatin chemosensitivity . Clonogenic assay , neutral COMET assay , γ P16104 immunocytochemistry and flow cytometric analyses were performed in cells . In ovarian cancer , 48 % of the tumors were positive for P18887 expression and significantly associated with higher stage ( p = 0.006 ) , serous type tumors ( p = 0.008 ) , suboptimal de-bulking ( p = 0.004 ) and platinum resistance ( p < 0.0001 ) . Positive P18887 had twofold increase of risk of death ( p = 0.007 ) and progression ( p < 0.0001 ) . In the multivariate Cox model , P18887 expression was independently associated with cancer specific [ p = 0.038 ] and progression free survival [ p = 0.003 ] . Preclinically , P18887 negative cells were sensitive to cisplatin compared to P18887 positive cells . Sensitivity to cisplatin in P18887 negative cells was associated with accumulation of DNA double strand breaks and G2/M cell cycle arrest . P18887 expression is associated with adverse clinicopathological and survival outcomes in patients . Preclinical data provides mechanistic functional evidence for cisplatin sensitivity in P18887 negative cells . P18887 is a promising predictive biomarker in ovarian cancer . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Mitoxantrone inhibits HIF-1α expression in a topoisomerase II-independent pathway . PURPOSE : Solid tumors encounter a growth-limiting hypoxic microenvironment as they develop . Hypoxia-inducible factors ( HIF ) play important roles in hypoxia-associated tumor development and therapeutic resistance . Targeting the HIF pathway ( especially HIF-1α ) represents a promising cancer treatment strategy . Here , we report a novel class of HIF-1α inhibitors and the possible molecular basis of inhibition . EXPERIMENTAL DESIGN : We analyzed the inhibitory effects of clinically used topoisomerase II ( P11388 ) -targeting drugs on HIF-1α expression with a primary focus on mitoxantrone . The potential role of P11388 in mitoxantrone-inhibited HIF-1α expression was studied using pharmacologic inhibition , a knockdown approach , and P11388 mutant cells . Moreover , involvement of mitoxantrone in proteasome-mediated degradation , transcription , and translation of HIF-1α was examined . RESULTS : The P11388 -targeting mitoxantrone , but neither doxorubicin nor etoposide ( DB00773 ) , strongly inhibited HIF-1α expression under hypoxic conditions in a dose- and time-dependent manner . Surprisingly , the mitoxantrone-mediated inhibition of HIF-1α expression was largely independent of two P11388 isozymes , proteasomal degradation , and transcription . Furthermore , mitoxantrone inhibited HIF-1α expression and function in a similar fashion as cycloheximide , suggesting that mitoxantrone might inhibit HIF-1α via a blockage at its translation step . In vitro translation experiments using HIF-1α mRNA further confirmed inhibition of HIF-1α translation by mitoxantrone . Interestingly , levels of the polysome-bound HIF-1α and P15692 mRNA were elevated and decreased after mitoxantrone treatment , respectively . CONCLUSIONS : We have identified the P11388 -targeting compound , mitoxantrone , as an HIF-1α inhibitor possibly through a translation inhibition mechanism , suggesting the possibility of an additional anticancer activity for mitoxantrone . TGF-β1-ROS- Q13315 -CREB signaling axis in macrophage mediated migration of human breast cancer MCF7 cells . Macrophages in the tumor microenvironment play an important role in tumor cell survival . They influence the tumor cell to proliferate , invade into surrounding normal tissues and metastasize to local and distant sites . In this study , we evaluated the effect of conditioned medium from monocytes and macrophages on growth and migration of breast cancer cells . Macrophage conditioned medium ( MϕCM ) containing elevated levels of cytokines P01375 -α , IL-1β and P05231 had a differential effect on non-invasive ( MCF7 ) and highly invasive ( MDA-MB-231 ) breast cancer cell lines . MϕCM induced the secretion of TGF-β1 in MCF7 cells . This was associated with apoptosis in a fraction of cells and generation of reactive oxygen and nitrogen species ( ROS and RNS ) and DNA damage in the remaining cells . This , in turn , increased expression of DB02527 response element binding protein ( CREB ) and vimentin resulting in migration of cells . These effects were inhibited by neutralization of P01375 -α , IL-1β and P05231 , inhibition of ROS and RNS , DNA damage and siRNA mediated knockdown of Q13315 . In contrast , MDA-MB-231 cells which had higher basal levels of pCREB were not affected by MϕCM . In summary , we have found that pro-inflammatory cytokines secreted by macrophages induce TGF-β1 in tumor cells , which activate pCREB signaling , epithelial-mesenchymal-transition ( EMT ) responses and enhanced migration . mRNA expression , functional profiling and multivariate classification of colon biopsy specimen by cDNA overall glass microarray . AIM : To understand the local pathophysiological alterations and gene ontology-based functional classification of colonic biopsies into inflammatory and neoplastic diseases . METHODS : Total RNA was extracted from frozen biopsies and amplified by T7-method . Expression profile was evaluated by Atlas Glass 1K microarrays . After microarray quality control , applicable data were available from 10 adenomas , 6 colorectal adenocarcinomas ( CRCs ) , and 6 inflammatory bowel diseases ( IBDs ) . Multivariate statistical and cell functional analyses were performed . Real-time RT-PCR and immunohistochemistry were used for validation . RESULTS : Discriminant analysis of selected genes , could correctly reclassify all 22 samples using 4 parameters ( heat shock transcription factor-1 , bystin-like , calgranulin-A , O14798 ) . Q9UKU7 samples were characterized by overregulated chemokine ( C-X-C motif ) ligand 13 , replication protein A1 , Q15723 and downregulated Q9Y4K3 , P10415 -interacting killer genes . In adenomas upregulation of Q9Y4K3 , replication protein A1 , Q15723 and underexpression of P10415 -associated X protein , calgranulin-A genes were found . CRC cases had significantly increased epidermal growth factor receptor , topoisomerase-1 , v-jun , Q9Y4K3 and O14798 , and decreased Q06609 and P43351 DNA repair gene , protein phosphatase-2A and P10415 -interacting killer mRNA levels . P00533 RT-PCR and immunohistochemistry , topoisomerase-1 RT-PCR confirmed the chip results . CONCLUSION : Different histological alterations can be reclassified by functional , multivariate analysis using cDNA microarrays . Further studies with expanded sample number are needed for subclassification of pathological alterations . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . Induction of apoptosis of Beta cells of the pancreas by advanced glycation end-products , important mediators of chronic complications of diabetes mellitus . We herein report cytotoxicity of advanced glycation end-products ( AGEs ) on pancreatic beta cells . AGEs stimulated reactive oxygen species ( ROS ) generation but did not arrest proliferation of the P01308 -1 cell line . Pancreatic beta cell lines or primary cultured islets possess a receptor for P51606 ( RAGE ) , and its expression increased after P51606 treatment . TUNEL staining and FACS analysis using annexin V/PI antibodies showed that apoptosis increased in P01308 -1 cells or primary cultured islets when incubated with BSA conjugated with glyceraldehyde ( AGE2 ) or glucoaldehyde ( AGE3 ) , compared with those conjugated with glucose ( AGE1 ) . Reaction of P01308 -1 cells to Ki67 , which is a cellular marker for proliferation , was also increased after P51606 treatment . The ability of primary cultured islets to secrete insulin was retained even after P51606 treatment under either low or high glucose conditions . The antiserum against RAGE partially prevented P51606 -induced cellular events . Treatment of beta cells with the antioxidant metallothionein results in a significant reduction in pathologic changes . AGEs might be able to induce apoptosis as well as proliferation of pancreatic beta cell lines or primary cultured islets . Moreover , antibody array showed that Q06609 and P43351 were significantly decreased in AGE2-treated P01308 -1 cells . AGEs might inhibit homologous DNA recombination for repairing DNA of P01308 -1 cells damaged by ROS generation . It might be suggested that treatment of AGEs resulted in ROS production and apoptosis through their receptor on pancreatic beta cells . AGEs might deteriorate function of pancreatic beta cells in patients with long-term hyperglycemia . Ablation of cholesterol biosynthesis in neural stem cells increases their P15692 expression and angiogenesis but causes neuron apoptosis . Although sufficient cholesterol supply is known to be crucial for neurons in the developing mammalian brain , the cholesterol requirement of neural stem and progenitor cells in the embryonic central nervous system has not been addressed . Here we have conditionally ablated the activity of squalene synthase ( P37268 ) , a key enzyme for endogenous cholesterol production , in the neural stem and progenitor cells of the ventricular zone ( VZ ) of the embryonic mouse brain . Mutant embryos exhibited a reduced brain size due to the atrophy of the neuronal layers , and died at birth . Analyses of the E11.5-E15.5 dorsal telencephalon and diencephalon revealed that this atrophy was due to massive apoptosis of newborn neurons , implying that this progeny of the P37268 -ablated neural stem and progenitor cells was dependent on endogenous cholesterol biosynthesis for survival . Interestingly , the neural stem and progenitor cells of the VZ , the primary target of P37268 inactivation , did not undergo significant apoptosis . Instead , vascular endothelial growth factor ( P15692 ) expression in these cells was strongly upregulated via a hypoxia-inducible factor-1-independent pathway , and angiogenesis in the VZ was increased . Consistent with an increased supply of lipoproteins to these cells , the level of lipid droplets containing triacylglycerides with unsaturated fatty acyl chains was found to be elevated . Our study establishes a direct link between intracellular cholesterol levels , P15692 expression , and angiogenesis . Moreover , our data reveal a hitherto unknown compensatory process by which the neural stem and progenitor cells of the developing mammalian brain evade the detrimental consequences of impaired endogenous cholesterol biosynthesis . Dynamic coregulatory complex containing P38398 , Q01094 and Q99708 controls Q13315 transcription . Chromosomal instability is a key feature in cancer progression . Recently we have reported that P38398 regulates the transcription of several genes in prostate cancer , including Q13315 ( ataxia telangiectasia mutated ) . Although it is well accepted that Q13315 is a pivotal mediator in genotoxic stress , it is unknown whether Q13315 transcription is regulated during the molecular response to DNA damage . Here we investigate Q13315 transcription regulation in human prostate tumor PC3 cell line . We have found that doxorubicin and mitoxantrone repress Q13315 transcription in PC3 cells but etoposide and methotrexate do not affect Q13315 expression . We have demonstrated that P38398 binds to Q13315 promoter and after doxorubicin exposure , it is released . P38398 overexpression increases Q13315 transcription and this enhancement is abolished by P38398 depletion . Moreover , P38398 -BRCT domain loss impairs the ability of P38398 to regulate Q13315 promoter activity , strongly suggesting that BRCT domain is essential for Q13315 regulation by P38398 . P38398 -overexpressing PC3 cells exposed to KU55933 Q13315 kinase inhibitor showed significant decreased Q13315 promoter activity compared to untreated cells , suggesting that Q13315 transcriptional regulation by P38398 is partially mediated by the Q13315 kinase activity . In addition , we have demonstrated Q01094 binding to Q13315 promoter before and after doxorubicin exposure . Q01094 overexpression diminishes Q13315 transcription after doxorubicin exposure which is impaired by Q01094 dominant negative mutants . Finally , the co-regulator of transcription Q99708 increases Q13315 transcription . Q99708 increases Q13315 transcription . Altogether , P38398 / Q01094 / Q99708 binding to Q13315 promoter activates Q13315 transcription . Doxorubicin exposure releases P38398 and Q99708 from Q13315 promoter still keeping Q01094 recruited and , in turn , represses Q13315 expression . Identification of the fused bicyclic 4-amino-2-phenylpyrimidine derivatives as novel and potent DB05876 inhibitors . 2-Phenyl-4-piperidinyl-6,7-dihydrothieno[3,4-d]pyrimidine derivative ( 2 ) was found to be a new DB05876 inhibitor with moderate Q07343 activity ( IC50=150 nM ) . A number of derivatives with a variety of 4-amino substituents and fused bicyclic pyrimidines were synthesized . Among these , 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivative ( 18 ) showed potent Q07343 inhibitory activity ( IC50=25 nM ) . Finally , N-propylacetamide derivative ( 31b ) was determined as a potent inhibitor for both Q07343 ( IC50=7.5 nM ) and P01375 -α production in mouse splenocytes ( IC50=9.8 nM ) and showed good in vivo anti-inflammatory activity in the LPS-induced lung inflammation model in mice ( ID50=18 mg/kg ) . The binding mode of the new inhibitor ( 31e ) in the catalytic site of Q07343 is presented based on an X-ray crystal structure of the ligand-enzyme complex . Amsacrine and etoposide hypersensitivity of yeast cells overexpressing DNA topoisomerase II . Increasing the cellular concentration of DNA topoisomerase II in yeast by expressing constitutively a plasmid-borne P11388 gene encoding the enzyme greatly increases the sensitivity of the cells to amsacrine and etoposide ( DB00773 ) . This increased drug sensitivity at a higher intracellular DNA topoisomerase II level is observed in both P43351 + repair-proficient strains and rad52 mutants that are defective in the repair of double-stranded breaks . These results provide strong support of the hypothesis that the cellular target of these drugs is DNA topoisomerase II , and that these drugs kill cells by converting DNA topoisomerase II into a DNA damaging agent . P01308 secretory defects and impaired islet architecture in pancreatic beta-cell-specific P40763 knockout mice . Normal islet formation and function depends on the action of various growth factors operating in pre- and postnatal development ; however , the specific physiological function of each factor is largely unknown . Loss-of-function analyses in mice have provided little information so far , perhaps due to functional redundancies of the growth factors acting on the pancreas . The present study focuses on the role of the transcription factor P40763 in insulin-producing cells . P40763 is one of the potential downstream mediators for multiple growth factors acting on the pancreatic beta-cells , including betacellulin , hepatocyte growth factor , growth hormone , and heparin-binding P01133 -like growth factor . To elucidate its role in the beta-cells , the P40763 gene was disrupted in insulin-producing cells in mice ( P40763 -insKO ) , using a cre-mediated gene recombination approach . Unexpectedly , P40763 -insKO mice exhibited an increase in appetite and obesity at 8 weeks of age or older . The mice showed partial leptin resistance , suggesting that expression of the RIP ( rat insulin promoter ) -cre transgene in hypothalamus partially inhibited the appetite-regulating system . Intraperitoneal glucose tolerance tests , performed in non-obese 5-week-old mice , showed that the P40763 -insKO mice were glucose intolerant . Islet perifusion experiments further revealed a deficiency in early-phase insulin secretion . Whereas islet insulin content or islet mass was not affected , expression levels of P11168 , Q09428 , and P15692 were significantly reduced in P40763 -insKO islets . Interestingly , P40763 -insKO mice displayed impaired islet morphology : alpha-cells were frequently seen in central regions of islets . Our present observations demonstrate a unique role of P40763 in maintaining glucose-mediated early-phase insulin secretion and normal islet morphology . Modeling the neurovascular niche : murine strain differences mimic the range of responses to chronic hypoxia in the premature newborn . Preterm birth results in significant cognitive and motor disabilities , but recent evidence suggests that there is variable recovery over time . One possibility that may explain this variable recovery entails variable neurogenic responses in the subventricular zone ( SVZ ) following the period of chronic hypoxia experienced by these neonates . In this report , we have characterized the responses to chronic hypoxia of two mouse strains that represent a wide range of susceptibility to chronic hypoxia . We determined that C57BL/6 pups and neural progenitor cells ( NPCs ) derived from them exhibit a blunted response to hypoxic insult compared with CD-1 pups and NPCs . Specifically , C57BL/6 pups and NPCs exhibited blunted in vivo and in vitro proliferative and increased apoptotic responses to hypoxic insult . Additionally , C57BL/6 NPCs exhibited lower baseline levels and hypoxia-induced levels of selected transcription factors , growth factors , and receptors ( including HIF-1alpha , Q9GZT9 , P23560 , P15692 , P48061 , TrkB , Nrp-1 , P61073 , and NO ) that determine , in part , the responsiveness to chronic hypoxic insult compared with CD-1 pups and NPCs , providing insight into this important and timely problem in perinatology . 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 . 7,12-Dimethylbenz[a]anthracene exposure induces the DNA repair response in neonatal rat ovaries . 7,12-Dimethylbenz[a]anthracene ( DMBA ) destroys ovarian follicles at all stages of development . This study investigated DMBA-induced DNA double strand break ( DSB ) formation with subsequent activation of the ovarian DNA repair response in models of pre-antral or pre-ovulatory follicle loss . Postnatal day ( P01160 ) 4 Fisher 344 ( F344 ) rat ovaries were cultured for 4 days followed by single exposures of vehicle control ( 1 % DB01093 ) or DMBA ( 12.5 nM or 75 nM ) and maintained in culture for 4 or 8 days . Alternately , PND4 F344 rat ovaries were exposed to 1 μM DMBA at the start of culture for 2 days . Total RNA or protein was isolated , followed by qPCR or Western blotting to quantify mRNA or protein level , respectively . γ P16104 and phosphorylated Q13315 were localized and quantified using immunofluorescence staining . DMBA exposure increased caspase 3 and γ P16104 protein . Additionally , DMBA ( 12.5 nM and 1 μM ) increased levels of mRNA encoding Atm , Xrcc6 , Brca1 and Rad51 . In contrast , Parp1 mRNA was decreased on d4 and increased on d8 of DMBA exposure , while P09874 protein increased after 8 days of DMBA exposure . Total Q13315 increased in a concentration-dependent temporal pattern ( 75 nM d4 ; 12.5 nM d8 ) , while pATM was localized in large primary and secondary follicles and increased after 8 days of 75 nM DMBA exposure compared to both control and 12.5 nM DMBA . These findings support that , despite some concentration effects , DMBA induces ovarian DNA damage and that DNA repair mechanisms are induced as a potential mechanism to prevent follicle loss . TIS21(/ P78543 /PC3) accelerates the repair of DNA double strand breaks by enhancing Mre11 methylation and blocking damage signal transfer to the Chk2(T68)-p53(S20) pathway . DNA double strand breaks ( DSBs ) occur more frequently in TIS21(-/-) mouse embryo fibroblasts than that in wild type MEFs ( wt-MEFs ) . Therefore , the role TIS21 plays in the DNA damage response was investigated . Adenoviral transduction of Huh7 tumor cells with the TIS21 gene accelerated the repair of DSBs induced by etoposide treatment as evaluated by clearance of γ P16104 foci and the Comet assay . TIS21 increased methylation of Mre11 and protein arginine methyltransferase 1 ( Q99873 ) activity , leading to Mre11 activation in vitro and in vivo , as determined by immunoprecipitation and radiolabeling analyses . When downstream DNA damage response mediators were evaluated in various human cancer cells lines , TIS21 was found to strongly inhibit Chk2(T68) and p53(S20) phosphorylation by p- Q13315 (S1981) but not p53(S15) . The loss of Chk2 activation after etoposide treatment reduced apoptosis in the cells by downregulating the expression of Q01094 and Bax . These data suggest that TIS21 regulates DSB repair and apoptosis . Expression of TIS21 promoted the repair of DSBs and reduced apoptosis by blocking the damage signal from p- Q13315 (S1981) to Chk2(T68)-p53(S20)via the activation of Mre11 and Q99873 . A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion .

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interacts_with DB01233?

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Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . Regulation of apoptosis signal-regulating kinase 1 degradation by G alpha13 . Apoptosis signal-regulating kinase ( Q99683 ) is a mitogen-activated protein kinase ( MAPK ) that transduces apoptotic signals from a variety of stresses . We have shown previously that alpha subunits of heterotrimeric G12 and Q99941 proteins stimulate Q99683 kinase activity and Q99683 -dependent apoptosis . Here , we report a novel mechanism of G-protein-dependent regulation of Q99683 . We demonstrated that G alpha13 forms a complex with Q99683 in an activation-independent manner . Both N- and C-terminal regulatory domains of Q99683 were essential for the efficient interaction , while its kinase domain was not required . Formation of the G alpha13- Q99683 complex was enhanced by JNK-interacting leucine zipper protein , O60271 . Constitutively activated G alpha13Q226L increased Q99683 expression . Short-term activation of a serotonin Q13639 receptor that is coupled to G alpha13 also increased Q99683 expression . Importantly , prolonged activation of Q13639 receptor in COS-7 cells or prolonged treatment of human umbilical vein endothelial cells with thrombin concomitantly down-regulated both G alpha13 and Q99683 . Data showed that G alpha13Q226L reduced the rate of Q99683 degradation , decreased Q99683 ubiquitination , and reduced association of Q99683 with an E3 ubiquitin ligase Q9UNE7 , previously shown to mediate Q99683 degradation . Our findings indicate that Q99683 expression levels can be regulated by G alpha13 , at least in part via control of Q99683 ubiquitination and degradation . Brominated cyclodipeptides from the marine sponge Geodia barretti as selective 5-HT ligands . The brominated cyclodipeptides barettin ( cyclo[(6-bromo-8-entryptophan)arginine] ) and 8,9-dihydrobarettin ( cyclo[(6-bromotryptophan)arginine] ) isolated from the marine sponge Geodia barretti have previously been shown to inhibit settlement of barnacle larvae in a dose-dependent manner in concentrations ranging from 0.5 to 25 microM . To further establish the molecular target and mode of action of these compounds , we investigated their affinity to human serotonin receptors . The tryptophan residue in the barettins resembles that of endogenous serotonin [ 5-hydroxytryptamine ] . A selection of human serotonin receptors , including representatives from all subfamilies ( 1-7 ) , were transfected into P29320 -293 cells . Barettin selectively interacted with the serotonin receptors 5- Q13049 , P28335 , and Q13639 at concentrations close to that of endogenous serotonin , with the corresponding Ki values being 1.93 , 0.34 , and 1.91 microM , respectively . 8,9-Dihydrobarettin interacted exclusively with the P28335 receptor with a Ki value of 4.63 microM ; it failed to show affinity to 5- Q13049 and Q13639 , indicating that the double bond between the tryptophan and arginine residue plays an important role in the interaction with the receptor proteins . Differential role of the basolateral amygdala 5- Q9H205 and Q13639 serotonin receptors upon ACPA-induced anxiolytic-like behaviors and emotional memory deficit in mice . BACKGROUND AND AIM : The critical role of cannabinoidergic and serotonergic systems of the amygdala in modulation of anxiety-like behaviors and emotional memory has already been demonstrated . The present study aimed to investigate the possible role of the basolateral amygdala ( BLA ) 5- Q9H205 and Q13639 serotonergic systems upon ACPA ( P21554 cannabinoid receptor agonist ) -induced anxiolytic-like behaviors and emotional memory impairment using the elevated plus-maze ( EPM ) test-retest paradigm in male mice . METHOD : bilateral guide-cannulae were implanted to allow intra-BLA microinjection of serotonergic agents . RESULTS : the intraperitoneal injection of ACPA could induce anxiolytic-like behaviors and reduce the emotional memory formation . Intra-BLA injection of M-Chlorophenylbiguanide ( M-Chl , a 5- Q9H205 serotonin receptor agonist ) neither altered the anxiety-like behaviors nor the emotional memory formation by itself , while the higher dose of Y-25130 ( a 5- Q9H205 serotonin receptor antagonist ) reduced the emotional memory formation and locomotor activity but not the anxiety-like behaviors . Furthermore , injection of a higher dose of RS67333 and RS23597 ( as Q13639 serotonin receptor agonist and antagonist , respectively ) did not alter the anxiety-like behaviors , while reduced the emotional memory formation . In addition , the intra-BLA injection of M-Chl but not Y-25130 and RS67333 restored the ACPA-induced anxiolytic-like behaviors and emotional memory deficit , while a higher dose of RS67333 decreased the locomotor activity . Moreover , the intra-BLA microinjection of RS23597 could restore the ACPA-induced anxiolytic-like behaviors but not the emotional memory deficit . CONCLUSION : based on our findings , ACPA seems to induce its anxiolytic-like behaviors and emotional memory formation deficits via activation and deactivation of the BLA Q13639 and 5- Q9H205 serotonin receptors . Human enteroendocrine cell responses to infection with Chlamydia trachomatis : a microarray study . BACKGROUND : Enteroendocrine cells ( EEC ) are highly specialized cells producing signalling molecules vital to the normal functions of the gut . Recently , we showed altered protein distribution in Chlamydia infected EEC in vitro . The aim of this study was to perform a microarray analysis of the response pattern of EEC from both large and small bowel to infection in vitro , using Chlamydia trachomatis infection as a model . METHODS : TWO HUMAN EEC LINES : LCC-18 , derived from a neuroendocrine colonic tumour , and CNDT-2 , derived from a small intestinal carcinoid , were infected using cultured C. trachomatis serovar LGV II strain 434 ( ATCC VR-902B ) . DB01053 was used to induce persistent infection . We used microarray analysis ( Affymetrix GeneChip® ) for studying changes in gene expression at different stages of infection . RESULTS : Twenty-four hours after active and persistent infection , 66 and 411 genes in LCC-18 and 68 and 170 genes in CNDT-2 cells , respectively showed mean expression ratios > 2-fold compared to non-infected cells . These genes encoded factors regulating apoptosis , cell differentiation , transcription regulation , cytokine activity , amine biosynthesis and vesicular transport . We found significant differences in gene transcription levels between persistently infected and non-infected cells in 10 genes coding for different solute carrier transporters ( O00585 ) and in 5 genes related to endocrine function ( Q9H0R8 , GRIP1 , P14416 , O00445 and O43581 ) . CONCLUSIONS : Infected EEC cells exhibit cell-type specific patterns related to vesicular transport , secretion and neurotransmitters . EEC play a pivotal role in regulation of gut motility and an impairment of enteroendocrine function can contribute to motility disorders . Transforming growth factor alpha-induced expression of type 1 plasminogen activator inhibitor in astrocytes rescues neurons from excitotoxicity . Although transforming growth factor ( TGF ) -alpha , a member of the epidermal growth factor ( P01133 ) family , has been shown to protect neurons against excitotoxic and ischemic brain injuries , its mechanism of action remains unknown . In the present study , we used in vitro models of apoptotic or necrotic paradigms demonstrating that TGF-alpha rescues neurons from N-methyl-D-aspartate ( DB01221 ) -induced excitotoxic death , with the obligatory presence of astrocytes . Because neuronal tissue-type plasminogen activator ( t-PA ) release was shown to potentiate DB01221 -induced excitotoxicity , we observed that TGF-alpha treatment reduced DB01221 -induced increase of t-PA activity in mixed cultures of neurons and astrocytes . In addition , we showed that although TGF-alpha induces activation of the extracellular signal-regulated kinases ( ERKs ) in astrocytes , it failed to activate Q8NFH3 / Q8TCB0 in neurons . Finally , we showed that TGF-alpha , by an P29323 -dependent mechanism , stimulates the astrocytic expression of P05121 , a t-PA inhibitor , which mediates the neuroprotective activity of TGF-alpha against DB01221 -mediated excitotoxic neuronal death . Taken together , we indicate that TGF-alpha rescues neurons from DB01221 -induced excitotoxicity in mixed cultures through inhibition of t-PA activity , involving P05121 overexpression by an P29323 -dependent pathway in astrocytes . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Effects of metoclopramide and tropisetron on aldosterone secretion possibly due to agonism and antagonism at the Q13639 receptor . OBJECTIVE : Part of the prokinetic activity of metoclopramide can possibly be ascribed to agonist activity at Q13639 receptors . The 5- Q9H205 antagonist tropisetron is thought to act as an antagonist at Q13639 receptors . In the present study aldosterone secretion in response to the administration of these two drugs was explored to examine the role of the Q13639 receptor in aldosterone secretion . METHODS : Following a single-blind , random design , ten normal male volunteers received one of the following regimens on three occasions , with at least 2-week intervals : metoclopramide 10 mg i.v. ; tropisetron 5 mg by slow i.v.i. , or ; tropisetron by slow i.v.i. , followed by 10 mg metoclopramide i.v. RESULTS : In response to metoclopramide alone the mean plasma aldosterone level rose significantly to 149 % of basal level and remained significantly elevated for the next 20 min . With tropisetron alone , there was a significant 37.8 % drop at 60 min and the aldosterone levels remained low for the duration of the experiment . DB01233 reversed the decline mediated by tropisetron significantly at 30 and 90 min . DB04630 levels after the latter regimen also did not differ significantly from baseline at any time period . CONCLUSION : These results would suggest the existence of a tonic stimulatory influence of 5-HT via Q13639 receptors on aldosterone secretion , which could be augmented by metoclopramide and blocked by tropisetron . However , the effect of tropisetron per se should be interpreted with caution given the lack of a saline group . P04150 -mediated regulation of P14780 gene expression in human ovarian surface epithelial cells . OBJECTIVE : To obtain proof-of-concept that locally produced anti-inflammatory steroids suppress ovulation-associated extracellular matrix proteases in human ovarian surface epithelial ( OSE ) cells . DESIGN : Primary OSE cell cultures treated with interleukin-1alpha ( IL-1alpha ) ( 500 pg/mL ) as proxy for inflammation , with/without anti-inflammatory steroid ( cortisol or progesterone [ P ] , 0.01-1.0 microM ) . SETTING : Academic medical center . PATIENT(S) : Sixteen premenopausal women ( 29-46 years ) undergoing surgery for nonmalignant gynecological conditions . MAIN OUTCOME MEASURE(S) : Semiquantitative extracellular matrix protease gene expression profiling with verification by real-time quantitative reverse transcription polymerase chain reaction ( qRT-PCR ) and gelatinase zymography . RESULT(S) : Treatment with IL-1alpha stimulated messenger RNA ( mRNA ) expression of several ovulation-associated matrix metalloproteinase genes by OSE cell cultures , including gelatinase B ( P14780 ) but not gelatinase A ( P08253 ) . The IL-1alpha-stimulated P14780 mRNA production was suppressed by cortisol but not P. DB00741 but not P also dose-dependently suppressed IL-1alpha-stimulated P14780 gelatinase activity and this effect was blocked by the glucocorticoid receptor antagonist DB00834 . CONCLUSION(S) : In human OSE cells , stimulation of P14780 gene expression and proteolytic activity by IL-1alpha is suppressed by anti-inflammatory cortisol through a glucocorticoid receptor-mediated mechanism . Because IL-1alpha also generates cortisol formation in OSE by stimulating cortisone reductase activity , these results support a role for intracrine cortisol in minimizing proteolytic damage to the OSE at ovulation . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . DB01233 does not increase gastric muscle contractility in newborn rats . Feeding intolerance resulting from delayed gastric emptying is common in premature neonates . DB01233 ( MCP ) , the most frequently used prokinetic drug in neonates , enhances gastric muscle contractility through inhibition of dopamine receptors . Although its therapeutic benefit is established in adults , limited data are available to support its clinical use in infants . Hypothesizing that developmentally dependent differences are present , we comparatively evaluated the effect of MCP on fundus muscle contractility in newborn , juvenile , and adult rats . The muscle strips were either contracted with electrical field stimulation ( O43281 ) to induce cholinergic nerve-mediated acetylcholine release or carbachol , a cholinergic agonist acting directly on the muscarinic receptor . Although in adult rats MCP increased O43281 -induced contraction by 294 ± 122 % of control ( P < 0.01 ) , no significant effect was observed in newborn fundic muscle . MCP had no effect on the magnitude of the carbachol-induced and/or bethanechol-induced gastric muscle contraction at any age . In response to dopamine , an 80.7 ± 5.3 % relaxation of adult fundic muscle was observed , compared with only a 8.4 ± 8.7 % response in newborn tissue ( P < 0.01 ) . P14416 expression was scant in neonates and significantly increased in adult gastric tissue ( P < 0.01 ) . In conclusion , the lack of MCP effect on the newborn fundic muscle contraction potential relates to developmental differences in dopamine D2 receptor expression . To the extent that these novel data can be extrapolated to neonates , the therapeutic value of MCP as a prokinetic agent early in life requires further evaluation . Dopamine-related genes and their relationships to monoamine metabolites in P04141 . Monoamine metabolite ( MM ) levels in lumbar cerebrospinal fluid ( P04141 ) are extensively used as indirect estimates of monoamine turnover in the brain . In this study we investigated genotypes for DNA polymorphisms in the D2 ( P14416 ) , D3 ( P35462 ) , and D4 ( P21917 ) dopamine receptor and tyrosine hydroxylase ( TH ) genes and their relationships to P04141 MM in healthy volunteers ( n = 66 ) . Concentrations of homovanillic acid ( HVA ) , 3-methoxy-4-hydroxyphenylglycol ( MHPG ) , and 5-hydroxyindoleacetic acid ( 5-HIAA ) were corrected for back length , a confounding variable . Corrected MM levels were not related to age , gender , height , weight heredity , season or atmospheric pressure at sampling . Individuals with specific P14416 and TH allele and genotype configurations significantly differed in HVA and MHPG concentrations . P35462 homo- and heterozygotic genotypes had significantly different P04141 5-HIAA levels . P21917 genotypes were not related to MM concentrations . The results suggest that specific P14416 , P35462 , and TH genotypes participate in the regulation of monoamine turnover in the central nervous system . Accordingly monoamine receptors and synthesizing enzyme genotypes appear to be variance factors influencing MM concentrations in P04141 . The relationships found in this study support MM concentrations as markers for monoamine transmission in the human brain . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . The relationship between gastric motility and nausea : gastric prokinetic agents as treatments . Nausea is one of a cluster of symptoms described subjectively by patients with delayed gastric emptying . The mechanisms and treatments are unclear ( anti-emetic drugs are not fully effective against nausea ) . Can nausea be relieved by stimulating gastric emptying ? Physostigmine ( together with atropine ) has been shown experimentally to stimulate gastric motility , relieve nausea and restore normal gastric motility . Is this mimicked by gastric prokinetic drugs ? The answer is complicated by mixed pharmacology . DB01233 increases gastric motility by activating myenteric Q13639 receptors but also directly inhibits vomiting via D2 and 5- Q9H205 receptor antagonism ; relationships between increased gastric motility and relief from nausea are therefore unclear . Similarly , the D2 receptor antagonist domperidone has direct anti-emetic activity . Nevertheless , more selective Q13639 and motilin receptor agonists ( erythromycin , directly stimulating gastric motility ) inhibit vomiting in animals ; low doses of erythromycin can also relieve symptoms in patients with gastroparesis . Ghrelin stimulates gastric motility and appetite mostly via vagus-dependent pathways , and inhibits vomiting in animals . To date , ghrelin receptor activation has failed to consistently improve gastric emptying or symptoms in patients with gastroparesis . We conclude that nausea can be relieved by gastric prokinetic drugs , but more clinical studies are needed using drugs with selective activity . Other mechanisms ( e.g. ghrelin , vagal and central pathways , influencing a mechanistic continuum between appetite and nausea ) also require exploration . These and other issues will be further explored in a forthcoming special issue of the European Journal of Pharmacology , which focusses on mechanisms of nausea and vomiting . DB02546 shows preferential cytotoxicity in mutant p53 cancer cells by destabilizing mutant p53 through inhibition of the Q9UBN7 -Hsp90 chaperone axis . Mutant p53 ( mutp53 ) cancers are surprisingly dependent on their hyperstable mutp53 protein for survival , identifying mutp53 as a potentially significant clinical target . However , exploration of effective small molecule therapies targeting mutp53 has barely begun . Mutp53 hyperstabilization , a hallmark of p53 mutation , is cancer cell-specific and due to massive upregulation of the HSP90 chaperone machinery during malignant transformation . We recently showed that stable complex formation between HSP90 and its mutp53 client inhibits E3 ligases Q00987 and Q9UNE7 , causing mutp53 stabilization . Histone deacetylase ( HDAC ) inhibitors ( HDACi ) are a new class of promising anti-cancer drugs , hyperacetylating histone and non-histone targets . Currently , suberoylanilide hydroxamic acid ( DB02546 ) is the only FDA-approved HDACi . We show that DB02546 exhibits preferential cytotoxicity for mutant , rather than wild-type and null p53 human cancer cells . Loss/gain-of-function experiments revealed that although able to exert multiple cellular effects , DB02546 's cytotoxicity is caused to a significant degree by its ability to strongly destabilize mutp53 at the level of protein degradation . The underlying mechanism is DB02546 's inhibition of Q9UBN7 , an essential positive regulator of HSP90 . This releases mutp53 and enables its Q00987 - and Q9UNE7 -mediated degradation . DB02546 also strongly chemosensitizes mutp53 cancer cells for chemotherapy due to its ability to degrade mutp53 . This identifies a novel action of DB02546 with the prospect of DB02546 becoming a centerpiece in mutp53-specific anticancer strategies . Human and mouse trace amine-associated receptor 1 have distinct pharmacology towards endogenous monoamines and imidazoline receptor ligands . TAARs ( trace amine-associated receptors ) are G-protein-coupled receptors that respond to low abundance , endogenous amines such as tyramine and tryptamine , and represent potential targets for neuropsychiatric diseases . However , some members of this receptor subfamily either have no ligand identified or remain difficult to express and characterize using recombinant systems . In the present paper we report the successful expression of human and mouse Q96RJ0 , and the characterization of their responses to various natural and synthetic agonists . In P29320 ( human embryonic kidney ) -293/CRE-bla cells , mouse Q96RJ0 showed a robust response to trace amines as measured using either a DB02527 assay or a beta-lactamase reporter assay , whereas human Q96RJ0 showed a weaker , but still measurable , response . When certain fragments of human Q96RJ0 were replaced with the corresponding regions of mouse Q96RJ0 , the chimaeric receptor showed a much stronger response in DB02527 production . Examination of a series of agonists on these receptors revealed that the human and the chimaeric receptor are almost identical in pharmacology , but distinct from the mouse receptor . We also screened small libraries of pharmacologically active agents on Q96RJ0 and identified a series of synthetic agonists , some of which are also ligands of the enigmatic imidazoline receptor . The findings of the present study not only shed light on the pharmacological species difference of Q96RJ0 , but also raise new possibilities about the mechanism of some of the imidazoline-related agents . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . Identification of novel genes regulated in the developing human ventral mesencephalon . In the human embryo , from approximately 6 weeks gestational age ( GA ) , dopaminergic ( DA ) neurons can be found in the ventral mesencephalon ( VM ) . More specifically , the post-mitotic neurons are located in the ventral part of the tegmentum ( VT ) , whereas no mature DA neurons are found in the neighboring dorsal part . We used Affymetrix HG-U133 GeneChip technology to compare genome-wide expression profiles of ventral and dorsal tegmentum from 8 weeks GA human embryos , in order to identify genes involved in specification , differentiation , and survival of mesencephalic DA ( mDA ) neurons . Known mDA marker genes including P00352 , Q01959 , Q05940 , TH , P05937 , P43354 , P55317 , P48051 , O75364 , P07949 , and P14416 topped the list of 96 genes from HG-U133A with higher expression in VT , validating the experimental set-up . In addition , 28 probes from HG-U133B were identified whereof most are annotated to UniGene clusters with no gene associated or to genes of unknown function . Of these , the fifteen most regulated transcripts , representing changes down to 56 % could be verified by quantitative real-time PCR ( Q-PCR ) on a developmental series of subdissected human embryonic and fetal brain material , resulting in not only a regional but also a temporal expression profile . This revealed a distinct DA-associated profile for in particular a putative transcription factor ( FLJ45455 ) and the uncharacterized transmembrane proteins Q9ULS5 and Q96EP9 . The data presented here may help to device cell replacement and regenerative therapies for Parkinson 's disease ( PD ) . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . 5-HT₄ receptor stimulation leads to soluble AβPPα production through P14780 upregulation . Serotonin 4 ( Q13639 ) receptor signaling does not only have the physiological function of improving cognition , but might also be helpful in the therapy of Alzheimer 's disease ( AD ) through regulation of the production of soluble amyloid-β protein precursor alpha ( sAβPPα ) . To analyze the relationship between Q13639 receptor signaling and sAβPPα production , we stably transfected H4 cells with AβPP and Q13639 receptor ( H4/AβPP/ Q13639 cells ) . We found that 24-h incubation with the Q13639 receptor agonist RS-67333 upregulates matrix metalloproteinase-9 ( P14780 ) . Furthermore , P14780 overexpression enhanced sAβPPα levels , whereas knockdown with P14780 siRNA decreased sAβPPα levels . When RS-67333 was injected for 10 days in Tg2576 mice , a model of amyloid-β peptide ( Aβ ) deposition , there was an increase in hippocampal levels of sAβPPα , C-terminal fragment α , and P14780 , as well as a decrease in hippocampal senile plaque number and levels of the 40 amino acid peptide , Aβ40 . Taken all together , these experiments demonstrate that Q13639 receptor stimulation induces expression of P14780 which cleaves AβPP through α-secretase-like activity , leading to an increase of sAβPPα levels and a reduction of Aβ load . Polyamines and membrane transporters . In recent years , our understanding of the importance of membrane transporters ( MTs ) in the disposition of and response to drugs has increased significantly . MTs are proteins that regulate the transport of endogenous molecules and xenobiotics across the cell membrane . In mammals , two super-families have been identified : DB00171 -binding cassette ( DB01048 ) and solute carrier ( O00585 ) transporters . There is evidence that MTs might mediate polyamines ( PA ) transport . PA are ubiquitous polycations which are found in all living cells . In mammalian cells , three major PA are synthesised : putrescine , spermidine and spermine ; whilst the decarboxylated arginine ( agmatine ) is not produced by mammals but is synthesised by plants and bacteria . In addition , research in the PA field suggests that PA are transported into cells via a specific transporter , the polyamine transport system(s) ( Q03393 ) . Although the Q03393 has not been fully defined , there is evidence that some of the known MTs might be involved in PA transport . In this mini review , eight O00585 transporters will be reviewed and their potential to mediate PA transport in human cells discussed . These transporters are O15245 , O15244 , O75751 , Q96FL8 , P30825 , P08195 , SLC12A8A , and Q86VW1 . Preliminary data from our laboratory have revealed that O15245 might be involved in the PA uptake ; in addition to one member of ABC superfamily ( P08183 protein ) might also mediate the efflux of polyamine like molecules . DB01233 stimulates catecholamine- and granin-derived peptide secretion from pheochromocytoma cells through activation of serotonin type 4 ( Q13639 ) receptors . The gastroprokinetic agent metoclopramide is known to stimulate catecholamine secretion from pheochromocytomas . The aim of the study was to investigate the mechanism of action of metoclopramide and expression of serotonin type 4 ( 5-HT(4) ) receptors in pheochromocytoma tissues . Tissue explants , obtained from 18 pheochromocytomas including the tumor removed from a 46-year-old female patient who experienced life-threatening hypertension crisis after metoclopramide administration and 17 additional pheochromocytomas ( 9 benign and 8 malignant ) were studied . Cultured pheochromocytoma cells derived from the patient who previously received metoclopramide were incubated with metoclopramide and various 5-HT(4) receptor ligands . In addition , total mRNAs were extracted from all the 18 tumors . Catecholamine- and granin-derived peptide concentrations were measured in pheochromocytoma cell incubation medium by HPLC and radioimmunological assays . In addition , expression of 5-HT(4) receptor mRNAs in the 18 pheochromocytomas was investigated by the use of reverse transcriptase-PCR . RESULTS : DB01233 and the 5-HT(4) receptor agonist cisapride were found to activate catecholamine- and granin-derived peptide secretions by cultured tumor cells . DB01233 - and cisapride-evoked catecholamine- and granin-derived peptide productions were inhibited by the 5-HT(4) receptor antagonist GR 113808 . 5-HT(4) receptor mRNAs were detected in the patient 's tumor and the series of 17 additional pheochromocytomas . This study shows that pheochromocytomas express functional 5-HT(4) receptors that are responsible for the stimulatory action of metoclopramide on catecholamine- and granin-derived peptide secretion . All 5-HT(4) receptor agonists must therefore be contraindicated in patients with proven or suspected pheochromocytoma .

MH_train_9

MH_train_9

MH_train_9

interacts_with DB00277?

multiple_choice

Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Antenatal maternally-administered phosphodiesterase type 5 inhibitors normalize P29474 expression in the fetal lamb model of congenital diaphragmatic hernia . PURPOSE : Pulmonary hypertension ( pHTN ) , a main determinant of survival in congenital diaphragmatic hernia ( Q8NE62 ) , results from in utero vascular remodeling . Phosphodiesterase type 5 ( O76074 ) inhibitors have never been used antenatally to treat pHTN . The purpose of this study is to determine if antenatal O76074 inhibitors can prevent pHTN in the fetal lamb model of Q8NE62 . METHODS : Q8NE62 was created in pregnant ewes . Postoperatively , pregnant ewes received oral placebo or tadalafil , a O76074 inhibitor , until delivery . Near term gestation , lambs underwent resuscitations , and lung tissue was snap frozen for protein analysis . RESULTS : Mean cGMP levels were 0.53±0.11 in placebo-treated fetal lambs and 1.73±0.21 in tadalafil-treated fetal lambs ( p=0.002 ) . Normalized expression of P29474 was 82 % ±12 % in Normal-Placebo , 61 % ±5 % in Q8NE62 -Placebo , 116 % ±6 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . Normalized expression of β-sGC was 105 % ±15 % in Normal-Placebo , 82 % ±3 % in Q8NE62 -Placebo , 158 % ±16 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . P29474 and β-sGC were significantly decreased in Q8NE62 ( p=0.0007 and 0.01 for P29474 and β-sGC , respectively ) , and tadalafil significantly increased P29474 expression ( p=0.0002 ) . CONCLUSIONS : O76074 inhibitors can cross the placental barrier . β-sGC and P29474 are downregulated in fetal lambs with Q8NE62 . Antenatal O76074 inhibitors normalize P29474 and may prevent in utero vascular remodeling in Q8NE62 . Pressure overload-induced cardiac hypertrophy response requires janus kinase 2-histone deacetylase 2 signaling . Pressure overload induces cardiac hypertrophy through activation of O60674 ( Jak2 ) , however , the underlying mechanisms remain largely unknown . In the current study , we tested whether histone deacetylase 2 ( Q92769 ) was involved in the process . We found that angiotensin II ( Ang-II ) -induced re-expression of fetal genes ( Atrial natriuretic peptide ( P01160 ) and brain natriuretic peptide ( DB04899 ) ) in cultured cardiomyocytes was prevented by the Jak2 inhibitor AG-490 and Q92769 inhibitor Trichostatin-A ( P32119 ) , or by Jak2/ Q92769 siRNA knockdown . On the other hand , myocardial cells with Jak2 or Q92769 over-expression were hyper-sensitive to Ang-II . In vivo , pressure overload by transverse aorta binding ( AB ) induced a significant cardiac hypertrophic response as well as re-expression of P01160 and DB04899 in mice heart , which were markedly reduced by AG-490 and P32119 . Significantly , AG-490 , the Jak2 inhibitor , largely suppressed pressure overload-/Ang-II-induced Q92769 nuclear exportation in vivo and in vitro . Meanwhile , P32119 or Q92769 siRNA knockdown reduced Ang-II-induced P01160 / DB04899 expression in Jak2 over-expressed H9c2 cardiomyocytes . Together , these results suggest that Q92769 might be a downstream effector of Jak2 to mediate cardiac hypertrophic response by pressure overload or Ang-II . [ The significance of inflammatory markers in sputum of asthmatic and chronic obstructive pulmonary diseases patients before and after glucocorticoid treatment ] . OBJECTIVE : To study the change of cytokines and eosinophil cationic protein ( P12724 ) level in the sputum before and after glucocorticoid ( GC ) inhalation treatment so as to comprehend their effect on asthmatic and chronic obstructive pulmonary diseases ( P48444 ) patients . METHODS : A method to induce sputum with inhaled hypertonic saline was used . The level of interleukin ( IL ) -5 , P10145 and P12724 was measured with enzyme-linked immunosorbent assay method . RESULTS : The concentration of P12724 decreased from ( 500.3 +/- 49.6 ) microg/L to ( 59.8 +/- 10.9 ) microg/L , the percentage of eosinophils ( Eos ) dropped from ( 11.6 +/- 1.7 ) x 10(-2) to ( 4.1 +/- 0.7 ) x 10(-2) and there is significant difference in the concentration of P05113 in the group of asthmatic patients after GC treatment . However , the concentration of P05113 in the P48444 patients did not show significant change after the same therapy . CONCLUSION : Respiratory tract inflammation in asthma is related to Eos activation and increase in P12724 and P05113 excretion , while respiratory tract inflammation in P48444 is related to neutrophil increase . These changes can be considered as the indicator of airway inflammation in asthma or P48444 . Through regulating the quantity and function of the inflammatory cells and inhibiting the formation of cytokines to control the asthmatic airway inflammation , GC inhalation treatment will have better effect in treating asthmatic patients than P48444 patients . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation . Expression of cytosolic retinoid-binding protein genes in human skin biopsies and cultured keratinocytes and fibroblasts . Using reverse transcription coupled to polymerase chain reaction we have studied the mRNA expression of serum retinol-binding protein and cytosolic receptors for retinol and retinoic acid in skin biopsies , and in cultured epidermal keratinocytes and dermal fibroblasts . Transcripts for cellular retinol-binding protein ( P09455 ) I and cellular retinoic-acid-binding protein ( CRABP ) I were found in normal skin , keratinocytes , and fibroblasts . CRABP II transcripts were detected in skin and keratinocytes . A decreased mRNA expression of CRABP I and an increased mRNA expression of CRABP II were found in lesional psoriatic skin compared with uninvolved skin . mRNA transcripts for serum retinol-binding protein ( s- P02753 ) were detected in all tissues and cells . The biological importance of s- P02753 expression in keratinocytes and fibroblasts is not known , but hypothetically this protein may be involved in the intracellular shuttling of retinol and retinoic acid , or in the retransportation of cellular retinoids into the extracellular space . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Branched N-glycans and their implications for cell adhesion , signaling and clinical applications for cancer biomarkers and in therapeutics . Branched N-glycans are produced by a series of glycosyltransferases including N-acetylglucosaminyltransferases and fucosyltransferases and their corresponding genes . Glycans on specific glycoproteins , which are attached via the action of glycosyltransferases , play key roles in cell adhesion and signaling . Examples of this are adhesion molecules or signaling molecules such as integrin and P12830 , as well as membrane receptors such as the P01133 and TGFβ receptors . These molecules also play pivotal roles in the underlying mechanism of a variety of disease such as cancer metastasis , diabetes , and chronic obstructive pulmonary disease ( P48444 ) . Alterations in the structures of branched N-glycans are also hall marks and are useful for cancer biomarkers and therapeutics against cancer . This mini-review describes some of our recent studies on a functional glycomics approach to the study of branched N-glycans produced by N-acetylglucosaminyltransferases III , IV , V and IX ( Vb ) ( GnT-III , GnT-IV , V and IX ( Vb ) ) and fucosyltransferase 8 ( Fut8 ) and their patho-physiological significance , with emphasis on the importance of a systems glycobiology approach as a future perspective for glycobiology . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . How corticosteroids control inflammation : Quintiles Prize Lecture 2005 . Corticosteroids are the most effective anti-inflammatory therapy for many chronic inflammatory diseases , such as asthma but are relatively ineffective in other diseases such as chronic obstructive pulmonary disease ( P48444 ) . Chronic inflammation is characterised by the increased expression of multiple inflammatory genes that are regulated by proinflammatory transcription factors , such as nuclear factor-kappaB and activator protein-1 , that bind to and activate coactivator molecules , which then acetylate core histones to switch on gene transcription . Corticosteroids suppress the multiple inflammatory genes that are activated in chronic inflammatory diseases , such as asthma , mainly by reversing histone acetylation of activated inflammatory genes through binding of liganded glucocorticoid receptors ( GR ) to coactivators and recruitment of histone deacetylase-2 ( Q92769 ) to the activated transcription complex . At higher concentrations of corticosteroids GR homodimers also interact with DNA recognition sites to active transcription of anti-inflammatory genes and to inhibit transcription of several genes linked to corticosteroid side effects . In patients with P48444 and severe asthma and in asthmatic patients who smoke Q92769 is markedly reduced in activity and expression as a result of oxidative/nitrative stress so that inflammation becomes resistant to the anti-inflammatory actions of corticosteroids . DB00277 , by activating HDAC , may reverse this corticosteroid resistance . This research may lead to the development of novel anti-inflammatory approaches to manage severe inflammatory diseases . Inhibition of inducible nitric oxide synthase in respiratory diseases . DB00435 ( NO ) is a key physiological mediator and disturbed regulation of NO release is associated with the pathophysiology of almost all inflammatory diseases . A multitude of inhibitors of NOSs ( nitric oxide synthases ) have been developed , initially with low or even no selectivity against the constitutively expressed NOS isoforms , P29474 ( endothelial NOS ) and P29475 ( neuronal NOS ) . In the meanwhile these efforts yielded potent and highly selective P35228 ( inducible NOS ) inhibitors . Moreover , P35228 inhibitors have been shown to exert beneficial anti-inflammatory effects in a wide variety of acute and chronic animal models of inflammation . In the present mini-review , we summarize some of our current knowledge of inhibitors of the P35228 isoenzyme , their biochemical properties and efficacy in animal models of pulmonary diseases and in human disease itself . Moreover , the potential benefit of P35228 inhibition in animal models of P48444 ( chronic obstructive pulmonary disease ) , such as cigarette smoke-induced pulmonary inflammation , has not been explicitly studied so far . In this context , we demonstrated recently that both a semi-selective P35228 inhibitor { L-NIL [ N6-(1-iminoethyl)-L-lysine hydrochloride ] } and highly selective P35228 inhibitors ( GW274150 and BYK402750 ) potently diminished inflammation in a cigarette smoke mouse model mimicking certain aspects of human P48444 . Therefore , despite the disappointing results from recent asthma trials , P35228 inhibition could still be of therapeutic utility in P48444 , a concept which needs to be challenged and validated in human disease . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Histone deacetylase-2 and airway disease . The increased expression of inflammatory genes in inflammatory lung diseases is regulated by acetylation of core histones , whereas histone deacetylase-2 ( Q92769 ) suppresses inflammatory gene expression . Corticosteroids suppress inflammatory genes in asthma by inhibiting histone acetyltransferase and in particular by recruiting Q92769 to the nuclear factor-kappaB-activated inflammatory gene complex . This involves deacetylation of the acetylated glucocorticoid receptor . In P48444 , severe asthma and asthmatics who smoke , Q92769 is reduced , thus preventing corticosteroids from suppressing inflammation . The reduction in Q92769 appears to be secondary to increased oxidative and nitrative stress in the lungs . Antioxidants and inhibitors of nitric oxide synthesis may therefore restore corticosteroid sensitivity in P48444 , but this can also be achieved by low concentrations of theophylline and curcumin , which act as HDAC activators . DB00277 is a direct inhibitor of oxidant-activated phosphoinositide-3-kinase-delta , which is involved in inactivation of Q92769 . In the future selective O00329 inhibitors and more direct activators of Q92769 may be used to treat corticosteroid-resistant inflammatory diseases of the lung , including P48444 , severe asthma and asthma in smokers . Genetic alterations and oncogenic pathways associated with breast cancer subtypes . Breast cancers can be divided into subtypes with important implications for prognosis and treatment . We set out to characterize the genetic alterations observed in different breast cancer subtypes and to identify specific candidate genes and pathways associated with subtype biology . mRNA expression levels of estrogen receptor , progesterone receptor , and P04626 were shown to predict marker status determined by immunohistochemistry and to be effective at assigning samples to subtypes . P04626 (+) cancers were shown to have the greatest frequency of high-level amplification ( independent of the P04626 amplicon itself ) , but triple-negative cancers had the highest overall frequencies of copy gain . Triple-negative cancers also were shown to have more frequent loss of phosphatase and tensin homologue and mutation of P06400 , which may contribute to genomic instability . We identified and validated seven regions of copy number alteration associated with different subtypes , and used integrative bioinformatics analysis to identify candidate oncogenes and tumor suppressors , including P04626 , Q14451 , O95251 , O15297 , P24385 , Q92769 , P55317 , and P20936 . We tested the candidate oncogene O95251 and showed that it enhances the anchorage-independent growth of breast cancer cells . The genome-wide and region-specific differences between subtypes suggest the differential activation of oncogenic pathways . Loss of corepressor O15055 under hypoxia up-regulates OCT1-mediated EMT gene expression and enhances tumor malignancy . The circadian clock gene Period2 ( O15055 ) has been suggested to be a tumor suppressor . However , detailed mechanistic evidence has not been provided to support this hypothesis . We found that loss of O15055 enhanced invasion and activated expression of epithelial-mesenchymal transition ( EMT ) genes including Q15672 , O43623 , and SNAIL . This finding was corroborated by clinical observation that O15055 down-regulation was associated with poor prognosis in breast cancer patients . We further demonstrated that O15055 served as a transcriptional corepressor , which recruited polycomb proteins Q15910 and Q15022 as well as Q92769 to octamer transcription factor 1 ( OCT1 ) ( P14859 ) binding sites of the Q15672 and O43623 promoters to repress expression of these EMT genes . Hypoxia , a condition commonly observed in tumors , caused O15055 degradation and disrupted the O15055 repressor complex , leading to activation of EMT gene expression . This result was further supported by clinical data showing a significant negative correlation between hypoxia and O15055 . Thus , our findings clearly demonstrate the tumor suppression function of O15055 and elucidate a pathway by which hypoxia promotes EMT via degradation of O15055 . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . Evidence of an Epigenetic Modification in Cell-cycle Arrest Caused by the Use of Ultra-highly-diluted Gonolobus Condurango Extract . OBJECTIVES : Whether the ultra-highly-diluted remedies used in homeopathy can effectively bring about modulations of gene expressions through acetylation/deacetylation of histones has not been explored . Therefore , in this study , we pointedly checked if the homeopathically-diluted anti-cancer remedy Condurango 30C ( ethanolic extract of Gonolobus condurango diluted 10(-60) times ) was capable of arresting the cell cycles in cervical cancer cells HeLa by triggering an epigenetic modification through modulation of the activity of the key enzyme histone deacetylase 2 vis-a-vis the succussed alcohol ( placebo ) control . METHODS : We checked the activity of different signal proteins ( like P38936 (WAF) , p53 , Akt , P40763 ) related to deacetylation , cell growth and differentiation by western blotting and analyzed cell-cycle arrest , if any , by fluorescence activated cell sorting . After viability assays had been performed with Condurango 30C and with a placebo , the activities of histone de-acetylase ( HDAC ) enzymes 1 and 2 were measured colorimetrically . RESULTS : While Condurango 30C induced cytotoxicity in HeLa cells in vitro and reduced Q92769 activity quite strikingly , it apparently did not alter the Q13547 enzyme ; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity . Data on P38936 (WAF) , p53 , Akt , and P40763 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and P55008 -phase cell-cycle arrest when Condurango 30C was used at a 2 % dose . CONCLUSION : Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells , which the placebo was unable to do . [ The nutritional status and respiratory function of patients diagnosed with P48444 ] . Given that patients affected with chronic obstructive pulmonary disease show a progressive weight loss , and the great socioeconomic repercussions of these diseases due to their high incidence in the general population , we started this study with the objective of analyzing the possible connection between the nutritional state and the ventilatory function of a group of patients from our midst , who did not have continuous oxygen therapy at home . We studied a total of 43 patients who had been diagnosed with P48444 ( excluding those with a BMI < 32 ) , evaluating anthropometric , biochemical and pulmonary function parameters . Among the obtained results , it should be noted that 84 % of the patients were normally nourished and only 16 % were undernourished . In the lung function analysis , we found a pattern of air flow obstruction . We found a significant correlation between the nutritional state and the type of P48444 ( p < 0.01 ) , with the emphysematous patients being more undernourished than those suffering from bronchial disease . We also found a significant correlation between the types of P48444 and the levels of prealbumin , P02753 and albumin , and a positive correlation between the evolution time of the disease and the levels of albumin , PO2 and FEV1 . With the obtained results , we do not consider it necessary to establish a nutritional support protocol in ambulatory patients suffering from P48444 , whose conditions are similar to those of the patients in our study group . P01308 resistance is not exhibited by advanced chronic obstructive pulmonary disease patients . We have previously reported increased blood glucose concentrations and skeletal muscle glycogen depletion in severe P48444 patients with chronic respiratory failure . In order to see if insulin resistance exists in severe P48444 , we investigated nine patients with advanced P48444 with chronic hypoxaemia and seven healthy control subjects of similar age , using the euglycaemic hyperinsulinaemic glucose clamp technique . We could not demonstrate a subnormal intravenous glucose requirement in response to insulin when maintaining euglycaemia in the P48444 patients with chronic hypoxaemia . This indicates that the net metabolism of glucose in P48444 patients with chronic hypoxaemia is not resistant to insulin . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone .

MH_train_10

MH_train_10

MH_train_10

interacts_with DB01367?

multiple_choice

Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . Free energy force field ( FEFF ) 3D-QSAR analysis of a set of Plasmodium falciparum dihydrofolate reductase inhibitors . Free energy force field ( FEFF ) 3D-QSAR analysis was used to construct ligand-receptor binding models for a set of 18 structurally diverse antifolates including pyrimethamine , cycloguanil , methotrexate , aminopterin and trimethoprim , and 13 pyrrolo[2,3-d]pyrimidines . The molecular target ( ' receptor ' ) used was a 3D-homology model of a specific mutant type of Plasmodium falciparum ( Pf ) dihydrofolate reductase ( P00374 ) . The dependent variable of the 3D-QSAR models is the IC50 inhibition constant for the specific mutant type of PfDHFR . The independent variables of the 3D-QSAR models ( the descriptors ) are scaled energy terms of a modified first-generation AMBER force field combined with a hydration shell aqueous solvation model and a collection of 2D-QSAR descriptors often used in QSAR studies . Multiple temperature molecular dynamics simulation ( P43034 ) and the genetic function approximation ( GFA ) were employed using partial least square ( PLS ) and multidimensional linear regressions as the fitting functions to develop FEFF 3D-QSAR models for the binding process . The significant FEFF energy terms in the best 3D-QSAR models include energy contributions of the direct ligand-receptor interaction . Some changes in conformational energy terms of the ligand due to binding to the enzyme are also found to be important descriptors . The FEFF 3D-QSAR models indicate some structural features perhaps relevant to the mechanism of resistance of the PfDHFR to current antimalarials . The FEFF 3D-QSAR models are also compared to receptor-independent ( RI ) 4D-QSAR models developed in an earlier study and subsequently refined using recently developed generalized alignment rules . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Patient age and biological aggressiveness of endometrial carcinoma . BACKGROUND : Advanced age is associated with a significantly worse prognosis of endometrial carcinoma patients . The aim of this study was to test whether age is a poor-risk factor in endometrial carcinoma because tumors arising in older patients are biologically different from those diagnosed in patients of an earlier age . MATERIALS AND METHODS : DB03843 -fixed , paraffin-embedded samples from 136 previously untreated patients with endometrial carcinoma were studied by means of immunohistochemistry . The expression of molecular markers associated with hormone responsiveness ( estrogen and progesterone receptors ) , proliferation ( Ki67 , C-ERB-B2 , p53 ) , invasiveness ( P12830 ) and apoptosis ( P10415 and p53 ) was analyzed . The obtained expression levels , together with all available clinical and pathological features were tested for correlations with the patients age and survival . RESULTS : Advanced patient age showed a direct correlation with tumor stage ( r=0.29 , p=0.0008 ) and mutant p53 expression ( r=0.25 , p=0.004 ) , and an inverse correlation with P12830 expression ( r=-0.28 , p=0.001 ) . Patient age above the 25th percentile ( 57 years ) of the age distribution was significantly associated with a worse prognosis ( p=0.018 ) . CONCLUSION : It appears that with advancing age , endometrial carcinoma exhibits a more aggressive tumor phenotype , characterized by mutant p53 expression and down-regulation of P12830 expression , and that this , in its turn , results in tumors being diagnosed at a more advanced stage in older patients . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Reactive oxygen species , DNA damage , and error-prone repair : a model for genomic instability with progression in myeloid leukemia ? Myelodysplastic syndromes ( P43034 ) comprise a heterogeneous group of disorders characterized by ineffective hematopoiesis , with an increased propensity to develop acute myelogenous leukemia ( AML ) . The molecular basis for P43034 progression is unknown , but a key element in P43034 disease progression is loss of chromosomal material ( genomic instability ) . Using our two-step mouse model for myeloid leukemic disease progression involving overexpression of human mutant P01111 and P10415 genes , we show that there is a stepwise increase in the frequency of DNA damage leading to an increased frequency of error-prone repair of double-strand breaks ( DSB ) by nonhomologous end-joining . There is a concomitant increase in reactive oxygen species ( ROS ) in these transgenic mice with disease progression . Importantly , P63000 , an essential component of the ROS-producing NADPH oxidase , is downstream of DB01367 , and we show that ROS production in P01111 / P10415 mice is in part dependent on P63000 activity . DNA damage and error-prone repair can be decreased or reversed in vivo by N-acetyl cysteine antioxidant treatment . Our data link gene abnormalities to constitutive DNA damage and increased DSB repair errors in vivo and provide a mechanism for an increase in the error rate of DNA repair with P43034 disease progression . These data suggest treatment strategies that target DB01367 / P31749 pathways and ROS production in human P43034 /AML . Role of adiponectin in delayed embryonic development of the short-nosed fruit bat , Cynopterus sphinx . The aim of this study was to evaluate the role of adiponectin in the delayed embryonic development of Cynopterus sphinx . Q15848 receptor ( Q96A54 ) abundance was first observed to be lower during the delayed versus non-delayed periods of utero-embryonic unit development . The effects of adiponectin treatment on embryonic development were then evaluated during the period of delayed development . Exogenous treatment increased the in vivo rate of embryonic development , as indicated by an increase in weight , Q96A54 levels in the utero-embryonic unit , and histological changes in embryonic development . Treatment with adiponectin during embryonic diapause showed a significant increase in circulating progesterone and estradiol concentrations , and in production of their receptors in the utero-embryonic unit . The adiponectin-induced increase in estradiol synthesis was correlated with increased cell survival ( P10415 protein levels ) and cell proliferation ( P12004 protein levels ) in the utero-embryonic unit , suggesting an indirect effect of adiponectin via estradiol synthesis by the ovary . An in vitro study further confirmed the in vivo findings that adiponectin treatment increases P12004 levels together with increased uptake of glucose by increasing the abundance of glucose transporter 8 ( GLUT8 ) in the utero-embryonic unit . The in vitro study also revealed that adiponectin , together with estradiol but not alone , significantly increased Q96A54 protein levels . Thus , adiponectin works in concert with estradiol to increase glucose transport to the utero-embryonic unit and promote cell proliferation , which together accelerate embryonic development . Induction of apoptosis by a dominant negative H- DB01367 mutant ( 116Y ) in K562 cells . Recent extensive work on apoptosis has begun to reveal its molecular mechanisms . Several genes that regulate apoptosis have been identified . Among them , the P10415 gene is considered to be an important gene that inhibits apoptosis . However , there must be other genes , yet to be identified , which suppress apoptosis . It has been suggested that the activation of DB01367 function by P11274 - P00519 fusion protein in chronic myelogenous leukemia may be an important mechanism in the P11274 - P00519 mediated transformation . Therefore , in this study we have investigated whether the suppression of endogenous H- DB01367 function inhibits the P11274 - P00519 mediated transforming activity in a K562 human chronic myelogenous leukemia cell line . The induced expression of a dominant negative v-H- DB01367 mutant ( 116Y ) in K562 cells has resulted in cell death . The morphological characteristics and the detection of fragmented DNA by gel electrophoresis in the dead cells have revealed that this cell death is apoptosis . These results directly indicate that the DB01367 gene as well as the P10415 gene has an ability to suppress apoptosis . The BH3 mimetic DB05764 synergizes with the Q02750 /2 inhibitor selumetinib/AZD6244 to promote O43521 -dependent tumour cell death and inhibit acquired resistance . Tumour cells typically exhibit a G(1) cell cycle arrest in response to the Q02750 /2 [ mitogen-activated protein kinase/ P29323 ( extracellular-signal-regulated kinase ) kinase 1/2 ] inhibitor selumetinib , but do not die , and thus they acquire resistance . In the present study we examined the effect of combining selumetinib with the BH3 [ P10415 ( B-cell lymphoma 2 ) homology domain 3 ] -mimetic P10415 inhibitor DB05764 . Although either drug alone caused little tumour cell death , the two agents combined to cause substantial caspase-dependent cell death and inhibit long-term clonogenic survival of colorectal cancer and melanoma cell lines with P15056 (V600E) or DB01367 mutations . This cell death absolutely required Q07812 ( P10415 -associated X protein ) and was inhibited by RNAi ( RNA interference ) -mediated knockdown of O43521 ( P10415 -interacting mediator of cell death ) in the P15056 (V600E)-positive COLO205 cell line . When colorectal cancer cell lines were treated with selumetinib plus DB05764 we observed a striking reduction in the incidence of cells emerging with acquired resistance to selumetinib . Similar results were observed when we combined DB05764 with the P15056 (V600E)-selective inhibitor PLX4720 , but only in cells expressing P15056 (V600E) . Finally , cancer cells in which acquired resistance to selumetinib arises through P15056 (V600E) amplification remained sensitive to DB05764 , whereas selumetinib-resistant HCT116 cells ( P01116 (G13D) amplification ) were cross-resistant to DB05764 . Thus the combination of a P10415 inhibitor and an P27361 /2 pathway inhibitor is synthetic lethal in P27361 /2-addicted tumour cells , delays the onset of acquired resistance and in some cases overcomes acquired resistance to selumetinib . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Estrogen induced changes in Akt-dependent activation of endothelial nitric oxide synthase and vasodilation . OBJECTIVES : Acute administration of estrogen results in vasodilation and increased nitric oxide ( NO ) production . We examined the hypothesis that this is due to activation of Akt/ P31749 which subsequently increases P29474 activity . METHODS AND RESULTS : Treatment of bovine microvascular and human umbilical endothelial cells ( HUVEC ) with 17-beta-estradiol ( E2 ) ( 10(-9) to 10(-5)M ) increased phosphorylation of Akt within 1 min and this was followed by phosphorylation of P29474 . These effects were blocked by wortmannin , a PI(3)K inhibitor and the upstream activator of Akt . The estrogen receptor antagonist , DB00947 , inhibited P29474 phosphorylation . E2 increased calcium dependent NOS activity and nitrite production and this was inhibited by wortmannin and DB00947 . E2 increased the vasodilatory response of aortic rings to acetylcholine and wortmannin blocked the effect . E2 ( 10(-9)M ) dilated cerebral microvascular vessels under conditions of no flow , constant flow and increasing flow and this was blocked by wortmannin . Tamoxifen , a partial estrogen receptor antagonist , also dilated the microvessels . CONCLUSIONS : : E2 increases NO production through an Akt/ P31749 dependent pathway . This is associated with increased sensitivity to endothelial dependent dilation . In cerebral microvessels , E2 and tamoxifen produce significant dilation at low concentrations with and without acetylcholine induced stimulation of endothelial vasodilation . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . Transformed 3T3 cells have reduced levels and altered subcellular distribution of the major PKC substrate protein P29966 . The P29966 ( myristylated alanine-rich C-kinase substrate ) protein is an abundant calmodulin-binding protein that is a major and specific endogenous substrate of protein kinase C ( PKC ) . Stimulation of cells with phorbol esters or other activators of PKC has been shown previously to result in rapid phosphorylation of P29966 proteins and redistribution of these myristylated C-kinase substrates from membrane to cytosol . Here we show that NIH3T3 murine fibroblasts transformed by P38936 -HA-C- DB01367 or pp60-V- P12931 oncoproteins have markedly reduced levels of p68- P29966 and that most of the remaining P29966 protein is found in the cytosol . 3T3 cells containing a nontransforming oncoprotein Q9Y3Q3 - P10415 , in contrast , exhibited normal levels and distribution of p68- P29966 . When taken together with recent evidence that P29966 proteins are involved in regulating organization of the membrane cytoskeleton , our findings suggest that oncoprotein-mediated alterations in P29966 protein levels and subcellular distribution may contribute to the development or maintenance of the transformed phenotype . Murine models of chronic lymphocytic leukaemia : role of microRNA-16 in the New Zealand Black mouse model . Mouse models are valuable tools in the study of human chronic lymphocytic leukaemia ( CLL ) . The New Zealand Black ( NZB ) strain is a naturally occurring model of late-onset CLL characterized by B-cell hyperproliferation and autoimmunity early in life , followed by progression to CLL . Other genetically engineered models of CLL that have been developed include ( NZB x NZW ) F1 mice engineered to express P05113 , mice expressing human P56279 , and mice overexpressing both P10415 and a tumour necrosis factor receptor-associated factor . The applicability to human CLL varies with each model , suggesting that CLL is a multifactorial disease . Our work with the de novo NZB model has revealed many similarities to the human situation , particularly familial CLL . In NZB , the malignant clones express P06127 , zap-70 , and have chromosomal instability and germline Ig sequence . We also identified a point mutation in the 3'-flanking sequence of Mirn16-1 , which resulted in decreased levels of the microRNA , miR-16 in lymphoid tissue . Exogenous restoration of miR-16 to an NZB malignant B-1 cell line resulted in cell cycle alterations , suggesting that the altered expression of Mirn15a/16-1 is an important molecular lesion in CLL . Future studies utilizing the NZB mouse could ascertain the role of environmental triggers , such as low dose radiation and organic chemicals in the augmentation of a pre-existing propensity to develop CLL . Cooperative Hedgehog- P00533 signaling . It has been known for many years that cooperative interactions between oncogenes ( e.g. DB01367 , MYC , P10415 ) can fuel cancer growth ( 1-5 ) , but the restricted druggability of many of those interacting cancer genes has hampered translation of combined targeting to medical cancer therapy . The identification and characterization of cooperative cancer signaling pathways amenable to medical therapy is therefore a crucial step towards the establishment of efficient targeted combination treatments urgently needed to improve cancer therapy . Here we review recent findings of our group and colleagues on the molecular mechanisms of cooperative Hedgehog/ P08151 and Epidermal Growth Factor Receptor ( P00533 ) signaling , two clinically relevant oncogenic pathways involved in the development of many human malignancies . We also discuss the possible implications of these findings for the design of a therapeutic regimen relying on combined targeting of key effectors of both pathways . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Differential role of two P11473 coactivators , Q15648 and Q9Y6Q9 , in keratinocyte proliferation and differentiation . Cell programs such as proliferation and differentiation involve the selective activation and repression of gene expression . The vitamin D receptor ( P11473 ) , through 1,25(OH)(2)D(3) , controls the proliferation and differentiation of keratinocytes . Previously , we have identified two P11473 binding coactivator complexes . In proliferating keratinocytes P11473 bound preferentially to the DRIP complex , whereas in differentiated keratinocytes the P12931 complex was preferred . We proposed that different coactivators are required for sequential gene regulation in the transition from proliferation to differentiation . Here we examined the roles of Q15648 and Q9Y6Q9 in this transition . Silencing of Q15648 and P11473 caused hyperproliferation of keratinocytes , demonstrated by increased XTT and BrdU incorporation . Q9Y6Q9 silencing , on the other hand , did not have an effect on proliferation . In contrast , Q9Y6Q9 as well as Q15648 and P11473 silencing blocked keratinocyte differentiation as shown by decreased expression of keratin 1 and filaggrin . These results are consistent with the differential localization of Q15648 and Q9Y6Q9 in skin . These results indicate that Q15648 is required for keratinocyte proliferation . Both Q15648 and Q9Y6Q9 are required for the keratinocyte differentiation . These results support the concept that the selective use of coactivators by P11473 underlies the selective regulation of gene expression in keratinocyte proliferation and differentiation . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Polymorphisms in DNA repair and apoptosis-related genes and clinical outcomes of patients with non-small cell lung cancer treated with first-line paclitaxel-cisplatin chemotherapy . This study was conducted to analyze a comprehensive panel of single nucleotide polymorphisms ( SNPs ) in genes in DNA repair and apoptosis pathways and determine the relationship between polymorphisms and treatment outcomes of patients with non-small cell lung cancer ( NSCLC ) treated with first-line paclitaxel-cisplatin chemotherapy . Three hundred eighty two patients with NSCLC were enrolled . Seventy-four SNPs in 48 genes ( 42 SNPs in 27 DNA repair pathway genes and 32 SNPs in 21 apoptotic pathway genes ) were genotyped and their associations with chemotherapy response and overall survival ( OS ) were analyzed . Among SNPs in DNA repair genes , P38398 rs799917 was significantly associated with both chemotherapy response and OS . P18887 rs25487 exhibited a significant association with chemotherapy response and P18074 rs1052555 with OS . Four SNPs in apoptotic genes ( P20333 rs1061624 , P10415 rs2279115 , O15392 rs9904341 , and Q14790 rs3769818 ) were significantly associated with OS , but not with response to chemotherapy . When the six SNPs which were associated with OS in individual analysis were combined , OS decreased as the number of bad genotypes increased ( P(trend) = 2 × 10(-6) ) . Patients with 3 , and 4-6 bad genotypes had significantly worse OS compared with those carrying 0-2 bad genotypes ( adjusted hazard ratio [ aHR ] = 1.54 , 95 % CI = 1.14-2.08 , P = 0.005 ; aHR = 2.10 , 95 % CI = 1.55-2.85 , P = 2 × 10(-6) , respectively ) . In conclusion , these findings suggest that the six SNPs identified , particularly their combined genotypes , could be used as biomarkers predicting chemotherapy response and survival of NSCLC patients treated with first-line paclitaxel-cisplatin chemotherapy . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Class I histone deacetylase activity is required for proliferation of renal epithelial cells . The process of renal regeneration after acute kidney injury is thought to recapitulate renal development , and proliferation of renal proximal tubular cells ( RPTCs ) is a critical step in the regenerative response . Recent studies indicate that class I histone deacetylases ( HDACs ) are required for embryonic kidney gene expression , growth , and differentiation . The role and underlying mechanisms of class I HDAC activation in RPTC proliferation , however , remain unclear . In this study , we used cultured RPTCs to examine this issue since four class I HDAC isoforms ( 1 , 2 , 3 , and 8 ) are abundantly expressed in this cell type . Blocking class I HDAC activity with a highly selective inhibitor , MS-275 , induced global histone H3 hyperacetylation , reduced RPTC proliferation , and diminished expression of cyclin D1 and proliferating cell nuclear antigen . Silencing Q13547 , 3 , or 8 with small interfering RNA resulted in similar biological effects . Activation of epidermal growth factor receptor ( P00533 ) and signal transducers and activators of transcription 3 ( P40763 ) was required for RPTC proliferation , and P40763 functioned downstream of P00533 . Treatment with MS-275 or knockdown of Q13547 , 3 , or 8 suppressed P00533 expression and phosphorylation , and silencing Q13547 and 3 also reduced P40763 phosphorylation . However , Q92769 downregulation did not affect RPTC proliferation and phosphorylation of P00533 and P40763 . Collectively , these data reveal a critical role of class I HDACs in mediating proliferation of renal epithelial cells through activation of the P00533 / P40763 signaling pathway . Personalized medicine and pharmacogenetic biomarkers : progress in molecular oncology testing . In the field of oncology , clinical molecular diagnostics and biomarker discoveries are constantly advancing as the intricate molecular mechanisms that transform a normal cell into an aberrant state in concert with the dysregulation of alternative complementary pathways are increasingly understood . Progress in biomarker technology , coupled with the companion clinical diagnostic laboratory tests , continue to advance this field , where individualized and customized treatment appropriate for each individual patient define the standard of care . Here , we discuss the current commonly used predictive pharmacogenetic biomarkers in clinical oncology molecular testing : P15056 V600E for vemurafenib in melanoma ; Q9HC35 - Q9UM73 for crizotinib and P00533 for erlotinib and gefitinib in non-small-cell lung cancer ; P01116 against the use of cetuximab and panitumumab in colorectal cancer ; P04626 ( P04626 /neu ) for trastuzumab in breast cancer ; P11274 - P00519 for tyrosine kinase inhibitors in chronic myeloid leukemia ; and P29590 /RARα for all-trans-retinoic acid and arsenic trioxide treatment for acute promyelocytic leukemia . Molecular pathways : the basis for rational combination using MEK inhibitors in P01116 -mutant cancers . Mutations in DB01367 oncogenes are frequently observed in human cancers , and the mutations result in activation of the DB01367 -RAF-MEK- P29323 pathway , leading to cell proliferation and survival . The pathway is , therefore , a potent therapeutic target in the DB01367 -mutant cancers . MEK inhibitors can specifically block the pathway and are one of the key types of drugs for the treatment of the DB01367 -mutant cancers . As DB01367 proteins activate other downstream signaling proteins in addition to the DB01367 -RAF-MEK- P29323 pathway , combination therapeutic approaches with MEK inhibitors are also being evaluated . Moreover , MEK inhibitors can arrest cancer cells in P55008 phase and repress prosurvival Bcl2 family proteins such as Q07820 and P10415 /BCLXL , and increase expression of Bim , a proapoptotic BH3-only family protein . This mechanism may explain the efficacy of the combination of MEK inhibitors with cytotoxic agents or other targeted inhibitors . A better understanding of the pathway will help us with development of rational combinations for the treatment of the DB01367 -mutant cancers .

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interacts_with DB09079?

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Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . DB09079 , a triple angiokinase inhibitor , enhances cytotoxic therapy response in pancreatic cancer . Angiogenesis remains a sensible target for pancreatic ductal adenocarcinoma ( PDAC ) therapy . P15692 , PDGF , FGF and their receptors are expressed at high levels and correlate with poor prognosis in human PDAC . DB09079 is a triple angiokinase inhibitor that targets P17948 /2/3 , P11362 /2/3 and PDGFRα/β signaling . We investigated the antitumor activity of nintedanib alone or in combination with the cytotoxic agent gemcitabine in experimental PDAC . DB09079 inhibited proliferation of cells from multiple lineages found in PDAC , with gemcitabine enhancing inhibitory effects . DB09079 blocked PI3K/MAPK activity and induced apoptosis in vitro and in vivo . In a heterotopic model , net local tumor growth compared to controls ( 100 % ) was 60.8 ± 10.5 % in the gemcitabine group , -2.1 ± 9.9 % after nintedanib therapy and -12.4 ± 16 % after gemcitabine plus nintedanib therapy . Effects of therapy on intratumoral proliferation , microvessel density and apoptosis corresponded with tumor growth inhibition data . In a PDAC survival model , median animal survival after gemcitabine , nintedanib and gemcitabine plus nintedanib was 25 , 31 and 38 days , respectively , compared to 16 days in controls . The strong antitumor activity of nintedanib in experimental PDAC supports the potential of nintedanib-controlled mechanisms as targets for improved clinical PDAC therapy . Sustained vascular endothelial growth factor delivery enhances angiogenesis and perfusion in ischemic hind limb . PURPOSE : We hypothesized that sustained delivery of vascular endothelial growth factor ( P15692 ) using a polymer [ 85:15 poly(lactide-co-glycolide) ( P00747 ) ] would enhance angiogenesis and improve perfusion of ischemic tissue . METHODS : C57BL/6J mice ( n = 20/group ) underwent unilateral hind limb ischemia surgery and were randomized to groups of no scaffold implantation ( 0-Implant ) , unloaded scaffold implantation ( Empty- P00747 ) , or implantation of scaffolds incorporating 3 microg of VEGF165 ( P00747 - P15692 ) . Endpoints included laser Doppler perfusion imaging ( LDPI , ischemic/nonischemic limb , % ) , local vessel counts , immunohistochemistry for CD31 , and alpha-smooth muscle actin . In vitro release kinetics of P15692 from P00747 was also measured . RESULTS : P00747 - P15692 resulted in improved lower extremity perfusion vs. controls as measured by LDPI % at 7 , 14 , 21 , and 28 days ( p < 0.05 ) . P00747 - P15692 was associated with significantly greater percentage of vessels staining for CD31 and alpha-smooth muscle actin compared to the Empty- P00747 or 0-Implant ( p < 0.05 for both ) . CONCLUSIONS : The P00747 - P15692 scaffolds resulted in sustained P15692 delivery , improved tissue perfusion , greater capillary density , and more mature vasculature compared to the controls . The sustained-release P00747 polymer vehicle is a promising delivery system for therapeutic neovascularization applications . [ 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 . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . DB00819 inhibits aquaporin-1 expression and colon cancer xenograft tumor growth . BACKGROUND/AIMS : To study the effects of water channel protein inhibitor acetazolamide on xenograft tumor growth of colon cancer in nude mice . METHODOLOGY : Setting up human colon cancer model in nude mice , mice were randomly divided into two groups as experimental group and control group . DB00819 was given at a volume of 0.1mL per mice ( 40mg/kg/d , ig ) in experimental group , while the same volume of sterile saline was given in control group ( ig ) . After 21 days , protein and m-RNA levels of P29972 in tumor tissues from two groups were detected respectively by Western blot and RT-PCR to evaluate the treatment effects . P29972 , P15692 and P28906 expression was detected by immunohistochemistry , simultaneously . RESULTS : DB00819 ( 40mg/kg/d , ig ) significantly inhibited the xenograft tumor growth of colon cancer in nude mice . The inhibition rate was 88.28 % . In comparison with the control group , P29972 protein and mRNA level were significantly reduced in the experimental group ( p < 0.01 ) . P29972 , P15692 and P28906 expression in experimental group were positively correlated between each other ( p < 0.01 ) . CONCLUSIONS : DB00819 can suppress the xenograft tumor growth by inhibiting the expression of P29972 . Modeling the neurovascular niche : murine strain differences mimic the range of responses to chronic hypoxia in the premature newborn . Preterm birth results in significant cognitive and motor disabilities , but recent evidence suggests that there is variable recovery over time . One possibility that may explain this variable recovery entails variable neurogenic responses in the subventricular zone ( SVZ ) following the period of chronic hypoxia experienced by these neonates . In this report , we have characterized the responses to chronic hypoxia of two mouse strains that represent a wide range of susceptibility to chronic hypoxia . We determined that C57BL/6 pups and neural progenitor cells ( NPCs ) derived from them exhibit a blunted response to hypoxic insult compared with CD-1 pups and NPCs . Specifically , C57BL/6 pups and NPCs exhibited blunted in vivo and in vitro proliferative and increased apoptotic responses to hypoxic insult . Additionally , C57BL/6 NPCs exhibited lower baseline levels and hypoxia-induced levels of selected transcription factors , growth factors , and receptors ( including HIF-1alpha , Q9GZT9 , P23560 , P15692 , P48061 , TrkB , Nrp-1 , P61073 , and NO ) that determine , in part , the responsiveness to chronic hypoxic insult compared with CD-1 pups and NPCs , providing insight into this important and timely problem in perinatology . Lymphagenesis correlates with expression of vascular endothelial growth factor-C in colorectal cancer . Lymphagenesis in gastrointestinal tumors is not well described . To clarify its presence and regulation , we assessed the microlymphatic count ( MLC ) in colorectal cancer patients . Lymphatic vessels were evaluated by enzyme-histochemistry for 5'-nucleotidase ( 5'-NA ) . Since vascular endothelial growth factor ( P15692 ) -C is reportedly associated with lymphagenesis , the expression of P49767 protein was immunohistochemically assessed by the catalyzed signal amplification ( Q13216 ) method . MLC of peritumoral lesions was significantly higher than that of non-cancer and intratumoral lesions ( p < 0.01 ) ; it increased where P49767 was highly expressed ( p < 0.01 ) and increased with the depth of invasion in peritumoral lesions . These results indicate significant findings at peritumoral lesion : that lymphagenesis may be elicited by tumor spread ; that P49767 expression is associated with lymphagenesis and is a potent factor stimulating lymphagenesis . A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins . A surface plasmon resonance-based solution affinity assay is described for measuring the K(d) of binding of heparin/heparan sulfate-binding proteins with a variety of ligands . The assay involves the passage of a pre-equilibrated solution of protein and ligand over a sensor chip onto which heparin has been immobilised . DB01109 sensor chips prepared by four different methods , including biotin-streptavidin affinity capture and direct covalent attachment to the chip surface , were successfully used in the assay and gave similar K(d) values . The assay is applicable to a wide variety of heparin/HS-binding proteins of diverse structure and function ( e.g. , P05230 , P09038 , P15692 , P10145 , P80075 , P01008 , P02776 ) and to ligands of varying molecular weight and degree of sulfation ( e.g. , heparin , DB05808 , sucrose octasulfate , naphthalene trisulfonate ) and is thus well suited for the rapid screening of ligands in drug discovery applications . 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 . Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries . BACKGROUND : Atherosclerosis affects aorta , coronary , carotid , and iliac arteries most frequently than any other body vessel . There may be common molecular pathways sustaining this process . Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis . RESULTS : We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression . We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis . Caveolae and JAK/ P35610 pathways , and P06702 / P05109 interacting proteins are certainly involved in the development of vascular disease . We found that the system of caveolae is directly connected with genes that respond to hormone receptors , and indirectly with the apoptosis pathway . Cytokines , chemokines and growth factors released in the blood flux were investigated in parallel . High levels of RANTES , IL-1ra , MIP-1 alpha , MIP-1 beta , P60568 , P05112 , P05113 , P05231 , P13232 , Q16552 , DB00102 , P15692 and P01579 were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels . CONCLUSION : The pattern of cytokine and P06702 / P05109 up-regulation characterizes atherosclerosis as a proinflammatory disorder . Activation of the JAK/ P35610 pathway is confirmed by the up-regulation of P05231 , P42224 , Q00978 and Q13651 genes in coronary and carotid plaques . The functional network constructed in our research is an evidence of the central role of P35610 protein and the caveolae system to contribute to preserve the plaque . Moreover , Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP .

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Emerging therapeutic targets in bladder cancer . Treatment of muscle invasive urothelial bladder carcinoma ( BCa ) remains a major challenge . Comprehensive genomic profiling of tumors and identification of driver mutations may reveal new therapeutic targets . This manuscript discusses relevant molecular drivers of the malignant phenotype and agents with therapeutic potential in BCa . Small molecule pan-FGFR inhibitors have shown encouraging efficacy and safety results especially among patients with activating FGFR mutations or translocations . P42345 inhibitors for patients with Q92574 mutations and concomitant targeting of PI3K and MEK represent strategies to block PI3K/AKT/ P42345 pathway . Encouraging preclinical results with ado-trastuzumab emtansine ( DB05773 ) exemplifies a new potential treatment for P04626 -positive BCa along with innovative bispecific antibodies . Inhibitors of cell cycle regulators ( aurora kinase , polo-like kinase 1 , and cyclin-dependent kinase 4 ) are being investigated in combination with chemotherapy . Early results of clinical studies with anti- P16410 and anti- Q9NZQ7 are propelling immune modulating drugs to the forefront of emerging treatments for BCa . Collectively , these novel therapeutic targets and treatment strategies hold promise to improve the outcome of patients afflicted with this malignancy . DB05773 -associated telangiectasias in metastatic breast cancer : a case series . Treatment of P04626 -positive metastatic breast cancer with ado-trastuzumab emtansine ( DB05773 ) , a novel antibody-drug conjugate , has resulted in both improved progression-free and overall survival . Recognition and treatment of diverse adverse events related to DB05773 is critical for safety and tolerability . The most frequent adverse events with DB05773 include fatigue , diarrhea , anemia , elevated transaminases , and mild-to-moderate hemorrhagic events , which are thought to be related to induced thrombocytopenia . Here , we present five case series of cutaneous and mucosal telangiectasias , definitely related to DB05773 . The development of telangiectasias represents a newly recognized adverse effect of DB05773 . We provide description and timing of the telangiectasias and review the mechanisms that may explain the formation of these vascular lesions in association with DB05773 . Further , we describe associated bleeding events and propose that induced telangiectasias could represent an additional cause of DB05773 -associated hemorrhage . DB05773 ( Kadcyla ) for P04626 -positive metastatic breast cancer . Advanced P04626 -positive gastric cancer : current and future targeted therapies . The prognostic value of human epidermal growth factor receptor 2 ( P04626 ) in gastric cancer is controversial . Consensus guidelines have standardized the testing of P04626 status in gastric cancer . Overexpression of this receptor occurs in approximately 20 % of gastric and gastro-esophageal junction adenocarcinomas , predominantly those of the intestinal type . Recently , trastuzumab has emerged as the first targeted drug to improve overall survival when combined with chemotherapy in advanced P04626 -positive gastric cancer . Primary and secondary resistance to trastuzumab has become a major problem and new strategies to overcome this resistance are needed . A high percentage of advanced P04626 -positive gastric cancer patients who progress on trastuzumab therapy are candidates for second-line therapy . New families of targeted drugs , including tyrosine kinase inhibitors ( TKIs ) such as lapatinib and PF-00299804 , mammalian target of rapamycin ( P42345 ) pathway inhibitors such as everolimus , heat-shock protein 90 ( HSP90 ) inhibitors such as AUY922 , HER dimerization inhibitors such as pertuzumab , and antibody-chemotherapy conjugates such as trastuzumab-emtansine ( DB05773 ) , could offer alternative second-line treatments when trastuzumab-based first-line therapy fails . Pharmacological approach to acute pancreatitis . The aim of the present review is to summarize the current knowledge regarding pharmacological prevention and treatment of acute pancreatitis ( AP ) based on experimental animal models and clinical trials . Somatostatin ( SS ) and octreotide inhibit the exocrine production of pancreatic enzymes and may be useful as prophylaxis against post endoscopic retrograde cholangiopancreatography pancreatitis ( PEP ) . The protease inhibitor gabexate mesilate ( GM ) is used routinely as treatment to AP in some countries , but randomized clinical trials and a meta-analysis do not support this practice . Nitroglycerin ( P04626 ) is a nitrogen oxide ( NO ) donor , which relaxes the sphincter of Oddi . Studies show conflicting results when applied prior to ERCP and a large multicenter randomized study is warranted . Steroids administered as prophylaxis against PEP has been validated without effect in several randomized trials . The non-steroidal anti-inflammatory drugs ( NSAID ) indomethacin and diclofenac have in randomized studies showed potential as prophylaxis against PEP . Interleukin 10 ( P22301 ) is a cytokine with anti-inflammatory properties but two trials testing P22301 as prophylaxis to PEP have returned conflicting results . Antibodies against tumor necrosis factor-alpha ( P01375 ) have a potential as rescue therapy but no clinical trials are currently being conducted . The antibiotics beta-lactams and quinolones reduce mortality when necrosis is present in pancreas and may also reduce incidence of infected necrosis . Evidence based pharmacological treatment of AP is limited and studies on the effect of potent anti-inflammatory drugs are warranted . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . The effect of different linkers on target cell catabolism and pharmacokinetics/pharmacodynamics of trastuzumab maytansinoid conjugates . DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate consisting of the anti- P04626 antibody trastuzumab linked via a nonreducible thioether linker to the maytansinoid antitubulin agent DM1 . DB05773 has shown favorable safety and efficacy in patients with P04626 -positive metastatic breast cancer . In previous animal studies , DB05773 exhibited better pharmacokinetics ( PK ) and slightly more efficacy than several disulfide-linked versions . The efficacy findings are unique , as other disulfide-linked antibody-drug conjugates ( ADC ) have shown greater efficacy than thioether-linked designs . To explore this further , the in vitro and in vivo activity , PK , and target cell activation of DB05773 and the disulfide-linked T- Q8TCT9 -DM1 were examined . Both ADCs showed high in vitro potency , with DB05773 displaying greater potency in two of four breast cancer cell lines . In vitro target cell processing of DB05773 and T- Q8TCT9 -DM1 produced lysine-N(ε)-MCC-DM1 , and lysine-N(ε)- Q8TCT9 -DM1 and DM1 , respectively ; in vivo studies confirmed these results . The in vitro processing rates for the two conjugate to their respective catabolites were similar . In vivo , the potencies of the conjugates were similar , and T- Q8TCT9 -DM1 had a faster plasma clearance than DB05773 . Slower DB05773 clearance translated to higher overall tumor concentrations ( conjugate plus catabolites ) , but unexpectedly , similar levels of tumor catabolite . These results indicate that , although the ADC linker can have clear impact on the PK and the chemical nature of the catabolites formed , both linkers seem to offer the same payload delivery to the tumor . DB05773 for P04626 -positive metastatic breast cancer . A population pharmacokinetic/pharmacodynamic model of thrombocytopenia characterizing the effect of trastuzumab emtansine ( DB05773 ) on platelet counts in patients with P04626 -positive metastatic breast cancer . PURPOSE : DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate in the development for the treatment of human epidermal growth factor receptor 2-positive cancers . Thrombocytopenia ( DB01520 ) is the dose-limiting toxicity of DB05773 . A semimechanistic population pharmacokinetic/pharmacodynamic ( PK/PD ) model was developed to characterize the effect of DB05773 on patient platelet counts . METHODS : A PK/PD model with transit compartments that mimic platelet development and circulation was fit to concentration-platelet-time course data from two DB05773 single-agent studies ( TDM3569g ; N = 52 and TDM4258g ; N = 112 ) . NONMEM(®) 7 software was used for model development . Data from a separate phase II study ( TDM4374g ; N = 110 ) were used for model evaluation . Patient baseline characteristics were evaluated as covariates of model PD parameters . RESULTS : The model described the platelet data well and predicted the incidence of grade ≥3 DB01520 . The model predicted that with DB05773 3.6 mg/kg given every 3 weeks ( q3w ) , the lowest platelet nadir would occur after the first dose . Also predicted was a patient subgroup ( 46 % ) having variable degrees of downward drifting platelet-time profiles , which were predicted to stabilize by the eighth treatment cycle to platelet counts above grade 3 DB01520 . Baseline characteristics were not significant covariates of PD parameters in the model . CONCLUSIONS : This semimechanistic PK/PD model accurately captures the cycle 1 platelet nadir , the downward drift noted in some patient platelet-time profiles , and the ~8 % incidence of grade ≥3 DB01520 with DB05773 3.6 mg/kg q3w . This model supports DB05773 3.6 mg/kg q3w as a well-tolerated dose with minimal dose delays or reductions for DB01520 . The potential for trastuzumab emtansine in human epidermal growth factor receptor 2 positive metastatic breast cancer : latest evidence and ongoing studies . The treatment of breast cancer that is driven by amplification and overexpression of human epidermal growth factor receptor 2 ( P04626 ) has been drastically improved by the development of P04626 -targeted therapies including trastuzumab and lapatinib . While outcomes for patients diagnosed with P04626 -positive breast cancer have been greatly impacted by these therapies , treatment resistance is common and toxicity to standard regimens remains a therapeutic challenge . DB00072 emtansine ( DB05773 ) is a novel antibody drug conjugate that consists of the P04626 -targeted monoclonal antibody , trastuzumab , joined via a stable linker to a derivative of maytansine , a highly potent cytotoxic chemotherapy . While other antibody drug conjugates have been developed clinically , this is the first in its class that maintains the antitumor properties of the P04626 -targeted antibody , trastuzumab , and also avoids release of the chemotherapy until the molecule is taken up inside the P04626 -overexpressing cancer cell . Several phase I studies have shown DB05773 is safe , tolerable and has activity in trastuzumab- and lapatinib-pretreated breast cancer . Moreover , phase II studies are now being reported that confirm its safety and clinical efficacy in both the frontline and heavily pretreated settings . Preliminary data from phase II studies evaluating its use in combination with other cytotoxics have also been reported and several large phase III trials are underway to evaluate its use in the P04626 -positive metastatic breast cancer setting . This paper aims to provide a detailed review of the preclinical and clinical evidence relating to the mechanism of action , efficacy and safety of DB05773 for the treatment of P04626 -positive breast cancer . Systemic treatment of P04626 -positive metastatic breast cancer : a systematic review . AIM : We aimed to systematically review and summarize data from the available clinical trials that examined the treatment of P04626 -positive metastatic breast cancer . METHODS : We reviewed phase 2 and 3 studies in which an anti- P04626 agent was used in one or both arms of the study . While formal meta-analysis was not possible for such a heterogeneous group of trials , resulting forest plots outline some generalizable findings . RESULTS : There is strong evidence that the addition of an anti- P04626 agent to standard chemo- or endocrine therapy improves clinically relevant measurable outcomes . There is also consistent evidence that initial treatment with trastuzumab alone ( and subsequent use of a cytotoxic ) is inferior to the initial combination of trastuzumab plus chemotherapy , and that either DB05773 or dual anti- P04626 agents are superior to single anti- P04626 agent regimens . There is no strong evidence that the use of more than one cytotoxic agent together with an anti- P04626 agent confers any benefit over a single cytotoxic , anti- P04626 combination . CONCLUSION : This review provides a strong evidence base for current clinical practice with a discussion of treatment in the Australian setting . DB00072 emtansine : a unique antibody-drug conjugate in development for human epidermal growth factor receptor 2-positive cancer . DB00072 emtansine ( DB05773 ) is a human epidermal growth factor receptor ( P04626 ) -targeted antibody-drug conjugate , composed of trastuzumab , a stable thioether linker , and the potent cytotoxic agent DM1 ( derivative of maytansine ) , in phase III development for P04626 -positive cancer . Extensive analysis of DB05773 in preclinical studies has shown that DB05773 combines the distinct mechanisms of action of both DM1 and trastuzumab , and has antitumor activity in trastuzumab- and lapatinib-refractory experimental models . Clinically , DB05773 has a consistent pharmacokinetics profile and minimal systemic exposure to free DM1 , with no evidence of DM1 accumulation following repeated DB05773 doses . Although a few covariates were shown to affect interindividual variability in DB05773 exposure and clearance in population-pharmacokinetics analyses , the magnitude of their effect on DB05773 exposure was not clinically relevant . Phase I and phase II clinical trials of DB05773 as a single agent and in combination with paclitaxel , docetaxel , and pertuzumab have shown clinical activity and a favorable safety profile in patients with P04626 -positive metastatic breast cancer . Two randomized phase III trials of DB05773 are recruiting patients : EMILIA ( NCT00829166 ) is evaluating DB05773 compared with lapatinib plus capecitabine , and MARIANNE ( NCT01120184 ) is evaluating DB05773 plus placebo versus DB05773 plus pertuzumab versus trastuzumab plus a taxane . Additional combinations of DB05773 ( for example , with P16260 -0941 ) and additional disease settings ( early-stage P04626 -positive breast cancer ) are also under investigation . Data from the phase III trials and other studies of DB05773 -containing agents are eagerly awaited . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Responses to subsequent anti- P04626 therapy after treatment with trastuzumab-DM1 in women with P04626 -positive metastatic breast cancer . BACKGROUND : Women with human epidermal growth factor receptor 2 ( P04626 ) -positive metastatic breast cancer ( MBC ) can respond to multiple lines of anti- P04626 therapy . It is unknown whether these patients will derive further clinical benefit following treatment with trastuzumab-MCC-DM1 ( DB05773 ) . PATIENTS AND METHODS : We retrospectively identified P04626 -positive MBC patients treated with DB05773 and characterized outcomes during subsequent lines of anti- P04626 therapy . Response was determined by a blinded radiology review . Time-dependent analyses were carried out using Kaplan-Meier estimates . RESULTS : We identified 23 patients treated with single-agent DB05773 and report on the 20 patients who discontinued protocol therapy . All patients received trastuzumab-based metastatic therapy before initiation of DB05773 [ median 7 regimens ( range 3-14 ) ] . Of these 20 patients , 75 % ( 15 of 20 ) received further therapy with or without anti- P04626 agents after discontinuing DB05773 . Partial response to either first- or second-subsequent line(s) of therapy was seen in 5 of 15 ( 33 % ) treated patients , including 33 % ( 4 of 12 ) who received a regimen containing trastuzumab and/or lapatinib . Median durations of therapy to first- and second-subsequent regimens after DB05773 were 5.5 and 6.4 months , respectively . CONCLUSIONS : In heavily pretreated P04626 -positive MBC patients , prior exposure to DB05773 does not exhaust the potential benefit of ongoing anti- P04626 therapy with trastuzumab- and/or lapatinib-based regimens . Clinical implications of pathophysiological and demographic covariates on the population pharmacokinetics of trastuzumab emtansine , a P04626 -targeted antibody-drug conjugate , in patients with P04626 -positive metastatic breast cancer . DB00072 emtansine ( DB05773 ) is a P04626 -targeted antibody-drug conjugate in development for treatment of P04626 -positive cancers . DB05773 has been tested as a single agent in a phase I and 2 phase II studies of patients with heavily pretreated metastatic breast cancer ( MBC ) , with the maximum tolerated dose established at 3.6 mg/kg intravenously for every-3-week dosing . The authors present results from the population pharmacokinetics analysis for DB05773 . Population pharmacokinetics for DB05773 were characterized using a clinical database of 273 patients from the 3 studies . Pharmacokinetics was best described by a linear 2-compartment model . Population estimates ( interindividual variability [ IIV ] ) for pharmacokinetic parameters were clearance , 0.7 L/d ( 21.0 % ) ; central compartment volume ( V(c) ) , 3.33 L ( 13.2 % ) ; peripheral compartment volume ( V(p) ) , 0.89 L ( 50.4 % ) ; and intercompartmental clearance , 0.78 L/d . Body weight , albumin , tumor burden , and aspartate aminotransferase levels were identified as statistically significant covariates accounting for interindividual variability in DB05773 pharmacokinetics , with body weight having a greater effect on IIV of clearance and V(c) than other covariates . DB05773 exposure was relatively consistent across the weight range following body weight-based dosing . This analysis suggests no further DB05773 dose adjustments are necessary in heavily pretreated patients with MBC . Letter to the editor concerning ' DB00072 emtansine ( DB05773 ) versus lapatinib plus capecitabine in patients with P04626 -positive metastatic breast cancer and central nervous system metastases : a retrospective , exploratory analysis in EMILIA ' . 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 . Invasive Lobular Carcinomas Do Not Express Basal Cytokeratin Markers CK5/6 , CK14 and CK17 . The expression of basal cytokeratin markers CK5/6 in breast carcinomas has been associated with high histological grade and poor clinical outcome . A previous study has shown that CK5/6 can be detected in up to 17 % of invasive lobular carcinomas ( Q9Y4X3 ) . Here we study the expression of three basal cytokeratin markers ( CK5/6 , CK14 , and CK17 ) in 53 Q9Y4X3 cases diagnosed by histology and lack of P12830 expression . Among them , 42 were classic lobular carcinomas , 6 were tubular-lobular carcinoma , and 5 were pleomorphic lobular carcinomas . There was no significant difference among these three groups in patients ' age , tumor size , uni- and multi-focality , expression of ER and PR , lymphovascular invasion , perineural invasion and lymph node metastasis . The only statistically different factor was P04626 over-expression , which was observed only in pleomorphic Q9Y4X3 ( P = 0.0073 ) . None of the 53 cases expressed CK5/6 , CK14 or CK17 ; and 51/53 cases expressed luminal markers CK8 and CK18 , and the two negative cases were both classic lobular carcinoma , with positivity for ER and PR . In conclusion , all 53 cases of Q9Y4X3 failed to show expression by any of the three basal CK markers , suggesting that very few Q9Y4X3 will demonstrate a basal phenotype when assessed by immunohistochemistry ( IHC ) . More studies are needed to investigate molecular classification in lobular carcinoma of the breast . Survivin expression in breast cancer predicts clinical outcome and is associated with P04626 , P15692 , urokinase plasminogen activator and P05121 . BACKGROUND : Survivin , a novel inhibitor of apoptosis , is one of the most cancer-specific proteins identified to date . In this study we ( a ) evaluated the association between survivin and P04626 , vascular endothelial growth factor ( P15692 ) and uPA/ P05121 expression and ( b ) defined its effect on clinical outcome in a large breast cancer patient cohort . PATIENTS AND METHODS : Survivin expression was measured by ELISA in primary breast cancer tissue extracts from 420 patients with long-term clinical follow-up . RESULTS : Survivin was detected in 378 ( 90 % ) of the 420 primary breast cancer cases . Increased survivin levels were significantly associated with high nuclear grade ( P < 0.0001 ) , negative hormone receptor status ( P = 0.0028 ) , P04626 overexpression ( P = 0.0094 ) , P15692 expression ( P < 0.0001 ) , high uPA ( P = 0.0002 ) and P05121 levels ( P = 0.0002 ) . Using the 25th percentile ( 1.4 ng/mg ) as a cut-off point , patients expressing elevated survivin had a significantly worse disease-free survival ( DFS : P = 0.0007 , RR 1.97 ) and overall survival ( OS : P = 0.0009 , RR 2.11 ) compared with patients expressing lower levels of survivin . In multivariate analysis , this prognostic value of survivin was independent of both traditional and novel clinicopathologic factors for both DFS ( P = 0.0076 , RR 1.72 ) and OS ( P = 0.0155 , RR 1.76 ) . CONCLUSIONS : The independent prognostic relevance of survivin , when combined with previous data from model systems implicating survivin in the inhibition of apoptosis , suggests that survivin may be a suitable target for future therapeutic strategies . Safety and efficacy of the combination of DB05773 with radiotherapy of the central nervous system in a patient with P04626 -positive metastatic breast cancer : case study and review of the literature . Approximately 35 % of patients with confirmed P04626 breast cancer progress to metastases of the central nervous system ( CNS ) . Total cerebral radiotherapy is considered as standard treatment for these cases ; however , studies have shown that some chemotherapy drugs can be used during radiotherapy without significantly increasing its toxicity . In this article , we report the case of a patient with P04626 -positive breast cancer who showed isolated progression of the illness in the CNS , which was observed during the treatment period using DB05773 concomitantly with radiotherapy of the CNS without apparent toxicity of the combination and keeping the illness controlled . Through a review of the literature on the use of radiotherapy and chemotherapy with DB05773 for the treatment of cerebral metastases in P04626 -positive breast cancer , we describe the efficacy and tolerance of the concomitant application of these treatments . Updates on the treatment of human epidermal growth factor receptor type 2-positive breast cancer . PURPOSE OF REVIEW : To review the most recent developments in the treatment of human epidermal growth factor receptor type 2 ( P04626 ) -positive breast cancer with novel P04626 -targeting agents and combinations that have significantly improved clinical outcomes . RECENT FINDINGS : Since the approval of trastuzumab 15 years ago , the natural history of P04626 -positive breast cancer has been altered with improvements in survival for both early and advanced disease with the addition of this agent to standard chemotherapy . The P04626 receptor pathway drives breast cancer growth and aggressiveness , and P04626 -targeted agents can improve survival in early and advanced disease . In the advanced setting , two new drugs have been approved since 2012 , pertuzumab and ado-trastuzumab emtansine ( DB05773 ) , both of which improve survival without any reciprocal increase in toxicity . However , resistance almost always ensues , pointing to the need to understand the driving mechanisms and to biomarkers that will help individualize therapy and point to newer signal transduction and other modulators . SUMMARY : P04626 -positive breast cancer represents a distinct subtype with more aggressive clinical characteristics . P04626 -targeted therapies , usually in combination with chemotherapy , are the standard of care , improving the cure rate in early-stage breast cancer and lengthening survival in the advanced setting . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . Targeting the P04626 receptor in metastatic breast cancer . The advent of targeted therapies has revolutionized the treatment of certain types of cancer . Identification of molecular targets on cancer cells has led to the design of novel drugs , which either used as single agents or in combination with chemotherapy , has prolonged survival in metastatic disease , or contributed to curative treatment in the adjuvant setting . A literature review was conducted to identify and present current knowledge on the molecular function of the P04626 receptor , its role in the pathogenesis of breast cancer and anti- P04626 targeted drugs in use or under development . Many molecular targets have been identified in breast cancer , with the HER family of receptors being the ones most extensively studied . DB00072 and lapatinib target the P04626 receptor and are approved drugs for the treatment of metastatic breast cancer . Several other targeted agents , including DB05773 , pertuzumab , neratinib , afatinib and ertumaxomab , are currently being tested in vivo as well as in clinical studies . The use of targeted therapies in metastatic breast cancer has improved prognosis , increased survival and dramatically changed the way we treat breast cancer patients today . Estrogen regulation in human breast cancer cells of new downstream gene targets involved in estrogen metabolism , cell proliferation and cell transformation . We explored , by cDNA mini-arrays , gene expression measurements of MVLN , a human breast carcinoma cell line derived from MCF-7 , after 4 days of exposure to 17beta-estradiol ( E(2) ) treatment , in order to extend our understanding of the mechanism of the pharmacological action of estrogens . We focused on 22 genes involved in estrogen metabolism , cell proliferation regulation and cell transformation . The specificity of the E(2) response was reinforced by comparison with 4-hydroxytamoxifen ( OH-Tam ) , DB00947 and E(2)+OH-Tam expression profiles . Real-time quantitative PCR ( RTQ-PCR ) confirmed the variation of expression of known ( P04155 , P15514 , P35568 , P22692 , P12004 , P04626 , P07339 , MYC ) as well as novel ( DLEU2 , P20248 , P22309 , O15438 , O15440 , O75410 , P20827 , NOV , P01040 , P51511 , O75362 ) genes . The temporal response of these gene expression regulations was then investigated after 6 and 18 h of E(2) treatment and this allowed the identification of different time-course patterns . Cycloheximide treatment studies indicated first that estrogen affected the transcript levels of O15438 and O15440 through dissimilar pathways , and secondly that protein synthesis was needed for modulation of the expression of the P20248 and O75410 genes by estrogens . Western blot analysis performed on P04155 , P35568 , P22692 , amphiregulin , P12004 , cyclin A2 , O75410 and O15440 proteins confirmed the mini-array and RTQ-PCR data , even for genes harboring low variations of mRNA expression . Our findings should enhance the understanding of changes induced by E(2) on the transcriptional program of human E(2)-responsive cells and permit the identification of new potential diagnostic/prognostic tools for the monitoring of estrogen-related disease conditions such as breast cancer . Characterization of P04626 status by fluorescence in situ hybridization ( Q5TCZ1 ) and immunohistochemistry ( IHC ) . The use of human epidermal growth factor receptor type 2 ( P04626 ) gene amplification and overexpression as a molecular predictive marker has become critically important for proper selection of breast cancer patients for treatment with targeted therapeutic agents such as trastuzumab , lapatinib , pertuzumab , and DB05773 . A high level of sensitivity and specificity of molecular tests for this alteration is desirable . The American Society of Clinical Oncology and College of American Pathology have jointly established consensus guidelines to standardize characterization of this alteration in breast cancers . This chapter provides a brief overview of pre-analytic and analytical processing of breast specimens as well as subsequent molecular evaluation for P04626 status . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . DB00072 emtansine in human epidermal growth factor receptor 2-positive breast cancer : a review . PURPOSE OF REVIEW : In this review , we aim to update the clinical data of trastuzumab-DM1 ( DB05773 ) in terms of safety and efficacy , and describe ongoing and future trials evaluating its potential role in the management of patients with human epidermal growth factor receptor 2 ( P04626 ) -positive breast cancer . RECENT FINDINGS : DB00072 emtansine ( DB05773 ) is an antibody drug conjugate that optimizes delivery of chemotherapy with an anti- P04626 monoclonal antibody . As a conjugate , DB05773 's systemic side effects are significantly minimized due to its targeted delivery by trastuzumab to P04626 -positive cells . Phase I and II studies show that the maximum tolerated dose , and thus the recommended dose for DB05773 , is 3.6 mg/kg given intravenously every 3 weeks . Single arm phase Ib/II , II and a randomized phase II first-line study of DB05773 versus the combination of trastuzumab + docetaxel all showed improved tolerability , and at least equivalent efficacy , compared with our current standard of care . Two randomized phase III registration studies are now active , evaluating this agent in the refractory and first-line P04626 -positive settings . SUMMARY : DB05773 has been shown to be a very promising agent for the targeted delivery of chemotherapy and anti- P04626 monoclonal antibody therapy for patients with metastatic , P04626 -positive breast cancer . DB05773 will likely play a role in the management of patients with advanced and early stage P04626 -positive breast cancer , but this awaits further study . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Potential mechanisms for thrombocytopenia development with trastuzumab emtansine ( DB05773 ) . PURPOSE : DB00072 -emtansine ( DB05773 ) is an antibody-drug conjugate ( ADC ) comprising the cytotoxic agent DM1 conjugated to trastuzumab with a stable linker . Thrombocytopenia was the dose-limiting toxicity in the phase I study , and grade ≥3 thrombocytopenia occurred in up to 13 % of patients receiving DB05773 in phase III studies . We investigated the mechanism of DB05773 -induced thrombocytopenia . EXPERIMENTAL DESIGN : The effect of DB05773 on platelet function was measured by aggregometry , and by flow cytometry to detect the markers of activation . The effect of DB05773 on differentiation and maturation of megakaryocytes ( MK ) from human hematopoietic stem cells was assessed by flow cytometry and microscopy . Binding , uptake , and catabolism of DB05773 in MKs , were assessed by various techniques including fluorescence microscopy , scintigraphy to detect T-[H(3)]-DM1 and (125)I- DB05773 , and mass spectrometry . The role of FcγRIIa was assessed using blocking antibodies and mutant constructs of trastuzumab that do not bind FcγR . RESULTS : DB05773 had no direct effect on platelet activation and aggregation , but it did markedly inhibit MK differentiation via a cytotoxic effect . Inhibition occurred with DM1-containing ADCs but not with trastuzumab demonstrating a role for DM1 . MKs internalized these ADCs in a P04626 -independent , FcγRIIa-dependent manner , resulting in intracellular release of DM1 . Binding and internalization of DB05773 diminished as MKs matured ; however , prolonged exposure of mature MKs to DB05773 resulted in a disrupted cytoskeletal structure . CONCLUSIONS : These data support the hypothesis that DB05773 -induced thrombocytopenia is mediated in large part by DM1-induced impairment of MK differentiation , with a less pronounced effect on mature MKs . P35354 expression in prognosis and in prediction to endocrine therapy in early breast cancer patients . In breast cancer , the prognostic impact of P35354 expression varies widely between studies . We examined the prognostic value of P35354 expression in a large cohort of breast cancer patients treated with primary surgery between 1985 and 1994 and explained the variable results of P35354 expression found in the literature . A tissue microarray was constructed of available tumour material , and ER , PgR , P04626 , Ki67 and P35354 were examined by immunohistochemistry . Median follow-up was 19 years . Fifty-five percent ( n = 369/677 ) of patients received no systemic treatment . P35354 was scored using a weighted histoscore . Analysis of P35354 expression in two groups based on the median ( 148 ; below vs. above ) showed an increased hazard ratio ( HR ) of 1.35 ( 95 % CI 1.05-1.75 , P = 0.021 ) for disease-free survival ( DFS ) and of 1.39 ( 95 % CI 1.03-1.82 , P = 0.016 ) for overall survival ( OS ) . However , P35354 did not remain independent in multivariate analysis . In patients with hormone receptor positive tumours , P35354 expression had a negative influence on outcome ( low vs. high : DFS : HR 1.37 , 95 % CI 1.07-1.76 , P = 0.013 ) . This effect disappeared when endocrine therapy was administered ( low vs. high : DFS : HR 0.93 , 95 % CI 0.51-1.70 , P = 0.811 ) while it remained statistically significant when endocrine therapy was omitted ( low vs. high : DFS : HR 1.48 , 95 % CI 1.12-1.94 , P = 0.005 ) . Our results show that P35354 plays a role in hormonal pathways . Our results can explain the results found in previously published studies . P04626 -positive breast cancer : beyond trastuzumab . The outlook for patients with P04626 -positive breast cancer was revolutionized by the development of trastuzumab ( Herceptin ) , a humanized murine monoclonal antibody . Use of this agent led to improved overall survival when it was added to chemotherapy for the treatment of metastatic breast cancer . Improved understanding of mechanisms of resistance to trastuzumab has facilitated the development of novel agents for P04626 -positive breast cancer , and also resulted in superior outcomes when added to chemotherapy in the adjuvant setting . This review explores the use of several such agents , including lapatinib ( DB01259 ) , HSP90 inhibitors , DB05773 , and other tyrosine kinase inhibitors . Emerging data from trials of these agents indicate that the P04626 pathway remains a valid therapeutic target following disease progression on trastuzumab , and suggest a promising role for combined P04626 blockade with two or more agents . DB05773 : a P04626 -positive targeted antibody-drug conjugate . OBJECTIVE : To review the pharmacology , pharmacokinetics , efficacy , adverse effects , drug-drug interactions , dosage and administration , and formulary considerations for ado-trastuzumab emtansine . DATA SOURCES : Sources of information were identified through a PubMed search ( 1966 to June 2014 ) using the key terms ado-trastuzumab emtansine , trastuzumab-DM1 , trastuzumab-MCC-DM1 , and DB05773 . Other information was obtained from clinicaltrials.gov , product labeling , and press releases . STUDY SELECTION AND DATA EXTRACTION : All English-language clinical trials and abstracts evaluating ado-trastuzumab emtansine in humans were reviewed for inclusion . DATA SYNTHESIS : Overexpression or amplification of human epidermal growth factor receptor 2 ( P04626 ) occurs in approximately 20 % of breast cancers and is associated with more aggressive tumors and poorer prognosis in the absence of treatment . Although effective therapies for the initial management of P04626 -positive metastatic breast cancer ( MBC ) exist , many patients will experience disease progression . Most second-line therapies are associated with either significant toxicities or limited improvements in overall survival ( OS ) . DB05773 is a P04626 -positive directed antibody drug conjugate ( ADC ) approved in February 2013 . In phase III clinical trials comparing the efficacy and safety of ado-trastuzumab emtansine with lapatinib-capecitabine or physician 's choice , ado-trastuzumab emtansine had a better tolerability profile and improved progression-free survival compared with lapatinib-capecitabine or physician 's choice and increased OS compared with lapatinib-capecitabine . CONCLUSION : DB05773 is a novel ADC effective for P04626 -positive MBC in patients previously treated with trastuzumab , lapatinib , and a taxane . Further studies will determine its use in the adjuvant and neoadjuvant setting and in combination with pertuzumab . DB00877 effects transcriptional programs in smooth muscle cells controlling proliferative and inflammatory properties . Neointima formation , the leading cause of restenosis , is caused by proliferation of coronary artery smooth muscle cells ( CASMCs ) and is associated with infiltration by monocytes . DB00877 inhibits neointima formation after stent implantation in humans . It reduces proliferation by its effects on mammalian target of rapamycin ( P42345 ) kinase . In this study , we investigated the expression of P42345 in human neointima and the effect of rapamycin on global transcriptional events controlling CASMC phenotype . In neointimal CASMCs , P42345 exhibited increased phosphorylation and was translocated to the nucleus compared with control . Comparative gene expression analysis of CASMCs treated with rapamycin ( 100 ng/ml ) revealed down-regulation of the transcription factor Q01094 , a key regulator of G(1)/S-phase entry , and of various retinoblastoma protein/ Q01094 -regulated genes . In addition , we found changes in the expression of genes associated with replication , apoptosis , and extracellular matrix formation . Furthermore , rapamycin decreased the gene expression of endothelial monocyte-activating polypeptide-II ( EMAP-II ) . This decrease of EMAP-II expression was reflected in a reduced adhesiveness of CASMCs for monocytic cells . Addition of EMAP-II counteracted the antiadhesive effect of rapamycin . Therefore , EMAP-II may comprise a mechanism of rapamycin-mediated reduction of the proinflammatory activation of CASMCs . The effects reported here of rapamycin on the down-regulation of genes involved in cell cycle progression , apoptosis , proliferation , and extracellular matrix formation in CASMCs provide an explanation of how rapamycin reduces CASMC proliferation . In addition , rapamycin may contribute to a reduction of inflammatory responses by reducing the adhesiveness of CASMC , a mechanism suggested to be mediated by the production and release of EMAP II . P04626 -directed therapy for metastatic breast cancer . Human epidermal growth factor receptor 2 ( P04626 ) overexpression drives the biology of 20 % of breast cancers , and predicts a poor prognosis for patients . P04626 -targeted therapies significantly improve outcomes for P04626 -positive patients with both early and metastatic breast cancer . Currently three P04626 -targeted agents , trastuzumab ( Herceptin ) , lapatinib ( DB01259 ) , and pertuzumab ( Perjeta ) , are available for the treatment of P04626 -positive metastatic breast cancer ( MBC ) . Numerous studies have attempted to optimize their use by combining them with each other , or with endocrine and cytotoxic therapies . Most recently , the FDA approved the combination of trastuzumab , pertuzumab , and docetaxel as first-line treatment for MBC , and in late February 2013 approved a fourth P04626 -targeted agent , trastuzumab emtansine ( DB05773 , Kadcyla ) , for accelerated approval . These advances create a number of clinical dilemmas , including identification of the optimal sequence of P04626 -targeted agents and the best drug combinations to use , as well as the recognition of primary and acquired drug resistance . In this article , we review clinical data informing the effective management of P04626 -positive MBC . DB00072 emtansine in advanced human epidermal growth factor receptor 2-positive breast cancer . INTRODUCTION : DB00640 - trastuzumab emtansine ( DB05773 ) is a human epidermal growth factor receptor 2 ( P04626 ) -targeted antibody-drug conjugate composed of trastuzumab , a stable linker ( MCC ) , and the cytotoxic agent DM1 ( derivative of maytansine ; mertansine ) . DB05773 retains the mechanisms of action of trastuzumab , but also acts as a , selectively delivered , tubulin inhibitor . Following antigen-mediated binding to the tumor cell , DB05773 is endocytosed and intracellularly catabolized resulting in the release of its cytotoxic moiety . AREAS COVERED : DB05773 has completed Phase III development and compared favorably with the lapatinib/capecitabine combination with a superior response rate ( objective response rate [ ORR ] ) and duration of response , longer duration of disease control ( progression-free survival [ PFS ] ) , prolonged overall survival and improved tolerability and quality of life in patients with prior treatment with trastuzumab and a taxane . In a separate Phase III , DB05773 was compared with any other chosen regimen in patients who had at least received two prior P04626 -directed therapies . DB05773 nearly doubled PFS . EXPERT OPINION : DB05773 ( Kadcyla ) has become the treatment of choice in second-line and beyond for patients with advanced P04626 -expressing breast cancer . Dynamic microtubules regulate the local concentration of P12830 at cell-cell contacts . In contrast to the well-established relationship between cadherins and the actin cytoskeleton , the potential link between cadherins and microtubules ( MTs ) has been less extensively investigated . We now identify a pool of MTs that extend radially into cell-cell contacts and are inhibited by manoeuvres that block the dynamic activity of MT plus-ends ( e.g. in the presence of low concentrations of DB08313 and following expression of a P30622 mutant ) . Blocking dynamic MTs perturbed the ability of cells to concentrate and accumulate P12830 at cell-cell contacts , as assessed both by quantitative immunofluorescence microscopy and fluorescence recovery after photobleaching ( P42345 ) analysis , but did not affect either transport of P12830 to the plasma membrane or the amount of P12830 expressed at the cell surface . This indicated that dynamic MTs allow cells to concentrate P12830 at cell-cell contacts by regulating the regional distribution of P12830 once it reaches the cell surface . Importantly , dynamic MTs were necessary for myosin II to accumulate and be activated at cadherin adhesive contacts , a mechanism that supports the focal accumulation of P12830 . We propose that this population of MTs represents a novel form of cadherin-MT cooperation , where cadherin adhesions recruit dynamic MTs that , in turn , support the local concentration of cadherin molecules by regulating myosin II activity at cell-cell contacts . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . Population pharmacokinetics of trastuzumab emtansine ( DB05773 ) , a P04626 -targeted antibody-drug conjugate , in patients with P04626 -positive metastatic breast cancer : clinical implications of the effect of covariates . PURPOSE : DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate comprising the humanized monoclonal antibody trastuzumab linked to DM1 , a highly potent cytotoxic agent . A population pharmacokinetic ( PK ) analysis was performed to estimate typical values and interindividual variability of DB05773 PK parameters and the effects of clinically relevant covariates . METHODS : Serum samples were collected from 671 patients with human epidermal growth factor receptor 2-positive locally advanced or metastatic breast cancer ( MBC ) who received single-agent DB05773 in five phase I to phase III studies . Nonlinear mixed-effects modeling with the first-order conditional estimation method was used . RESULTS : A linear two-compartment model with first-order elimination from the central compartment described DB05773 PKs in the clinical dose range . DB05773 elimination clearance was 0.676 L/day , volume of distribution in the central compartment ( V c ) was 3.127 L , and terminal elimination half-life was 3.94 days . Age , race , region , and renal function did not influence DB05773 PK . Given the low-to-moderate effect of all statistically significant covariates on DB05773 exposure , none of these covariates is expected to result in a clinically meaningful change in DB05773 exposure . CONCLUSIONS : DB05773 PK properties are consistent and predictable in patients . A further refinement of dose based on baseline covariates other than body weight for the current 3.6 mg/kg regimen would not yield clinically meaningful reductions in interindividual PK variability in patients with MBC . DB02342 causes functional repression of transforming growth factor β3 signaling by ameliorating Smad and non-Smad signaling pathways in immortalized uterine fibroid cells . OBJECTIVE : To investigate the effects and the mechanism of action of 2-methoxyestradiol ( 2ME(2) ) on transforming growth factor ( TGF ) β3-induced profibrotic response in immortalized human uterine fibroid smooth muscle ( huLM ) cells . DESIGN : Laboratory study . SETTING : University research laboratory . PATIENTS(S) : Not applicable . INTERVENTIONS(S) : Not applicable . MAIN OUTCOME MEASURE(S) : huLM cells were treated with TGF-β3 ( 5 ηg/mL ) in the presence or absence of specific P84022 inhibitor SIS3 ( 1 μmol/L ) , inhibitor of the PI3K/Akt ( LY294002 , 10 μmol/L ) , or 2ME(2) ( 0.5 μmol/L ) , and the expression of collagen ( Col ) type I(αI) , Col III(αI) , plasminogen activator inhibitor ( P05121 ) 1 , connective tissue growth factor ( P29279 ) , and α-smooth muscle actin ( α-SMA ) were determined by real-time reverse-transcription polymerase chain reaction and immunoblotting . The effect of 2ME(2) on Smad-microtubule binding was evaluated by coimmunoprecipitation . RESULT(S) : Our data revealed that TGF-β3-induced fibrogenic response in huLM is mediated by both Smad-dependent and Smad-independent PI3K/Akt/ P42345 signaling pathways . 2ME(2) abrogates TGF-β3-induced expression of Col I(αI) , Col III(αI) , P05121 , P29279 , and α-SMA . Molecularly , 2ME(2) ameliorates TGF-β3-induced Q15796 /3 phosphorylation and nuclear translocation . In addition , 2ME(2) inhibits TGF-β3-induced activation of the PI3K/Akt/ P42345 pathway . CONCLUSION(S) : TGF-β3-induced profibrotic response in fibroid cells is mediated by Smad-dependent and Smad-independent PI3K/Akt/ P42345 pathways . 2ME(2) inhibits TGF-β3 profibrotic effects in huLM cells by ameliorating both Smad-dependent and Smad-independent signaling pathways . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . DB05773 , a novel antibody-drug conjugate , is highly effective against primary P04626 overexpressing uterine serous carcinoma in vitro and in vivo . Amplification of c-erbB2 has been reported in over 30 % of uterine serous carcinoma ( USC ) and found to confer poor survival because of high proliferation and increased resistance to therapy . In this study , we evaluated for the first time DB00072 emtansine ( DB05773 ) , a novel antibody-drug conjugate , against multiple epidermal growth factor receptor-2 ( P04626 ) -positive USC cells in vitro followed by developing a supportive in vivo model . Fifteen primary USC cell lines were assessed by immunohistochemistry ( IHC ) and flow cytometry for P04626 protein expression . C-erbB2 gene amplification was evaluated using fluorescent in situ hybridization . Sensitivity to DB05773 and trastuzumab ( T ) -induced antibody-dependent cell-mediated cytotoxicity was evaluated in 5-h chromium release assays . DB05773 and T cytostatic and apoptotic activities were evaluated using flow-cytometry-based proliferation assays . In vivo activity of DB05773 versus T in USC xenografts in SCID mice was also evaluated . High levels of P04626 protein overexpression and P04626 gene amplification were detected in 33 % of USC cell lines . DB05773 was considerably more effective than trastuzumab in inhibiting cell proliferation and in causing apoptosis ( P = 0.004 ) of USC showing P04626 overexpression . Importantly , DB05773 was highly active at reducing tumor formation in vivo in USC xenografts overexpressing P04626 ( P = 0.04 ) and mice treated with TDM-1 had significantly longer survival when compared to T-treated mice and control mice ( P ≤ 0.0001 ) . DB05773 shows promising antitumor effect in P04626 -positive USC cell lines and USC xenografts and its activity is significantly higher when compared to T . DB05773 may represent a novel treatment option for P04626 -positive USC patients with disease refractory to trastuzumab and traditional chemotherapy . [ DB05773 and pertuzumab : emerging anti- P04626 therapeutics ] . P04626 -targeted therapy for P04626 -positive breast cancer is one of the success stories in medical oncology . DB00072 , a humanized monoclonal antibody , was the first approved P04626 -targeted agent . Subsequent developments include agents with different mechanisms , such as lapatinib , a tyrosine kinase inhibitor . We describe here the results of late-phase clinical trials of two newly-available anti- P04626 agents , DB05773 and pertuzumab . [ Antibody-drug conjugates in oncology : from the concept to trastuzumab emtansine ( DB05773 ) ] . DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate ( ADC ) which associates the selective intracellular targeting of the cytotoxic agent , DM1 ( maytansine derivative ) to the antitumor activity of trastuzumab . DB05773 targets the epidermal growth factor receptor 2 ( P04626 ) , highly expressed in the most aggressive forms of breast cancer . Current standard of care in P04626 -positive advanced or metastatic breast cancers has its limitations , particularly after progression on P04626 -targeted approved therapies . DB05773 showed a significant antitumor activity in vitro and in vivo , and in experimental models resistant to P04626 -targeted agents . Phase I and II studies showed that the maximum tolerated dose for DB05773 is 3.6 mg/kg given intravenously every three weeks . At this recommended dose , DB05773 provided objective tumor responses and favourable safety profile . A phase II randomised study , evaluating DB05773 in first line vs trastuzumab plus docetaxel , the current standard of care in advanced or metastatic breast cancers , showed improved tolerability and efficacy . Recently , the results of EMILIA , a phase III randomised study assessing , after prior treatment with trastuzumab and a taxane , the efficacy and the safety of DB05773 vs lapatinib plus capecitabine , confirmed the therapeutic benefit . DB05773 appears to be an effective therapeutic option to treat patients with P04626 -positive metastatic breast cancer . DB00072 emtansine ( DB05773 ) : a novel agent for targeting P04626 + breast cancer . Increased understanding of the molecular mechanisms of tumorigenesis has led to the development of novel agents that target tumor cells with minimal effects on normal cells . The success of this approach is exemplified by the development of monoclonal antibodies directed toward antigens expressed selectively by tumor cells . The conjugation of these monoclonal antibodies with potent cytotoxic drugs has the potential to further improve efficacy while retaining a favorable safety profile . DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate ( ADC ) currently in clinical development . It combines the humanized antibody trastuzumab , which targets the human epidermal growth factor receptor 2 ( P04626 ) receptor on cancer cells , and the potent antimicrotubule agent DM1 using a unique highly stable linker . When DB05773 binds to P04626 , a proportion of the receptors are thought to be internalized by the process of receptor endocytosis , followed by the intracellular release of an active form of DM1 , which in turn kills the tumor cell . This review presents the rationale for the development of DB05773 and summarizes the preclinical and clinical data for this novel agent for the treatment of breast cancer . OSU-03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier : Implications for Anti-Cancer Therapies . We examined the interaction between OSU-03012 ( also called AR-12 ) with phosphodiesterase 5 ( O76074 ) inhibitors to determine the role of the chaperone glucose-regulated protein ( P11021 ) / P11021 / P11021 in the cellular response . DB00203 ( Viagra ) interacted in a greater than additive fashion with OSU-03012 to kill stem-like GBM cells . Treatment of cells with OSU-03012/sildenafil : abolished the expression of multiple oncogenic growth factor receptors and plasma membrane drug efflux pumps and caused a rapid degradation of P11021 and other HSP70 and HSP90 family chaperone proteins . Decreased expression of plasma membrane receptors and drug efflux pumps was dependent upon enhanced Q9NZJ5 -eIF2α- P18848 - P35638 signaling and was blocked by P11021 over-expression . In vivo OSU-03012/sildenafil was more efficacious than treatment with celecoxib and sildenafil at killing tumor cells without damaging normal tissues and in parallel reduced expression of P08183 and Q9UNQ0 in the normal brain . The combination of OSU-03012/sildenafil synergized with low concentrations of sorafenib to kill tumor cells , and with lapatinib to kill P00533 over-expressing tumor cells . In multiplex assays on plasma and human tumor tissue from an OSU-03012/sildenafil treated mouse , we noted a profound reduction in uPA signaling and identified FGF and P23458 /2 as response biomarkers for potentially suppressing the killing response . Inhibition of FGFR signaling and to a lesser extent P23458 /2 signaling profoundly enhanced OSU-03012/sildenafil lethality . Human epidermal growth factor receptor 2 positive ( P04626 + ) metastatic breast cancer : how the latest results are improving therapeutic options . Human epidermal growth factor receptor 2 positive ( P04626 + ) metastatic breast cancer ( MBC ) remains an incurable disease , and approximately 25 % of patients with P04626 + early breast cancer still relapse after adjuvant trastuzumab-based treatment . P04626 is a validated therapeutic target that remains relevant throughout the disease process . Recently , a number of novel P04626 targeted agents have become available , including lapatinib ( a small molecule tyrosine kinase inhibitor of both P04626 and the epidermal growth factor receptor ) , pertuzumab ( a new anti- P04626 monoclonal antibody ) and ado-trastuzumab emtansine ( DB05773 , a novel antibody-drug conjugate ) , which provide additional treatment options for patients with P04626 + MBC . The latest clinical trials have demonstrated improved outcome with treatment including pertuzumab or DB05773 compared with standard P04626 targeted therapy . Here we review the clinical development of approved and investigational targeted agents for the treatment of P04626 + MBC , summarize the latest results of important clinical trials supporting use of these agents in the treatment of P04626 + MBC , and discuss how these results impact therapeutic options in clinical practice . Exposure-response relationship of DB05773 : insight into dose optimization for patients with P04626 -positive metastatic breast cancer . Exposure-response ( E-R ) analyses for ado-trastuzumab emtansine ( DB05773 , Kadcyla ) were performed using data from a randomized , active control ( lapatinib plus capecitabine ) trial in patients with human epidermal growth factor 2-positive metastatic breast cancer . Kaplan-Meier survival analyses stratified by DB05773 trough concentration on day 21 of cycle 1 ( Cmin,C1D21 ) were performed for overall survival ( OS ) and progression-free survival ( PFS ) . E-R analyses indicated that after adjusting for baseline risk factors , higher DB05773 exposure is associated with improved efficacy . DB05773 -treated patients with Cmin,C1D21 lower than the median value had values of OS and PFS comparable to those of the active control arm . The percentage of patients who received DB05773 dose adjustments was similar across the exposure range and was lower than that of the active control arm . Our findings suggest that there may be an opportunity to optimize Kadcyla dose in the patient subgroup with low DB05773 exposure for improved efficacy with acceptable tolerability . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors . Why your preferred targeted drugs may become unaffordable . DB00072 , a monoclonal antibody directed at the P04626 receptor , is one of the most impressive targeted drugs developed in the last two decades . Indeed , when given in conjunction with chemotherapy , it improves the survival of women with P04626 positive breast cancer , both in advanced and in early disease . Its optimal duration , however , is poorly defined in both settings with a significant economic impact in the adjuvant setting where the drug is arbitrarily given for 1 year . This article reviews current attempts at shortening this treatment duration , emphasizing the likelihood of inconclusive results and , therefore , the need to investigate this important variable as part of the initial pivotal trials and with the support of public health systems . Failure to do so has major consequences on treatment affordability . Ongoing adjuvant trials of dual P04626 blockade , using trastuzumab in combination with a second anti- P04626 agent , and trials of the antibody-drug conjugate DB05773 ( trastuzumab-emtansine ) have to all be designed with 12 months of targeted therapy . Cell cycle arrest in Metformin treated breast cancer cells involves activation of AMPK , downregulation of cyclin D1 , and requires P46527 or p21Cip1 . BACKGROUND : The antihyperglycemic drug metformin may have beneficial effects on the prevention and treatment of cancer . Metformin is known to activate AMP-activated protein kinase ( AMPK ) . It has also been shown to inhibit cyclin D1 expression and proliferation of some cultured cancer cells . However , the mechanisms of action by which metformin mediates cell cycle arrest are not completely understood . RESULTS : In this study , metformin was found to inhibit proliferation of most cultured breast cancer cell lines . This was independent of estrogen receptor , P04626 , or p53 status . Inhibition of cell proliferation was associated with arrest within G0/ P55008 phase of the cell cycle . As in previous studies , metformin treatment led to activation of ( AMPK ) and downregulation of cyclin D1 . However , these events were not sufficient for cell cycle arrest because they were also observed in the MDA-MB-231 cell line , which is not sensitive to growth arrest by metformin . In sensitive breast cancer lines , the reduction in cyclin D1 led to release of sequestered CDK inhibitors , P46527 and p21Cip1 , and association of these inhibitors with cyclin E/ P24941 complexes . The metformin-resistant cell line MDA-MB-231 expresses significantly lower levels of P46527 and p21Cip1 than the metformin-sensitive cell line , MCF7 . When P46527 or p21Cip1 were overexpressed in MDA-MB-231 , the cells became sensitive to cell cycle arrest in response to metformin . CONCLUSION : Cell cycle arrest in response to metformin requires CDK inhibitors in addition to AMPK activation and cyclin D1 downregulation . This is of interest because many cancers are associated with loss or downregulation of CDK inhibitors and the results may be relevant to the development of anti-tumor reagents that target the AMPK pathway . Beyond trastuzumab and lapatinib : new options for P04626 -positive breast cancer . P04626 -positive breast cancer ( BC ) constitutes a molecular subtype of the disease with an aggressive biologic behavior . DB00072 revolutionized the treatment of this disease , changing its natural history . DB01259 is active in the metastatic setting , approved for patients who were pretreated with trastuzumab . However , resistance to anti- P04626 agents is a major clinical issue , occurring in both early-stage and advanced disease , and new treatment options are clearly needed . An abundance of P04626 -targeted agents are being clinically developed : monoclonal antibodies , small molecule inhibitors , and antibody drug conjugates ( ADC ) . Combining P04626 -targeted agents in regimens of dual P04626 blockade has already reached clinical practice in the metastatic setting , confirming the preclinical efficacy of enhanced P04626 inhibition . Promising results have been generated in the neoadjuvant setting , and large randomized trials are seeking evidence for dual P04626 blockade in the adjuvant setting . ADC represent another hope for improved treatment outcomes of P04626 -positive BC , as exemplified by the positive results of clinical trials employing trastuzumab-DM1 ( trastuzumab emtansine , DB05773 ) . Moreover , an understanding of the molecular mechanisms mediating resistance to P04626 blockade has opened new therapeutic avenues , with several targeted agents entering clinical trials . This paper presents the clinical data of the P04626 -targeted agents under development , as well as an overview of the biologic rationale for the development of agents aimed at circumventing anti- P04626 resistance . 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 . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . DB00072 emtansine in breast cancer . INTRODUCTION : DB00072 emtansine ( DB05773 ) is a human epidermal growth factor receptor 2 ( P04626 ) -targeted antibody-drug conjugate ( ADC ) composed of trastuzumab , a stable linker ( MCC ) , and the cytotoxic agent DM1 ( derivative of maytansine ) . Administration of DB05773 leads to limited systemic exposure of free DM1 , with no evidence of DM1 accumulation after repeated dosing . AREAS COVERED : Phase I and Phase II clinical trials with DB05773 as a single agent and in combination with paclitaxel , docetaxel , and pertuzumab have shown substantial clinical activity and a favorable safety profile . A randomized , open-label , first-line trial comparing trastuzumab and docetaxel with single agent DB05773 showed a significant improved progression-free survival for DB05773 . EXPERT OPINION : DB05773 has successfully completed second-line Phase III development for advanced P04626 -positive breast cancer . The Phase III EMILIA study demonstrated an overall survival benefit for DB05773 compared to the combination of lapatinib and capecitabine in taxane-trastuzumab pretreated patients . DB05773 may offer delivery on a personalized basis of very potent cytotoxic agents in a cellular selective manner . Treatment of P04626 -overexpressing breast cancer . The HER family of receptors consists of four closely related type 1 transmembrane TK receptors : P00533 ( P00533 ) , P04626 , P21860 and Q15303 . Signalling via the HER family of receptors underpins the majority of the intricate array of cellular activities on which cell survival and functionality depend . Aberrant P04626 expression and/or functionality have been implicated in the evolution of breast cancer and this receptor has proved to be a potent target for anticancer therapies , including antibody-based therapies to prevent ligand binding , dimer formation or the recruitment of antibody-dependent cell-mediated cytotoxicity , and direct kinase inhibition to prevent molecular activation and recruitment of downstream signalling partners . Novel strategies against P04626 include HER tyrosine kinase inhibitors , HSP90 inhibitors and antibody-chemotherapy conjugates . This latter approach is exemplified by DB05773 , a potent antibody that has a good safety profile and that has shown remarkable activity in patients with advanced disease . In addition , pertuzumab , an mAb that directly inhibits the formation of P04626 dimers including the P04626 : P21860 dimer , offers a unique mechanism of P21860 inhibition . All these approaches have shown substantial clinical activity in patients refractory to trastuzumab . It is anticipated that with the increased availability of novel anti- P04626 agents together with a better understanding of the mechanisms of resistance to anti- P04626 agents we should be able to further improve the outcome of patients with P04626 breast cancer . There will also be an increasing tendency towards moving the study of these agents to earlier stages of the disease , namely in the adjuvant and neoadjuvant setting . [ Chemotherapy for breast cancer refractory to anthracycline , taxane or trastuzumab ] . Anthracycline , taxane or trastuzumab play a central role in systemic chemotherapy for breast cancer . The standard of subsequent treatment is capecitabine , S-1 , vinorelbine , irinotecan or gemcitabine . DB04845 or nanoparticle paclitaxel is effective for taxane-resistant breast cancer . DB01259 proves effective for trastuzumab-resistant P04626 -overexpressing breast cancer and also for brain metastasis . DB05773 , pertuzumab and neratinib are promising drugs . In terms of antiangiogenic agents , bevacizumab in combination with taxane demonstrates efficacy . DB06626 , sunitinib or pazopanib is under investigation . It is necessary to study the best manner of sequence and combination in these drugs . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Breast cancer brain metastases responding to lapatinib plus capecitabine as second-line primary systemic therapy . Brain metastases ( BM ) are diagnosed in up to 40 % of P04626 -positive breast cancer patients . Standard treatment includes local approaches such as whole-brain radiotherapy ( WBRT ) , radiosurgery , and neurosurgery . The landscape trial established primary systemic therapy as an effective and safe alternative to WBRT in selected patients with Her2-positive BM . We aim to further focus on the role of systemic therapy in oligosymptomatic patients by presenting this case report . We report on a 50-year-old patient diagnosed with multiple BM 5 years after early breast cancer diagnosis . As the patient was asymptomatic and had a favorable diagnosis-specific P02724 score , she received primary systemic treatment with DB05773 . She achieved partial remission within the brain for eight treatment cycles and then progressed despite stable extracranial disease . As the patient remained asymptomatic and refused WBRT , we decided upon trastuzumab , lapatinib plus capecitabine as second-line therapy . Another partial remission of BM was observed ; to date , she has received 11 treatment cycles without any sign of disease progression . In this case , WBRT was delayed by at least 14 months , again indicating the activity of systemic treatment in BM . Apparently , in selected patients , BM can be controlled with multiple lines of systemic therapy similar to extracranial disease . Further investigation of systemic treatment approaches is therefore warranted . Dual targeting of P04626 -positive cancer with trastuzumab emtansine and pertuzumab : critical role for neuregulin blockade in antitumor response to combination therapy . PURPOSE : Targeting P04626 with multiple P04626 -directed therapies represents a promising area of treatment for P04626 -positive cancers . We investigated combining the P04626 -directed antibody-drug conjugate trastuzumab emtansine ( DB05773 ) with the P04626 dimerization inhibitor pertuzumab ( Perjeta ) . EXPERIMENTAL DESIGN : Drug combination studies with DB05773 and pertuzumab were performed on cultured tumor cells and in mouse xenograft models of P04626 -amplified cancer . In patients with P04626 -positive locally advanced or metastatic breast cancer ( mBC ) , DB05773 was dose-escalated with a fixed standard pertuzumab dose in a 3+3 phase Ib/II study design . RESULTS : Treatment of P04626 -overexpressing tumor cells in vitro with DB05773 plus pertuzumab resulted in synergistic inhibition of cell proliferation and induction of apoptotic cell death . The presence of the P21860 ligand , heregulin ( Q99988 β ) , reduced the cytotoxic activity of DB05773 in a subset of breast cancer lines ; this effect was reversed by the addition of pertuzumab . Results from mouse xenograft models showed enhanced antitumor efficacy with DB05773 and pertuzumab resulting from the unique antitumor activities of each agent . In patients with mBC previously treated with trastuzumab , lapatinib , and chemotherapy , DB05773 could be dosed at the maximum tolerated dose ( MTD ; 3.6 mg/kg every 3 weeks ) with standard dose pertuzumab . Adverse events were mostly grade 1 and 2 , with indications of clinical activity . CONCLUSIONS : Dual targeting of P04626 with the combination of DB05773 and pertuzumab in cell culture and mouse xenograft models resulted in enhanced antitumor activity . In patients , this combination showed an encouraging safety and tolerability profile with preliminary evidence of efficacy .

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Molecular basis of functional gastrointestinal disorders . There are a number of abnormalities of gastrointestinal function , including sensory and motor dysfunction , which are believed to play a role in the manifestation of symptoms in patients with functional gastrointestinal disorders ( FGID ) . In addition , there is a remarkable psychiatric comorbidity . Family and twin studies have provided strong evidence for a clustering of FGID in families and an increased concordance in monozygotic compared to dizygotic twins . This points towards the role of one or more hereditary ( genetic ) factors . Considering these disorders of function and the psychiatric comorbidity , polymorphisms of adrenergic , opioidergic or serotonergic receptors as well as G-protein beta3 ( P16520 ) subunit gene polymorphisms ( C825T ) and polymorphisms of 5-HT transporter genes are suitable causes . In addition , mediators or regulators of mucosal inflammation may trigger events that ultimately result in the manifestation of FGID . Thus , relevant polymorphisms of genes with immunmodulating and/or neuromodulating features ( P35372 , P05112 , IL-4R , TNFalpha ) may also play a role in the manifestation of FGIDs . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . mu-Opioid receptor agonists differentially regulate the expression of miR-190 and Q13562 . The agonists of mu-opioid receptor ( P35372 ) induce extracellular signal-regulated kinase ( P29323 ) phosphorylation through different pathways : morphine uses the protein kinase C ( PKC ) -pathway , whereas fentanyl functions in a beta-arrestin2-dependent manner . In addition , the two pathways result in the different cellular location of phosphorylated P29323 and the activation of different sets of transcriptional factors . In the current study , the influence of the two pathways on the expression of microRNAs ( miRNAs ) was investigated . After treating the primary culture of rat hippocampal neurons and the mouse hippocampi with morphine or fentanyl for 3 days , seven miRNAs regulated by one or two of the agonists were identified . One of the identified miRNAs , miR-190 , was down-regulated by fentanyl but not by morphine . This down-regulation was attenuated by 1,4-diamino-2,3-dicyano-1,4-bis(methylthio)butadiene ( U0126 ) , which blocks the phosphorylation of P29323 . When fentanyl-induced but not morphine-induced P29323 phosphorylation was blocked in the primary cultures from beta-arrestin2(-/-) mouse , fentanyl did not decrease the expression of miR-190 . However , a PKC inhibitor that blocked morphine-induced P29323 phosphorylation specifically had no effect on the miR-190 down-regulation . Therefore the decrease in miR-190 expression resulted from the agonist-selective P29323 phosphorylation . In addition , the expressional changes in one of the miR-190 targets , neurogenic differentiation 1 ( Q13562 ) , correlated with those in miR-190 expression , suggesting the P35372 could regulate the Q13562 pathways via the control of miR-190 expression . A vitamin A deficient diet enhances proinflammatory cytokine , P35372 , and HIV-1 expression in the HIV-1 transgenic rat . The HIV-1 ( HIV ) transgenic ( Tg ) rat develops several immune abnormalities in association with clinical impairments that are similar to what are seen with HIV infection in humans . In HIV infection , retinoids and opioids can have separate and potentially combined effects on the clinical course of HIV disease . In these studies , the effects of a vitamin A deficient diet on T cell proinflammatory cytokine and mu opioid receptor ( MOR ) expression were examined in the Tg and in wild-type ( WT ) rats . The effects of the diet on HIV gene expression were also analyzed in the Tg rats . Phytohemagglutinin-stimulated T cells from WT rats on the vitamin A diet and from Tg rats on either diet were more likely to either produce increased percentages of T cells expressing intracytoplasmic P01579 , secrete higher levels of P01375 , and express higher levels of MOR mRNA and surface MOR . Mitogen stimulation also increased Tg rat HIV env , tat , and nef mRNA expression with even higher env and nef mRNA produced in association with the vitamin A deficient diet . All together , these data suggest that a vitamin A deficient diet can result in cellular effects that increase T cell proinflammatory responses and HIV expression , which may alter the course of disease in the HIV Tg rat model . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . DB00588 -induced regulation of the balance within macrophage subpopulations . In asthma , treatment with inhaled corticosteroids reduces chronic peribronchial inflammation and restores the balance within macrophage subpopulations . This study investigates whether corticosteroids can regulate monocyte differentiation in vitro and thereby influence the balance of functionally distinct macrophages . Graded doses of fluticasone propionate ( FP ) were added to cultures of normal peripheral blood monocytes in the presence or absence of P05112 . Cells were harvested after 7 days ' culture . Double immunofluorescence studies were performed on cytospins of differentiated macrophages using the MoAbs RFD1 and RFD7 to distinguish inductive and suppressive macrophages by their respective phenotypes . Macrophage function was determined by quantifying allostimulation in a mixed leucocyte reaction and by measuring tumour necrosis factor-alpha ( P01375 ) production . FP reduced the number of mature cells with a D1+ antigen-presenting phenotype and up-regulated the development of cells with the D1/D7+ and D7+ phenotypes . Functionally , this was associated with reduced stimulation of T cell proliferation in a mixed leucocyte reaction ( P08235 ) . DB00588 also reversed the increase in both D1+ expression and P01375 production induced by P05112 . The effect of FP persisted for 24 h after removal of FP from the culture medium . These results suggest that FP treatment of asthmatics may have a direct beneficial effect by normalizing the macrophage subset imbalance that contributes to the chronic peribronchial inflammation present in this condition . Colocalization and shared distribution of endomorphins with DB05875 , calcitonin gene-related peptide , gamma-aminobutyric acid , and the mu opioid receptor . The endomorphins are endogenous opioids with high affinity and selectivity for the mu opioid receptor ( MOR , P35372 , MOP ) . Endomorphin-1 ( DB00135 -Pro- DB00150 - DB00120 -NH(2) ; EM1 ) and endomorphin-2 ( DB00135 -Pro- DB00120 - DB00120 -NH(2) ; EM2 ) have been localized to many regions of the central nervous system ( CNS ) , including those that regulate antinociception , autonomic function , and reward . Colocalization or shared distribution ( overlap ) of two neurotransmitters , or a transmitter and its cognate receptor , may imply an interaction of these elements in the regulation of functions mediated in that region . For example , previous evidence of colocalization of EM2 with DB05875 ( SP ) , calcitonin gene-related peptide ( P80511 ) , and MOR in primary afferent neurons suggested an interaction of these peptides in pain modulation . We therefore investigated the colocalization of EM1 and EM2 with SP , P80511 , and MOR in other areas of the CNS . EM2 was colocalized with SP and P80511 in the nucleus of the solitary tract ( P30990 ) and with SP , P80511 and MOR in the parabrachial nucleus . Several areas in which EM1 and EM2 showed extensive shared distributions , but no detectable colocalization with other signaling molecules , are also described . P03372 -immunoreactive neurons contain calcitonin gene-related peptide , methionine-enkephalin or tyrosine hydroxylase in the female rat preoptic area . We have shown in our previous studies that estrogen treatment selectively influences calcitonin gene-related peptide ( P80511 ) - , methionine-enkephalin ( DB00134 -Enk ) - and tyrosine hydroxylase ( TH ) -immunoreactive ( IR ) intensities in the neurons of the periventricular preoptic nucleus ( Q9H237 ) and the medial preoptic area ( DB00603 ) of the female rat . In the present study , we examined whether estrogen receptor ( ER ) -IR neurons in the Q9H237 and DB00603 contain P80511 , DB00134 -Enk , or TH using a double-labeling immunohistochemical method and investigated changes in the number of double-labeling cells upon treatment with estrogen . Brain sections of ovariectomized rats and ovariectomized and estrogen-treated rat were stained using the avidin-biotin-peroxidase complex method followed by the peroxidase-anti-peroxidase method . The sections were first incubated with an anti-ER antibody in conjunction with nickel diaminobenzidine which produces a dark blue reaction product in the nucleus . Subsequently , P80511 , DB00134 -Enk or TH antisera were applied to these sections and the resulting brown diaminobenzidine reaction product in the cytoplasm was examined . Neurons that were double-labeled for ER and P80511 , DB00134 -Enk or TH were investigated in the Q9H237 and DB00603 . The number of doubly labeled ER/ P80511 - and ER/TH-IR neurons was large , whereas the number of ER/ DB00134 -Enk-IR neurons was small . These results suggest that ER in the Q9H237 and DB00603 may be more closely related to the mechanism of changes in P80511 - and TH-IR intensities upon estrogen treatment than that in DB00134 -Enk-IR intensity . Preliminary evidence of ethnic divergence in associations of putative genetic variants for methamphetamine dependence . Research into the biological processes that increase susceptibility to methamphetamine dependence has been conducted primarily in Asian populations . Using a case-control design this study 's purpose was to explore , among a population of methamphetamine-dependent Caucasians , six putative single nucleotide polymorphisms previously found to be associated with methamphetamine dependence in Asian populations . A total of 193 non-psychotic males ( 117 methamphetamine-dependent and 76 controls ) were genotyped for variants located in six genes ( P31749 , P32121 , P23560 , P21964 , P09211 , P35372 ) . Genotypic and allelic frequencies , odds ratios , and 95 % confidence intervals were calculated . None of the putative gene associations was significantly replicated in our sample of Caucasian men . Effect size comparisons suggest a trend toward allelic divergence for arrestin beta 2 ( P32121 ) and glutathione S-transferase P1 ( P09211 ) and allelic convergence for brain-derived neurotrophic factor ( P23560 ) . Results provide preliminary support for further exploration and validation of candidate single nucleotide polymorphisms ( SNPs ) for methamphetamine ( METH ) dependence reported among Asian populations across other ethnic/ancestral groups . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . Linkage assignment of eleven genes to the porcine genome . We report comparative linkage mapping of eleven genes in the swine genome by RFLP analysis . These genes include : Acid phosphatase type 5 ( P13686 ) , Cholecystokinin Type B Receptor ( P32239 ) , Antibiotic Peptide ( P49913 ) , P01308 -like Growth Factor 1 Receptor ( P08069 ) , Integrin Alpha M ( P11215 ) , Integrin Beta 2 ( ITGbeta2 ) , Opioid Receptor Mu-1 ( P35372 ) , Pro-hormone Converter ( PC1/3 ) , DB00162 Binding Q12988 ( P10745 ) , Ribosomal DNA ( RNR1 ) , and Zona Pellucida Glycoprotein 1 ( P60852 ) . The P32239 and ITGbeta2 loci define the ends of the linkage groups on Chromosomes ( Chro ) ( SSC ) 9p and 13qter , respectively . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5-fluoruracil-based chemotherapy regimens was analyzed for variants of IL-1B , P60568 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 and P01375 genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein .

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P25021 overexpression induces U937 cell differentiation despite triggered mechanisms to attenuate DB02527 signalling . Knowing that cell-surface receptors that recognize and respond to extracellular stimuli are key components for the regular communication between individual cells required for the survival of any living organism , the aim of the present work was to investigate the effect of P25021 overexpression on the U937 signal transduction pathway and its consequences on cell proliferation and differentiation . The overexpression of P25021 led to an increase in DB02527 basal levels , a leftward shift of agonist concentration-response curves , and similar maximal response to agonist treatment , suggesting that overexpressed H2Rs act as functional spare receptors . In this system cells triggered several mechanisms tending to restore DB02527 basal levels to those of the naïve cells . P25021 overexpression induced PDE activity stimulation and P25098 overexpression . In spite of the onset of these regulatory mechanisms , H2 agonist and rolipram treatments induced the terminal differentiation of the P25021 overexpressed clone , conversely to the naïve cells . Present findings show that stably P25021 overexpression alters DB02527 signalling as the result of not only the amounts of second messenger generated but also the activation or upregulation of various components of signalling cascade , leading to an adapted biologically unique system . This adaptation may represent an advantage or a disadvantage , depending on the biological system , but in any case , the existence of compensatory mechanisms should be considered when a clinical treatment is designed . Genetic mechanism of aspirin-induced urticaria/angioedema . PURPOSE OF REVIEW : DB00945 -induced urticaria/angioedema is a major aspirin-related hypersensitivity often associated with aspirin-intolerant asthma . Genetic studies on aspirin-intolerant asthma have shown chronic overproduction of cysteinyl leukotrienes . The genetic analysis of aspirin-induced urticaria/angioedema is limited , however . RECENT FINDINGS : A recent study on HLA genotypes has suggested that the HLA alleles DRB11302 and DQB10609 may be genetic markers for aspirin-induced urticaria/angioedema . A polymorphism study that examined nine single-nucleotide polymorphisms of five leukotriene-related genes [ P09917 ( encoding P09917 ) , P20292 ( P09917 -activating protein ) , P35354 ( cyclooxygenase 2 ) , Q16873 ( leukotriene C4 synthase ) , and Q9Y271 ( cysteinyl leukotriene receptor 1 ) ] found that promoter polymorphisms of P09917 ( -1708A > G ) and Q9Y271 ( -634C > T ) were significantly different between aspirin-intolerant asthma and aspirin-induced urticaria/angioedema , suggesting different contributions to the lipoxygenase pathway . A second polymorphism study , conducted on histamine-related genes , did not find any significant associations with aspirin-induced urticaria/angioedema for the genes P50135 ( encoding histamine N-methyltransferase ) , P35367 or P25021 ( encoding histamine receptor types 1 and 2 respectively ) , or the gene encoding high-affinity IgE receptor Ibeta ( FcepsilonRIbeta ) ; however , the FcepsilonRIalpha gene promoter polymorphism was significantly associated with aspirin-induced urticaria/angioedema . This finding has been supported by in vitro functional studies . SUMMARY : The HLA alleles DRB11302 and DQB10609 , and the P09917 and FcepsilonRIalpha promoter polymorphisms , may contribute to the pathogenesis of aspirin-induced urticaria/angioedema . Further investigation to identify candidate genetic markers would help to elucidate the pathogenic mechanism of this condition . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Examination of ceramic restorative material interfacial debonding using acoustic emission and optical coherence tomography . OBJECTIVE : CAD/ P62158 ceramic restorative material is routinely bonded to tooth substrates using adhesive cement . This study investigates micro-crack growth and damage in the ceramic/dentin adhesive interface under fatigue shear testing monitored using the acoustic emission ( AE ) technique with optical coherence tomography ( O75051 ) . METHODS : Ceramic/dentin adhesive samples were prepared to measure the shear bond strength ( SBS ) under static load . Fatigue shear testing was performed using a modified ISO14801 method . Loads in the fatigue tests were applied at 80 % , 70 % , and 60 % of the SBS to monitor interface debonding . The AE technique was used to detect micro-crack signals in static and fatigue shear bond tests . RESULTS : The results showed that the average SBS value in the static tests was 10.61±2.23MPa ( mean±standard deviation ) . The average number of fatigue cycles in which ceramic/dentin interface damage was detected in 80 % , 70 % and 60 % of the SBS were 152 , 1962 and 9646 , respectively . The acoustic behavior varied according to the applied load level . Events were emitted during 60 % and 70 % fatigue tests . A good correlation was observed between crack location in O75051 images and the number of AE signal hits . SIGNIFICANCE : The AE technique and O75051 images employed in this study could potentially be used as a pre-clinical assessment tool to determine the integrity of cemented load bearing restored ceramic material . Sustainable cyclic load stresses in ceramic/dentin-bonded specimens were substantially lower than the measured SBS . Predicted S-N curve showed that the maximum endured load was 4.18MPa passing 10(6) fatigue cyclic . A requirement for breast-cancer-associated gene 1 ( P38398 ) in the spindle checkpoint . P38398 -associated breast cancer exhibits significantly higher levels of chromosomal abnormalities than sporadic breast cancers . However , the molecular mechanisms regarding the roles of P38398 in maintaining genome integrity remain elusive . By using a mouse model deficient for Brca1 full-length isoform ( Brca1(Delta11/Delta11) ) , we found that Brca1(Delta11/Delta11) cells displayed decreased expression of a number of genes that are involved in the spindle checkpoint , including Mad2 , which is a key component of spindle checkpoint that inhibits anaphase-promoting complex . We showed that Brca1(Delta11/Delta11) cells failed to arrest at metaphase in the presence of DB08313 and underwent apoptosis because of activation of p53 . Consistently , reconstitution of Mad2 in Brca1(Delta11/Delta11) cells partially restored the spindle checkpoint and attenuated apoptosis . By using UBR60 cells , which carry tetracycline-regulated expression of P38398 , we demonstrated that P38398 binds to transcription factor O75051 -1 and up-regulates the transcription of Q13257 . Furthermore , we showed that the induction of P38398 to endogenous Q13257 or transfected Q13257 luciferase reporter in UBR60 cells was completely inhibited by acute suppression of P38398 by RNA interference . These data reveal a role of P38398 in maintaining genome integrity by interplaying with p53 and genes that are involved in the spindle checkpoint and apoptosis . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment . Altered growth factor expression in the aging penis : the Brown-Norway rat model . The objective of the present study was to evaluate age-related changes in the protein and gene expression of modulators of erectile function ( nitric oxide [ NO ] and endothelin-1 [ ET-1 ] ) and growth factors such as transforming growth factor ( TGF-beta1 ) and vascular endothelial growth factor ( P15692 ) in the penile tissue of Brown-Norway ( BN ) rats . Young and old BN male rats were euthanized , and the penile tissue was processed for immunohistochemical and molecular analyses . Total RNA was extracted , and an Access reverse transcription-polymerase chain reaction ( RT-PCR ) system was used for messenger RNA ( mRNA ) expression analysis . Immunohistochemical studies showed a decreased expression of endothelial nitric oxide synthase ( P29474 ) protein and an increased staining for ET-1 . Quantitative analysis of PCR products revealed decreased levels of P15692 mRNA expression in the old population of rats . The most significant decrease was detected between bands corresponding to splice forms 164 ( 21 % ) and 120 ( 18 % ) . The observed alterations in the gene expression of growth factors such as P15692 may contribute to the abnormal age-related morphological and physiological alterations in the erectile tissue . DB02901 interacts with P00533 /MAPK signalling and modulates P00533 levels in androgen receptor-positive LNCaP prostate cancer cells . P10275 ( AR ) signalling plays a pivotal role in prostate cancer pathogenesis and progression . However , androgen-mediated AR signalling is yet to be fully understood . P00533 and Q96HU1 kinase signalling pathways play predominant roles in AR function . Therefore , we investigated the interaction of P00533 signalling and AR activity in AR-positive LNCaP cells . We found that 5alpha-dihydrotestosterone ( DB02901 ) and P01133 had a synergistic effect on AR activity as detected by a luciferase reporter system , although P01133 alone did not activate AR . Both P27361 /2 and p38 were involved in DB02901 and DB02901 / P01133 -induced AR activation as detected by specific MEK and p38 inhibitors . Furthermore , 24-h treatment of the cells with DB02901 resulted in ubiquitination and down-regulation of the P00533 . This effect could be inhibited by the anti-androgen flutamide , suggesting an androgen-dependent mechanism . On the other hand , DB02901 -treatment strongly increased AR levels in LNCaP cells . These data suggest a complex regulatory loop between activated AR and P00533 . In conclusion , activation of AR by both DB02901 and P01133 / DB02901 involves the Q96HU1 kinase pathway . Long-term activation of AR results in increase of AR levels , which through so far unknown regulatory mechanisms results in ubiquitination and degradation of the P00533 . Treatment of peripapillary choroidal neovascularization with intravitreal bevacizumab . PURPOSE : Peripapillary choroidal neovascularization ( CNV ) is an uncommon condition and often shows a growth tendency towards the fovea during spontaneous progression that threatens visual acuity . Treatment of peripapillary CNV is difficult . The authors report results of intravitreal bevacizumab therapy for peripapillary CNV . METHODS : Four patients with CNV located in the temporal or superior peripapillary area received intravitreal bevacizumab injections . Ophthalmologic examinations including O75051 were performed at baseline and at 6-week intervals . DB00693 angiography was performed at baseline and depending on clinical and O75051 findings . The mean follow-up was 34+/-20 ( 22-69 ) weeks . RESULTS : The patients received an average of 3.5+/-3.1 ( 1-8 ) injections . In all patients fluorescein angiography showed inactivation of peripapillary CNV . No further increase in size was observed in any of the patients . The O75051 showed a decrease of intraretinal and subretinal fluid . No intraocular or systemic side effects were observed . CONCLUSIONS : In this series of patients , intravitreal bevacizumab appears to be efficacious . A progression of peripapillary CNV could be prevented in all patients and the lesion was successfully inactivated . Anti- P15692 treatment with bevacizumab represents a promising therapy option for peripapillary CNV . Gambogic acid inhibits angiogenesis through suppressing vascular endothelial growth factor-induced tyrosine phosphorylation of P35968 /Flk-1 . Previous studies revealed that gambogic acid ( GA ) , the major active ingredient of gamboge , a brownish to orange resin exuded from Garcinia hanburryi tree in Southeast Asia , possessed significant anticancer activity both in vitro and in vivo . In this study , we explored the high antiangiogenic activities of GA for the first time . GA inhibits the P15692 -stimulated proliferation , migration and tube formation of human umbilical vein endothelial cells ( HUVECs ) as well as microvessel sprouting from rat aortic rings in vitro . Moreover , GA inhibits vessel growth in matrigel plugs and P62158 in vivo and transplanted tumor in mice . The results also indicated that GA decreases P15692 production of cultured tumor cells and inhibits P15692 -induced tyrosine phosphorylation of P35968 /Flk-1 . This inhibition of receptor phosphorylation is correlated with a significant decrease in P15692 -triggered phosphorylated forms of P29323 , AKT and p38 . Taken together , these findings strongly suggest that GA might be a structurally novel angiogenesis inhibitor . Altered expression of beta-catenin , P12830 , cycloxygenase-2 , and p53 protein by ovine intestinal adenocarcinoma cells . Around 1.6 % of sheep in New Zealand develop small-intestinal adenocarcinomas . These neoplasms typically develop widespread metastases . The common development of these neoplasms and their subsequent behavior suggests that sheep could be a useful animal model of human colonic cancer . However , for an animal model of human disease to be relevant , similar genetic mutations should be present . Genetic mutations within human colonic cancers frequently result in expression of cycloxygenase-2 ( P35354 ) , loss of membranous expression of beta-catenin and P12830 , and accumulation of p53 protein within the neoplastic cells . Immunohistochemistry was used to investigate the presence of these 4 proteins within 26 ovine intestinal adenocarcinomas . Loss of membranous beta-catenin reactivity was observed in 14 of 26 ovine intestinal adenocarcinomas ( 54 % ) . The loss of membranous beta-catenin reactivity was accompanied by cytoplasmic and nuclear reactivity in 2 neoplasms . Loss of P12830 was observed within 8 of 26 neoplasms ( 31 % ) . Neoplastic cell expression of P35354 was observed in 12 of 26 neoplasms ( 46 % ) , whereas cells within 3 of 26 neoplasms ( 11 % ) contained visible p53 protein . In conclusion , all 4 proteins that commonly have altered expression in human colonic cancers were also altered in a proportion of the ovine intestinal adenocarcinomas . These results provide additional evidence that sheep could be useful for the study of human colonic cancer . 22-Oxacalcitriol prevents progression of endothelial dysfunction through antioxidative effects in rats with type 2 diabetes and early-stage nephropathy . BACKGROUND : Vitamin D deficiency is associated with endothelial dysfunction in type 2 diabetes patients , but the effectiveness of vitamin D supplementation remains controversial . We assessed whether 22-oxacalcitriol ( O75051 ) could prevent endothelial dysfunction in type 2 diabetes mellitus ( DM ) rats . METHODS : DM rats with early-stage nephropathy were treated for 10 weeks with O75051 ( 0.2 μg/kg ) three times per week or by an implanted insulin pellet . Endothelial dysfunction was assessed by femoral flow-mediated dilation ( FMD ) . RESULTS : P01308 significantly improved FMD as blood glucose levels normalized . O75051 also improved FMD without hypercalcemia or hyperphosphatemia and without affecting blood glucose or blood pressure . In femoral arteries , O75051 significantly suppressed the elevated expression of O75935 (phox) , a nicotinamide adenine dinucleotide phosphate ( NADPH ) oxidase subunit , and improved the endothelial nitric oxide synthase ( P29474 ) dimer-to-monomer ratio . In cultured endothelial cells , O75051 significantly inhibited high-glucose ( HG ) -induced reactive oxygen species ( ROS ) production . Simultaneously , O75051 significantly suppressed HG-induced O75935 (phox) expression and improved P29474 uncoupling as was observed in the in vivo study . CONCLUSION : In DM rats , O75051 improved endothelial dysfunction , at least in part , by suppressing ROS generation through O75935 (phox) expression , which might contribute to improving P29474 uncoupling . Adaptive optics imaging of cone mosaic abnormalities in acute macular neuroretinopathy . BACKGROUND AND OBJECTIVE : To assess the cone photoreceptor mosaic in acute macular neuroretinopathy ( Q9BXJ7 ) using adaptive optics ( AO ) imaging . PATIENTS AND METHODS : Four patients with Q9BXJ7 were evaluated retrospectively by near-infrared reflectance ( IR ) confocal scanning laser ophthalmoscopy ( Q12791 ) , spectral-domain optical coherence tomography ( SD- O75051 ) , and a flood-illuminated retinal AO camera . Microperimetry was performed in one patient . RESULTS : The cone photoreceptor density was decreased at the level of the Q9BXJ7 lesions . The cone mosaic disruption appeared heterogeneous and more widespread than the lesion detected in the IR- Q12791 and SD- O75051 images . The areas of cone loss correlated with SD- O75051 and microperimetry . After resolution of the Q9BXJ7 lesion on IR- Q12791 , there was incomplete recovery of the cone photoreceptor mosaic . CONCLUSION : Cone photoreceptor damage and reconstitution were documented in vivo at the cellular level in Q9BXJ7 using AO imaging . AO imaging appeared more sensitive than combined IR- Q12791 and SD- O75051 to detect and follow photoreceptor damage in patients with Q9BXJ7 . Kinin-B2 receptor exerted neuroprotection after diisopropylfluorophosphate-induced neuronal damage . The kinin-B2 receptor ( B2BKR ) activated by its endogenous ligand bradykinin participates in various metabolic processes including the control of arterial pressure and inflammation . Recently , functions for this receptor in brain development and protection against glutamate-provoked excitotoxicity have been proposed . Here , we report neuroprotective properties for bradykinin against organophosphate poisoning using acute hippocampal slices as an in vitro model . Following slice perfusion for 10min with diisopropylfluorophosphate ( DB00677 ) to initiate the noxious stimulus , responses of pyramidal neurons upon an electric impulse were reduced to less than 30 % of control amplitudes . Effects on synaptic-elicited population spikes were reverted when preparations had been exposed to bradykinin 30min after challenging with DB00677 . Accordingly , bradykinin-induced population spike recovery was abolished by HOE-140 , a B2BKR antagonist . However , the kinin-B1 receptor ( B1BKR ) agonist Lys-des- DB00125 (9)-bradykinin , inducing the phosphorylation of mitogen-activated protein kinase ( MEK/MAPK ) and cell death , abolished bradykinin-mediated neuroprotection , an effect , which was reverted by the P29323 inhibitor PD98059 . In agreement with pivotal B1BKR functions in this process , antagonism of endogenous B1BKR activity alone was enough for restoring population spike activity . On the other hand pralidoxime , an oxime , reactivating acetylcholinesterase ( P22303 ) after organophosphate poisoning , induced population spike recovery after DB00677 exposure in the presence of bradykinin and Lys-des- DB00125 (9)-bradykinin . Lys-des- DB00125 (9)-bradykinin did not revert protection exerted by pralidoxime , however when instead bradykinin and Ly-des- DB00125 (9)-bradykinin were superfused together , recovery of population spikes diminished . These findings again confirm the neuroprotective feature of bradykinin , which is , diminished by its endogenous metabolites , stimulating the B1BKR , providing a novel understanding of the physiological roles of these receptors . Distinct signalling pathways of murine histamine H1- and H4-receptors expressed at comparable levels in HEK293 cells . DB11320 ( HA ) is recognized by its target cells via four G-protein-coupled receptors , referred to as histamine H1-receptor ( P35367 ) , P25021 , Q9Y5N1 , and Q9H3N8 . Both P35367 and Q9H3N8 exert pro-inflammatory functions . However , their signal transduction pathways have never been analyzed in a directly comparable manner side by side . Moreover , the analysis of pharmacological properties of the murine orthologs , representing the main targets of pre-clinical research , is very important . Therefore , we engineered recombinant HEK293 cells expressing either mouse (m) P35367 or mH4R at similar levels and analyzed HA-induced signalling in these cells . HA induced intracellular calcium mobilization via both mH1R and mH4R , with the mH1R being much more effective . Whereas DB02527 accumulation was potentiated via the mH1R , it was reduced via the mH4R . The regulation of both second messengers via the Q9H3N8 , but not the P35367 , was sensitive to pertussis toxin ( PTX ) . The mitogen-activated protein kinases ( MAPKs ) P29323 1/2 were massively activated downstream of both receptors and demonstrated a functional involvement in HA-induced P18146 gene expression . The p38 MAPK was moderately activated via both receptors as well , but was functionally involved in HA-induced P18146 gene expression only in Q9H3N8 -expressing cells . Surprisingly , in this system p38 MAPK activity reduced the HA-induced gene expression . In summary , using this system which allows a direct comparison of mH1R- and mH4R-induced signalling , qualitative and quantitative differences on the levels of second messenger generation and also in terms of p38 MAPK function became evident . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . P10275 gene mutations in androgen insensitivity syndrome cause distinct patterns of reduced activation of androgen-responsive promoter constructs . Assessment of quantitative impairment of reporter gene activation is an important strategy proving pathogenetic relevance of androgen receptor ( AR ) -gene mutations in androgen insensitivity syndrome ( AIS ) . We hypothesized the additional existence of mutation-specific patterns of reduced target gene activation . Four AR-gene mutations causing AIS , L712F , M780I , R855H , and V866M , respectively , were recreated in an AR-expression plasmid . Activation of three structurally different androgen-dependent promoters ( MMTV , (ARE)2TATA , and GRE- O75051 ) was measured in transfected CHO-cells in response to dihydrotestosterone ( DB02901 ) , testosterone , androstenedione and stanozolol ( S ) . V866M showed the lowest activity across all conditions . R855H exhibited strikingly high activation of MMTV in response to DB02901 . M780I showed markedly low activation of (ARE)2TATA by S. L712F demonstrated high activation of GRE- O75051 . In essence , each mutation was characterized in this model by a specific pattern of reduced reporter gene activation . Our AR crystal structure analyses showed that L712 and M780 may cause distinct alterations of AR-ligand- and AR-coregulator interaction interfaces supporting the experimental observations . Our data support the hypothesis that mutations of the AR-gene in AIS induce mutation-specific patterns of reduced promoter activation in vitro . Considering the diversity of natural androgen-regulated promoters , mutation-specific differences of androgen response patterns may be of relevance in vivo and consequently may influence the AIS-phenotype . Assessment of transactivation patterns in vitro may be an interesting concept to extend functional description of AR-gene mutations in AIS . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 -I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 -Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 -I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 -Is . Treatment for the toxicity of P22303 -Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 -Is exert their effects . By attenuating the effects of P22303 -Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 -I . DB04630 stimulates vascular smooth muscle cell proliferation via big mitogen-activated protein kinase 1 activation . The nongenomic effects of aldosterone have been implicated in the pathogenesis of various cardiovascular diseases . DB04630 -induced nongenomic effects are attributable in part to the activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) , a classical mitogen-activated protein ( Q96HU1 ) kinase . Q13164 ( Q13164 ) , a newly identified Q96HU1 kinase , has been shown to be involved in cell proliferation , differentiation , and survival . We examined whether aldosterone stimulates Q13164 -mediated proliferation of cultured rat aortic smooth muscle cells ( RASMCs ) . P08235 ( MR ) expression and localization were evaluated by Western blotting analysis and fluorolabeling methods . P27361 /2 and Q13164 activities were measured by Western blotting analysis with the respective phosphospecific antibodies . Cell proliferation was determined by Alamar Blue colorimetric assay . DB04630 ( 0.1 to 100 nmol/L ) dose-dependently activated Q13164 in RASMCs , with a peak at 30 minutes . To clarify whether aldosterone-induced Q13164 activation is an MR-mediated phenomenon , we examined the effect of eplerenone , a selective MR antagonist , on aldosterone-induced Q13164 activation . DB00700 ( 0.1 to 10 micromol/L ) dose-dependently inhibited aldosterone-induced Q13164 activation in RASMCs . DB04630 also stimulated RASMC proliferation , which was inhibited by eplerenone . DB04630 -mediated phenomena were concluded to be attributable to a nongenomic effect because cycloheximide failed to inhibit aldosterone-induced Q13164 activation . Transfection of dominant-negative Q96HU1 kinase/ P29323 kinase 5 ( Q13163 ) , which is an upstream regulator of Q13164 , partially inhibited aldosterone-induced RASMC proliferation , which was almost completely inhibited by MEK inhibitor PD98059 . In addition to the classical steroid activity , rapid nongenomic effects induced by aldosterone may represent an alternative etiology for vascular diseases such as hypertension . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . P01308 / P05019 signaling pathways enhances tumor cell invasion through bisecting GlcNAc N-glycans modulation . an interplay with P12830 . Changes in glycosylation are considered a hallmark of cancer , and one of the key targets of glycosylation modifications is P12830 . We and others have previously demonstrated that P12830 has a role in the regulation of bisecting GlcNAc N-glycans expression , remaining to be determined the P12830 -dependent signaling pathway involved in this N-glycans expression regulation . In this study , we analysed the impact of P12830 expression in the activation profile of receptor tyrosine kinases such as insulin receptor ( IR ) and P08069 ( IGF-IR ) . We demonstrated that exogenous P12830 expression inhibits IR , IGF-IR and P29323 1/2 phosphorylation . Stimulation with insulin and P05019 in MDA-MD-435 cancer cells overexpressing P12830 induces a decrease of bisecting GlcNAc N-glycans that was accompanied with alterations on P12830 cellular localization . Concomitantly , IR/IGF-IR signaling activation induced a mesenchymal-like phenotype of cancer cells together with an increased tumor cell invasion capability . Altogether , these results demonstrate an interplay between P12830 and IR/IGF-IR signaling as major networking players in the regulation of bisecting N-glycans expression , with important effects in the modulation of epithelial characteristics and tumor cell invasion . Here we provide new insights into the role that P01308 / P05019 signaling play during cancer progression through glycosylation modifications . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Role of cardiovascular aldosterone in hypertension . DB04630 plays an important role in the pathogenesis of cardiovascular disease . We have reported that aldosterone is synthesized in cardiovascular tissues and local aldosterone synthesis plays important roles for hypertension and cardiac hypertrophy . High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP . Plasma aldosterone concentrations and P06703 were decreased by high salt intake in SHRSP . DB04630 production , the expression of P19099 mRNA and angiotensin II receptor AT1R mRNA in blood vessels were significantly increased by high salt intake . These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the vascular tissue in SHRSP , which may contribute to the development of malignant hypertension in salt-loaded SHRSP . However , there are several reports of conflicting data . P08235 ( MR ) binding is tightly regulated by the enzyme 11beta-hydroxysteroid dehydrogenase type 2 ( 11beta-HSD2 ) which selectively metabolizes glucocorticoids to inactive metabolites , thus allowing for MR activation by aldosterone . We have reported that decreased 11beta-HSD2 in blood vessels in Dahl salt-sensitive ( DS ) rats , a model for salt-sensitive hypertension . Local aldosterone excess may play a significant role in the salt sensitivity and development of hypertension . High sodium intake decreased circulating rennin-angiotensin-aldosterone system and increased blood pressure and cardiac hypertrophy in DS rats , which were prevented by the treatment with a selective MR antagonist , eplerenone . DB00700 also improved endothelial nitric oxide synthase ( P29474 ) activity and P29474 mRNA expression in blood vessels in DS rats . These results further suggest that not only circulating aldosterone but also local aldosterone is of critical importance in the pathophysiology of cardiovascular disorders . Inhibitor of G protein-coupled receptor kinase 2 normalizes vascular endothelial function in type 2 diabetic mice by improving β-arrestin 2 translocation and ameliorating Akt/ P29474 signal dysfunction . In type 2 diabetes , although Akt/endothelial NO synthase ( P29474 ) activation is known to be negatively regulated by G protein-coupled receptor kinase 2 ( P25098 ) , it is unclear whether the P25098 inhibitor would have therapeutic effects . Here we examined the hypotensive effect of the P25098 inhibitor and its efficacy agonist both vascular ( aortic ) endothelial dysfunction ( focusing especially on the Akt/ P29474 pathway ) and glucose intolerance in two type 2 diabetic models ( ob/ob mice and nicotinamide+streptozotocin-induced diabetic mice ) . Mice were treated with a single injection of the P25098 inhibitor or vehicle , and the therapeutic effects were compared by examining vascular function and by Western blotting . The P25098 inhibitor lowered blood pressure in both diabetic models but not in their age-matched controls . The P25098 inhibitor significantly improved clonidine-induced relaxation only in diabetic ( ob/ob and DM ) mice , with accompanying attenuations of P25098 activity and translocation to the plasma membrane . These protective effects of the P25098 inhibitor may be attributable to the augmented Akt/ P29474 pathway activation ( as evidenced by increases in Akt phosphorylation at DB00133 (473) and at DB00156 (308) , and P29474 phosphorylation at DB00133 (1177) ) and to the prevention of the P25098 translocation and promotion of β-arrestin 2 translocation to the membrane under clonidine stimulation . Moreover , the P25098 inhibitor significantly improved the glucose intolerance seen in the ob/ob mice . Our work provides the first evidence that in diabetes , the P25098 inhibitor ameliorates vascular endothelial dysfunction via the Akt/ P29474 pathway by inhibiting P25098 activity and enhancing β-arrestin 2 translocation under clonidine stimulation , thereby contributing to a blood pressure-lowering effect . We propose that the P25098 inhibitor may be a promising therapeutic agent for cardiovascular complications in type 2 diabetes . KR-31372 inhibits P35968 /Flk-1 tyrosine phosphorylation via K+( DB00171 ) channel opening in its antiangiogenic effect . The aim of this study was to identify the signaling pathway of the antiangiogenesis by ( 2R,3R,4S ) -N-cyano-N- ( 6-nitro-3,4-dihydro-hydroxy-2-methyl-2-dimethoxymethyl 2H-1-benzopyran-4yl ) -N'-benzylguanidine ( KR-31372 ) . KR-31372 inhibited the in vitro basal tube formation using Matrigel-coated plate and in vivo neovascularizations in mice induced by Matrigel containing vascular endothelial growth factor ( P15692 (165) , 5 ng/ml ) . P15692 (165) markedly increased cell proliferation using 5-bromo-2'-deoxyuridine incorporation and chemotactic migration using transwell chamber in human umbilical vein endothelial cells , those of which were significantly suppressed by pretreatment with KR-31372 and levcromakalim concentration dependently . The suppression of all these variables were strongly antagonized by glibenclamide , DB00171 -sensitive K(+) channel blocker . KR-31372 ( 10(-6)-10(-4) M ) and levcromakalim ( 10(-5) M ) concentration-dependently suppressed the P15692 (165)-induced increases in P35968 /Flk-1 tyrosine phosphorylation as well as the extracellular signal-related kinase 1/2 ( P27361 /2 ) , p38 MAK and p125( Q05397 ) tyrosine phosphorylation . These variables were significantly antagonized by glibenclamide . In conclusion , KR-31372 significantly inhibited the P35968 /Flk-1 tyrosine phosphorylation-linked P27361 /2 , p38 MAPK and p125( Q05397 ) tyrosine phosphorylation via mediation of K(+)( DB00171 ) channel opening , thereby resulting in antiangiogenesis . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Pharmacogenetics and future strategies in treating hyperglycaemia in diabetes . This review focuses on current evidence for pharmacogenetics for the 3 commonly used drug classes in treating diabetes : metformin , sulphonylureas and thiazolidinediones . Currently , metformin pharmacogenetics is focussing on drug transport with the recent finding that variation in O75051 transporters might affect metformin response . An aetiological approach has identified monogenic patients with diabetes due to TCF1 mutations who are particularly sensitive to the hypoglycaemic effects of sulphonylureas , and Q14654 or Q09428 mutations in which sulphonylureas can be used in place of insulin treatment . In Type 2 diabetes sulphonylurea response has been shown to be associated with variants Q9NQB0 associated with type 2 diabetes risk . For thiazolidinediones , focus has been on PPARgamma variants although with no consistent result . Genome wide association studies offer great potential to unravel what genetic factors influence response and side effects of diabetes therapies . Large numbers of well phenotyped patients for response and side effect as well as similarly sized similarly phenotyped replication cohorts are required . Establishing such cohorts is a priority in diabetes pharmacogenetics research . Is transforming growth factor-β signaling activated in human hypertrophied prostate treated by 5-alpha reductase inhibitor ? BACKGROUND AND AIM : It is well known that androgen deprivation relates to penile fibrosis , so we hypothesize that long-term treatment with 5-alphareductase inhibitors ( 5ARIs ) may increase the risk of fibrosis of prostate . PATIENTS AND METHODS : Thirty-two BPH patients who underwent transurethral resection of the prostate were enrolled . The patients were divided into two groups : group one , 16 patients underwent TURP who had been treated with tamsulosin for 2 years ; group two , 16 patients underwent TURP who had been treated with combination of tamsulosin and dutasteride for at least 1 year . We evaluated the expressions of P29475 , P35228 , P29474 , TGF-β1 , TGF-β2 , phosphorylated- Q15796 /3 ( p- Q15796 /3 ) , P12830 , P19022 , and α-smooth muscle actin in the resected prostate tissues by western blotting , and the TGF-β concentration was determined by ELISA kit . RESULTS : The expressions of 3 isoforms of NOS were significantly increased in group 2 except of P29474 in lateral prostate , and the expressions of TGF-β1 , TGF-β2 , and p- Q15796 /3 increased about 2-fold compared with group 1 . In group 2 , the P12830 expression decreased while P19022 expression increased significantly . CONCLUSIONS : The overexpression of P29475 may contribute to prostate smooth muscle relaxation ; however , long-time treatment with 5 Q9Y4X5 increases the risk of fibrosis of prostate . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . P08235 antagonism in the treatment of chronic central serous chorioretinopathy : a pilot study . PURPOSE : Based on experimental data showing that central serous chorioretinopathy could result from overactivation of mineralocorticoid receptor pathway in choroid vessels , the authors studied eplerenone , a mineralocorticoid receptor antagonist , as a potential treatment for chronic central serous chorioretinopathy . METHODS : This nonrandomized pilot study included 13 patients with central serous chorioretinopathy of at least 4-month duration , treated with 25 mg/day of oral eplerenone for a week followed by 50 mg/day for 1 or 3 months . The primary outcome measure was the changes in central macular thickness recorded by optical coherence tomography , and the secondary outcomes included changes in foveal subretinal fluid ( P11831 ) measured by O75051 , in best-corrected visual acuity ( BCVA ) and the percentage of eyes achieving complete resolution of subretinal fluid during the treatment period . RESULTS : Central macular thickness decreased significantly from 352 ± 139 μm at baseline to 246 ± 113 μm and 189 ± 99 μm at 1 and 3 months under eplerenone treatment ( P < 0.05 and P < 0.01 , respectively ) . At 3 months , the subretinal fluid significantly decreased compared with baseline subretinal fluid ( P < 0.01 ) and best-corrected visual acuity significantly improved compared with baseline best-corrected visual acuity ( P < 0.001 ) . CONCLUSION : DB00700 treatment was associated with a significant reduction in central macular thickness , subretinal fluid level , and an improvement in visual acuity . Randomized controlled trials are needed to confirm these encouraging results . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . Atrial natriuretic peptide : a possible mediator involved in dexamethasone 's inhibition of cell proliferation in multiple myeloma . Atrial natriuretic peptide ( P01160 ) has been recognized for several decades for its role of regulating blood pressure . Recently , cumulating evidences show that P01160 plays an anticancer role in various solid tumors via blocking the kinase cascade of Ras- Q02750 /2- P27361 /2 with the result of inhibition of DNA synthesis . P01160 , as well as its receptors ( P16066 and P17342 ) has been identified present in the embryonic stem cell and a wide range of cancer cells . Various lymphoid organs , such as lymph nodes , have been detected the presence of P01160 . Multiple myeloma ( MM ) , though the therapies have evolved significantly , is still an incurable disease as B lymphocyte cell neoplasm . Dexamethasone is the cornerstone in treatment of MM via inactivation of Ras- Q02750 /2- P27361 /2 cascade reaction . Coincidently , dexamethasone can increase the expression of P01160 markedly . Nevertheless , the role of P01160 in MM is unclear . Based on these results above , we raise the hypothesis that P01160 is involved in mediating dexamethasone 's inhibition of proliferation in MM cells , which suggests that P01160 may be a potential agent to treat MM . Role of genetic polymorphisms of ion channels in the pathophysiology of coronary microvascular dysfunction and ischemic heart disease . Conventionally , ischemic heart disease ( IHD ) is equated with large vessel coronary disease . However , recent evidence has suggested a role of compromised microvascular regulation in the etiology of IHD . Because regulation of coronary blood flow likely involves activity of specific ion channels , and key factors involved in endothelium-dependent dilation , we proposed that genetic anomalies of ion channels or specific endothelial regulators may underlie coronary microvascular disease . We aimed to evaluate the clinical impact of single-nucleotide polymorphisms in genes encoding for ion channels expressed in the coronary vasculature and the possible correlation with IHD resulting from microvascular dysfunction . 242 consecutive patients who were candidates for coronary angiography were enrolled . A prospective , observational , single-center study was conducted , analyzing genetic polymorphisms relative to ( 1 ) NOS3 encoding for endothelial nitric oxide synthase ( P29474 ) ; ( 2 ) P16615 encoding for the Ca²⁺/H⁺-ATPase pump ( SERCA ) ; ( 3 ) Q14524 encoding for the voltage-dependent Na⁺ channel ( Nav1.5 ) ; ( 4 ) Q15842 and Q14654 encoding for the Kir6.1 and Kir6.2 subunits of K- DB00171 channels , respectively ; and ( 5 ) KCN5A encoding for the voltage-gated K⁺ channel ( Kv1.5 ) . No significant associations between clinical IHD manifestations and polymorphisms for SERCA , Kir6.1 , and Kv1.5 were observed ( p > 0.05 ) , whereas specific polymorphisms detected in P29474 , as well as in Kir6.2 and Nav1.5 were found to be correlated with IHD and microvascular dysfunction . Interestingly , genetic polymorphisms for ion channels seem to have an important clinical impact influencing the susceptibility for microvascular dysfunction and IHD , independent of the presence of classic cardiovascular risk factors . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Anti-inflammatory activity of Taraxacum officinale . Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases . The dried plant was extracted with 70 % ethanol to generate its ethanol extract ( TEE ) . For some experiments , ethyl acetate ( EA ) , n-butanol ( BuOH ) and aqueous ( Aq ) fractions were prepared in succession from TEE . TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl ( DPPH ) assay , a diminishing effect on intracellular reactive oxygen species ( ROS ) level , and an anti-angiogenic activity in the chicken chorioallantoic ( P62158 ) assay . In the carrageenan-induced air pouch model , TEE inhibited production of exudate , and significantly diminished nitric oxide ( NO ) and leukocyte levels in the exudate . It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice . Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) in lipopolysaccharide ( LPS ) -stimulated macrophages were also assessed . Among the fractions , the n-butanol fraction ( BuOH ) was identified to be most effective in the P62158 assay . Collectively , Taraxacum officinale contains anti-angiogenic , anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and P35354 expression and/or its antioxidative activity . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . Early and delayed castrations confer a similar survival advantage in TRAMP mice . The most appropriate time to introduce androgen deprivation therapy for prostate cancer remains controversial . Our aim was to evaluate the effects of early versus delayed surgical castration on prostate cancer progression and survival in the transgenic adenocarcinoma of the mouse prostate ( TRAMP ) model . TRAMP mice were randomly divided into three groups : the early castration group ( on which castration was performed at the age of 4 weeks ) , the delayed castration group ( on which castration was performed when abdominal tumours could be palpated ) , and the sham-castrated group . Mice were monitored daily throughout their lives until cancer-related death or the development of an obviously moribund appearance , at which time the individual mouse was killed . P10275 expression in prostate tumours was also evaluated . The results shows that the average lifespan in early castration , delayed castration and sham-castrated groups were 54.1 weeks , 59.9 weeks and Q04695 weeks , respectively . Both early castration and delayed castration conferred a statistically significant survival advantage when compared with the sham-castrated group ( P < 0.001 ) . However , the difference in lifespan between the early castration group and the delayed castration group was not statistically significant ( P=0.85 ) . The increase in lifespan in the TRAMP mice that received either early or delayed castration correlated with lower G/B value ( genitourinary tract weight/body weight ) at death than the sham-castrated mice . In conclusion , early and delayed castrations in TRAMP mice prolonged survival to a similar extent . This finding may provide a guide for clinical practice in prostate cancer therapy . Different transport properties between famotidine and cimetidine by human renal organic ion transporters ( SLC22A ) . P25021 antagonist famotidine and cimetidine are commonly used for treatment of gastrointestinal ulcer diseases . Inasmuch as these drugs are mainly secreted by renal tubules , dosages have been adjusted according to renal function . Although many studies have been performed on the molecular mechanisms of renal handling of cimetidine , little is known about that of famotidine . In this study , to examine the recognition and transport of famotidine by human organic anion transporters ( OATs ; Q4U2R8 , Q8TCC7 ) and human organic cation transporter ( O75051 ; O15244 ) , the uptake studies using Xenopus laevis oocytes were performed in comparison with cimetidine . The half-maximal inhibitory concentrations of famotidine for [3H]estrone sulfate transport by Q8TCC7 and [14C]tetraethylammonium transport by O15244 ( 300 microM and 1.8 mM , respectively ) were higher than those of cimetidine ( 53 and 67 microM , respectively ) . While cimetidine inhibited p-[14C]aminohippurate transport by Q4U2R8 in a concentration dependent manner , famotidine did not affect it at 5 mM . In addition , Q8TCC7 mediated famotidine uptake , but Q4U2R8 and O15244 did not show famotidine transport . These results indicate that there are marked differences between famotidine and cimetidine in the recognition and transport by organic ion transporters and that Q8TCC7 contributes to the renal tubular secretion of famotidine . Present findings should be useful information to understand the renal handling of famotidine and cimetidine . Swept-source optical coherence tomography of lower limb wound healing with histopathological correlation . Direct noninvasive visualization of wound bed with depth information is important to understand the tissue repair . We correlate skin swept-source-optical coherence tomography ( O75051 ) with histopathological and immunohistochemical evaluation on traumatic lower limb wounds under honey dressing to compare and assess the tissue repair features acquired noninvasively and invasively . Analysis of optical biopsy identifies an uppermost brighter band for stratum corneum with region specific thickness ( p < 0.0001 ) and gray-level intensity ( p < 0.0001 ) variation . Below the stratum corneum , variation in optical intensities is remarkable in different regions of the wound bed . Correlation between O75051 and microscopic observations are explored especially in respect to progressive growth and maturation of the epithelial and subepithelial components . Characteristic transition of uniform hypolucid band in O75051 image for depigmented zone to wavy highly lucid band in the pigmented zone could be directly correlated with the microscopic findings . The transformation of prematured epithelium of depigmented area , with low expression of P12830 , to matured epithelium with higher P12830 expression in pigmented zone , implicated plausible change in their optical properties as depicted in O75051 . This correlated evaluation of multimodal images demonstrates applicability of swept-source- O75051 in wound research and importance of integrated approach in validation of new technology . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation .

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P46937 ( P46937 ) promotes human gallbladder tumor growth via activation of the P30530 /MAPK pathway . The transcriptional coactivator P46937 ( P46937 ) , a key regulator of cell proliferation and organ size in vertebrates , has been implicated in various malignancies . However , little is known about the expression and biological function of P46937 in human gallbladder cancer ( GBC ) . In this study we examined the clinical significance and biological functions of P46937 in GBC and found that nuclear P46937 and its target gene P30530 were overexpressed in GBC tissues . We also observed a significant correlation between high P46937 and P30530 expression levels and worse prognosis . The depletion of P46937 using lentivirus shRNAs significantly inhibited cell proliferation by inducing cell cycle arrest in S phase in concordance with the decrease of P24941 , P30304 , and cyclin A , and resulted in increased cell apoptosis and invasive repression in GBC cell lines in vitro . Furthermore , knockdown of P46937 also inhibited tumor growth in vivo . Additionally , we demonstrated that the activation of the P30530 /MAPK pathway was involved in the oncogenic functions of P46937 in GBC . These results demonstrated that P46937 is a putative oncogene and represents a prognostic marker and potentially a novel therapeutic target for GBC . Acetylbritannilactone suppresses lipopolysaccharide-induced vascular smooth muscle cell inflammatory response . To investigate the mechanism of action by which a new anti-inflammatory active compound , 1-O-acetylbritannilactone ( P00519 ) isolated from Inula britannica-F. , inhibits inflammatory responses in vascular smooth muscle cells ( VSMCs ) . Enzyme immunoassay was used to measure the levels of prostandin E(2) ( PGE(2) ) production . Immunocytochemistry staining and Western blot analysis were performed to detect the nuclear translocation of nuclear factor-kappaB ( NF-kappaB ) p65 and the expression of IkappaB-alpha , pIkappaB-alpha and cyclooxygenase-2 ( P35354 ) . Electrophoretic mobility shift assays ( EMSA ) were used to detect DNA-binding activity of NF-kappaB in VSMCs . P00519 ( 5 , 10 , 20 micrommol/l ) had several concentration-dependent effects , including inhibition of lipopolysaccharide ( LPS ) -induced PGE(2) production and P35354 expression , and blockade of NF-kappaB activation and translocation . These effects were owing to reductions in IkappaB-alpha phosphorylation and degradation induced by LPS . In addition , P00519 directly inhibited the binding of active NF-kappaB to specific DNA cis-element . These results indicate that P00519 is a potent inhibitor of LPS-stimulated VSMC inflammatory responses through blockade of NF-kappaB activity and inhibition of inflammatory gene P35354 expression . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . P01116 , P00533 , P09619 -α , P10721 and P35354 status in carcinoma showing thymus-like elements ( CASTLE ) . BACKGROUND : CASTLE ( Carcinoma showing thymus-like elements ) is a rare malignant neoplasm of the thyroid resembling lymphoepithelioma-like and squamous cell carcinoma of the thymus with different biological behaviour and a better prognosis than anaplastic carcinoma of the thyroid . METHODS : We retrospectively investigated 6 cases of this very rare neoplasm in order to investigate the mutational status of P01116 , P00533 , P09619 -α and P10721 , as well as the immunohistochemical expression pattern of CD117 , P00533 and P35354 , and possibly find new therapeutic targets . RESULTS : Diagnosis was confirmed by a moderate to strong expression of P06127 , CD117 and CK5/6 , whereas thyroglobulin , calcitonin and Q15669 -1 were negative in all cases . Tumors were also positive for P35354 and in nearly all cases for P00533 . In four cases single nucleotide polymorphisms ( SNPs ) could be detected in exon 12 of the P09619 -α gene ( rs1873778 ) , in three cases SNPs were found in exon 20 of the P00533 gene ( rs1050171 ) . No mutations were found in the P10721 and P01116 gene . CONCLUSIONS : All tumors showed a P35354 expression as well as an P00533 expression except for one case and a wild-type P01116 status . No activating mutations in the P00533 , P10721 and P09619 -α gene could be detected . Our data may indicate a potential for targeted therapies , but if these therapeutic strategies are of benefit in CASTLE remains to be determined . VIRTUAL SLIDES : The virtual slide(s) for this article can be found here : http://www.diagnosticpathology.diagnomx.eu/vs/1658499296115016 . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Cell cycle genes and ovarian cancer susceptibility : a tagSNP analysis . BACKGROUND : Dysregulation of the cell cycle is a hallmark of many cancers including ovarian cancer , a leading cause of gynaecologic cancer mortality worldwide . METHODS : We examined single nucleotide polymorphisms ( SNPs ) ( n=288 ) from 39 cell cycle regulation genes , including cyclins , cyclin-dependent kinases ( CDKs ) and CDK inhibitors , in a two-stage study . White , non-Hispanic cases ( n=829 ) and ovarian cancer-free controls ( n=941 ) were genotyped using an Illumina assay . RESULTS : Eleven variants in nine genes ( P00519 , O95067 , P38936 , P30281 , Q14209 , P24941 , O00716 , P06493 , and P50613 ) were associated with risk of ovarian cancer in at least one genetic model . Seven SNPs were then assessed in four additional studies with 1689 cases and 3398 controls . Association between risk of ovarian cancer and P00519 rs2855192 found in the original population [ odds ratio , OR ( BB vs AA ) 2.81 ( 1.29-6.09 ) , P=0.01 ] was also observed in a replication population , and the association remained suggestive in the combined analysis [ OR ( BB vs AA ) 1.59 ( 1.08-2.34 ) , P=0.02 ] . No other SNP associations remained suggestive in the replication populations . CONCLUSION : P00519 has been implicated in multiple processes including cell division , cell adhesion and cellular stress response . These results suggest that characterization of the function of genetic variation in this gene in other ovarian cancer populations is warranted . Sensitivity of breast cancer cells to erlotinib depends on cyclin-dependent kinase 2 activity . Inhibitors of epidermal growth factor receptor ( P00533 ) tyrosine kinases , such as erlotinib and gefitinib , have not been very effective in the treatment of breast cancer although many breast cancer cells express P00533 . To address this apparent paradox , we examined possible predictors of the sensitivity of 10 breast cancer cell lines to erlotinib in light of cyclin-dependent kinase 2 ( P24941 ) , considered the farthest downstream kinase that controls cell cycling in the P00533 signaling pathway . Expression of P00533 and P04626 were not associated with sensitivity to erlotinib . Expression of phosphorylated (p-)tyrosine , p-Akt , phosphorylated extracellular signal-regulated kinase ( p- P29323 ) 1/ P28482 ( Q8NFH3 / Q8TCB0 ) , and p27 after treatment of erlotinib was not associated with erlotinib sensitivity . However , suppression of P24941 activity after erlotinib treatment correlated with erlotinib sensitivity ( P < 0.0001 ) . Restoration of P24941 activity partially restored proliferation and induced erlotinib resistance in erlotinib-sensitive cell lines , indicating that sensitivity to erlotinib in these breast cancer cells depends , at least in part , on P24941 activity . p27 , an inhibitor of P24941 , was not translocated into the nucleus in erlotinib-resistant cell lines . Knocking down p27 protein partially blocked erlotinib-induced cell death and cell cycle arrest . These findings indicate that the ability of erlotinib to suppress P24941 activity is critical for cellular sensitivity to erlotinib , regardless of P00533 expression level , and that the presence of p27 in the cytoplasm also participates in erlotinib resistance . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . DB00184 activates P46937 through nAChRs mediated signaling in esophageal squamous cell cancer ( ESCC ) . Cigarette smoking is an established risk factor for esophageal cancers . P46937 ( P46937 ) , the key transcription factor of the mammalian Hippo pathway , has been reported to be an oncogenic factor for many cancers . In this study , we find nicotine administration can induce nuclear translocation and activation of P46937 in ESCC . Consistently , we observed nuclear translocation and activation of P46937 by knockdown of P32297 , which is a negative regulator of nicotine signaling in bronchial and esophageal cancer cells . DB00184 administration or P32297 depletion substantially increased proliferation and migration in esophageal cancer cells . Interestingly , we find that P46937 physically interacts with nAChRs , and nAChRs-signaling dissociates P46937 from its negative regulatory complex composed with α-catenin , β-catenin and 14-3-3 in the cytoplasm , leading to upregulation and nuclear translocation of P46937 . This process likely requires PKC activation , as PKC specific inhibitor Enzastaurin can block nicotine induced P46937 activation . In addition , we find nicotine signaling also inhibits the interaction of P46937 with Q9H3D4 , which contributes to the inhibitory effect of nicotine on apoptosis . Using immunohistochemistry analysis we observed upregulation of P46937 in a significant portion of esophageal cancer samples . Consistently , we have found a significant association between P46937 upregulation and cigarette smoking in the clinical esophageal cancer samples . Together , these findings suggest that the nicotine activated nAChRs signaling pathway which further activates P46937 plays an important role in the development of esophageal cancer , and this mechanism may be of a general significance for the carcinogenesis of smoking related cancers . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Expression of retinoic acid receptor beta in dermatofibrosarcoma protuberans . BACKGROUND : P10826 ( RAR beta ) has been shown to act as a tumor suppressor in many solid human tumors . To investigate the putative role of RAR beta in dermatofibrosarcoma protuberans ( DFSP ) , we examined the expression of RAR beta in DFSPs and analyzed the correlation of expression patterns between RAR beta and cyclooxygenase ( P36551 ) -2 as well as clinicopathological variables . METHODS : Using tissue microarray and immunohistochemistry , we evaluated nuclear RAR beta staining and cytoplasm P35354 staining in 53 DFSPs . RESULTS : 48 DFSPs ( 90.58 % ) were immunopositive for RAR beta , while 32 DFSPs ( 60.38 % ) were immunopositive for P35354 . RAR beta staining was significantly inversely correlated with P35354 staining ( p < 0.001 ; r =-0.668 ) . CONCLUSIONS : Our data indicated that RAR beta expressed in DFSPs and correlated with P35354 expression . RAR beta may be a potential therapeutic target for unresectable DFSP cases . Altered expression of beta-catenin , P12830 , cycloxygenase-2 , and p53 protein by ovine intestinal adenocarcinoma cells . Around 1.6 % of sheep in New Zealand develop small-intestinal adenocarcinomas . These neoplasms typically develop widespread metastases . The common development of these neoplasms and their subsequent behavior suggests that sheep could be a useful animal model of human colonic cancer . However , for an animal model of human disease to be relevant , similar genetic mutations should be present . Genetic mutations within human colonic cancers frequently result in expression of cycloxygenase-2 ( P35354 ) , loss of membranous expression of beta-catenin and P12830 , and accumulation of p53 protein within the neoplastic cells . Immunohistochemistry was used to investigate the presence of these 4 proteins within 26 ovine intestinal adenocarcinomas . Loss of membranous beta-catenin reactivity was observed in 14 of 26 ovine intestinal adenocarcinomas ( 54 % ) . The loss of membranous beta-catenin reactivity was accompanied by cytoplasmic and nuclear reactivity in 2 neoplasms . Loss of P12830 was observed within 8 of 26 neoplasms ( 31 % ) . Neoplastic cell expression of P35354 was observed in 12 of 26 neoplasms ( 46 % ) , whereas cells within 3 of 26 neoplasms ( 11 % ) contained visible p53 protein . In conclusion , all 4 proteins that commonly have altered expression in human colonic cancers were also altered in a proportion of the ovine intestinal adenocarcinomas . These results provide additional evidence that sheep could be useful for the study of human colonic cancer . Novel compound 1,3-bis ( 3,5-dichlorophenyl ) urea inhibits lung cancer progression . The successful clinical management of lung cancer is limited by frequent loss-of-function mutations in p53 which cooperates with chronic oxidant-stress induced adaptations in mercapturic acid pathway ( Q96HU1 ) which in turn regulates critical intracellular signaling cascades that determine therapeutic refractoriness . Hence , we investigated the anti-cancer effects and mechanisms of action of a novel compound called 1,3-bis(3,5-dichlorophenyl) urea ( COH-SR4 ) in lung cancer . Treatment with COH-SR4 effectively inhibited the survival and clonogenic potential along with inducing apoptosis in lung cancer cells . COH-SR4 treatment caused the inhibition of Q86UG4 activity and G0/ P55008 cell cycle arrest and inhibited the expression of cell cycle regulatory proteins P24941 , P11802 , cyclin A , cyclin B1 , cyclin E1 , and p27 . The COH-SR4 activated AMPK pathway and knock-down of AMPK partially reversed the cytotoxic effects of COH-SR4 in lung cancer . COH-SR4 treatment lead to regression of established xenografts of H358 lung cancer cells without any overt toxicity . The histopathology of resected tumor sections revealed an increase in pAMPK , a decrease in the nuclear proliferative marker Ki67 and angiogenesis marker CD31 . Western-blot analyses of resected tumor lysates revealed a decrease in pAkt and anti-apoptotic protein Bcl2 along with an increase in pAMPK , pro-apoptotic protein Bax and cleaved PARP levels . Importantly , COH-SR4 lead to decrease in the mesenchymal marker vimentin and increase in the normal epithelial marker P12830 . The results from our in-vitro and in-vivo studies reveal that COH-SR4 represents a novel candidate with strong mechanistic relevance to target aggressive and drug-resistant lung tumors . Transgenic Panax ginseng inhibits the production of P01375 , P05231 , and P10145 as well as P35354 expression in human mast cells . The most well-known medicinal plant , Panax ginseng ( P. ginseng ) , contains various phytosterols and bioactive triterpene saponins ( ginsenosides ) . P37268 is a key regulatory enzyme for triterpene biosynthesis and overexpression of the squalene synthase confers the hyper-production of triterpene saponins to form transgenic ginseng . In this study , we have investigated whether and how transgenic P. ginseng modulates an inflammatory reaction in a stimulated human mast cell line , HMC-1 . It was found that transgenic P. ginseng inhibited the production of tumor necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -6 , P10145 , and the expression of cyclooxygenase-2 in phorbol 12-myristate 13-acetate ( PMA ) plus calcium ionophore A23187 ( PMACI ) -stimulated HMC-1 . Additionally , we have shown that transgenic P. ginseng suppressed the intracellular calcium level induced by PMACI . These results provide new insights into the pharmacological actions of transgenic P. ginseng as a potential molecule for use in therapy in mast cell-mediated inflammatory diseases . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used . Quantum mechanics-based properties for 3D-QSAR . We have used a set of four local properties based on semiempirical molecular orbital calculations ( electron density ( ρ ) , hydrogen bond donor field ( HDF ) , hydrogen bond acceptor field ( P00748 ) , and molecular lipophilicity potential ( MLP ) ) for 3D-QSAR studies to overcome the limitations of the current force field-based molecular interaction fields ( MIFs ) . These properties can be calculated rapidly and are thus amenable to high-throughput industrial applications . Their statistical performance was compared with that of conventional 3D-QSAR approaches using nine data sets ( angiotensin converting enzyme inhibitors ( P12821 ) , acetylcholinesterase inhibitors ( AchE ) , benzodiazepine receptor ligands ( BZR ) , cyclooxygenase-2 inhibitors ( P35354 ) , dihydrofolate reductase inhibitors ( P00374 ) , glycogen phosphorylase b inhibitors ( GPB ) , thermolysin inhibitors ( THER ) , thrombin inhibitors ( THR ) , and serine protease factor Xa inhibitors ( fXa ) ) . The 3D-QSAR models generated were tested thoroughly for robustness and predictive ability . The average performance of the quantum mechanical molecular interaction field ( QM-MIF ) models for the nine data sets is better than that of the conventional force field-based MIFs . In the individual data sets , the QM-MIF models always perform better than , or as well as , the conventional approaches . It is particularly encouraging that the relative performance of the QM-MIF models improves in the external validation . In addition , the models generated showed statistical stability with respect to model building procedure variations such as grid spacing size and grid orientation . QM-MIF contour maps reproduce the features important for ligand binding for the example data set ( factor Xa inhibitors ) , demonstrating the intuitive chemical interpretability of QM-MIFs . P00797 angiotensin system modulates P42345 pathway through AT2R in HIVAN . P42345 ( P42345 ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 and p70S6K . DB09026 , a renin inhibitor attenuated phosphorylation of both P42345 and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 ) phosphorylation of p70S6K in a dose dependent manner . HIV/ P04629 also displayed enhanced phosphorylation of both P42345 and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 and p70S6K . These findings indicate that HIV stimulates P42345 pathway in HIVAN through the activation of renin angiotensin system via AT2R . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . P11511 inhibitors and cyclooxygenase-2 ( P35354 ) inhibitors in endometriosis : new questions -- old answers ? The medical treatment of endometriosis needs to be optimized . Therapeutic management strategies for endometriosis-associated pain or recurrent disease are primarily aimed at downregulating ovarian function or antagonizing the effect of estrogen in ectopic endometrial implants . In this context , basic research is providing important results for the development of new , specific treatment modalities . P11511 overexpression has recently been detected in endometriotic tissue . P11511 ( p450arom ) is responsible for converting C19 androgens into estrogen in several types of human tissue . P11511 activity causes local estrogen biosynthesis , which , in turn , stimulates prostaglandin E2 production by upregulating cyclooxygenase-2 ( P35354 ) . Thus , a positive feedback cycle develops between the two systems . Another abnormality in endometriosis , the deficient 17beta-hydroxysteroiddehydrogenase type II ( 17beta-HSD-Type-II ) expression , impairs the inactivation of estradiol to estrone . In contrast to the eutopic endometrium , these molecular aberrations increase the amount of local estradiol and prostaglandin E2 in endometriosis . In several human cell lines , prostaglandin and estrogen concentrations are associated with proliferation , migration , angiogenesis , apoptosis resistance and even invasiveness . Consequently , aromatase and P35354 are thought to be promising new therapeutic targets . Thus , specific aromatase inhibitors ( e.g. DB01006 / DB01006 , DB01217 /Arimidex or Exemestan/Aromasin ) or selective P35354 inhibitors ( e.g. Celecoxib/ DB00482 , DB00533 /Vioxx , DB00580 /Bextra ) are of great interest and should be studied in clinical trials in premenopausal woman with endometriosis to expand the spectrum of currently available treatment options . Epileptogenesis and reduced inward rectifier potassium current in tuberous sclerosis complex-1-deficient astrocytes . PURPOSE : Individuals with tuberous sclerosis complex ( TSC ) frequently have intractable epilepsy . To gain insights into mechanisms of epileptogenesis in TSC , we previously developed a mouse model of TSC with conditional inactivation of the Tsc1 gene in glia ( Tsc1( P14136 )CKO mice ) . These mice develop progressive seizures , suggesting that glial dysfunction may be involved in epileptogenesis in TSC . Here , we investigated the hypothesis that impairment of potassium uptake through astrocyte inward rectifier potassium ( Kir ) channels may contribute to epileptogenesis in Tsc1( P14136 )CKO mice . METHODS : Kir channel function and expression were examined in cultured Tsc1-deficient astrocytes . Kir mRNA expression was analyzed in astrocytes microdissected from neocortical sections of Tsc1( P14136 )CKO mice . Physiological assays of astrocyte Kir currents and susceptibility to epileptiform activity induced by increased extracellular potassium were further studied in situ in hippocampal slices . RESULTS : Cultured Tsc1-deficient astrocytes exhibited reduced Kir currents and decreased expression of specific Kir channel protein subunits , Kir2.1 and Kir6.1 . mRNA expression of the same Kir subunits also was reduced in astrocytes from neocortex of Tsc1( P14136 )CKO mice . By using pharmacologic modulators of signalling pathways implicated in TSC , we showed that the impairment in Kir channel function was not affected by rapamycin inhibition of the P42345 /S6K pathway , but was reversed by decreasing P24941 activity with roscovitine or retinoic acid . Last , hippocampal slices from Tsc1( P14136 )CKO mice exhibited decreased astrocytic Kir currents , as well as increased susceptibility to potassium-induced epileptiform activity . CONCLUSIONS : Impaired extracellular potassium uptake by astrocytes through Kir channels may contribute to neuronal hyperexcitability and epileptogenesis in a mouse model of TSC . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Toll-like receptor expression in human keratinocytes : nuclear factor kappaB controlled gene activation by Staphylococcus aureus is toll-like receptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent . Cultured primary human keratinocytes were screened for their expression of various members of the toll-like receptor ( TLR ) family . Keratinocytes were found to constitutively express Q15399 , O60603 , O15455 , O60602 , and Q9NR96 but not O00206 , Q9Y2C9 , Q9NYK1 , Q9NR97 , or Q9BXR5 as shown by polymerase chain reaction analysis . The expression of the crucial receptor for signaling of staphylococcal compounds O60603 was also confirmed by immunohistochemistry , in contrast to O00206 , which showed a negative staining pattern . Next , we analyzed the activation of the proinflammatory nuclear transcription factor kappaB by Staphylococcus aureus strain 8325-4 . Using nuclear extract gel shifts , RelA staining , and luciferase reporter transfection plasmids we found a clear induction of nuclear factor kappaB translocation by the bacteria . This translocation induced the transcription of nuclear factor kappaB controlled genes such as inducible nitric oxide synthetase , P35354 , and interleukin-8 . Transcription of these genes was followed by production of increased amounts of interleukin-8 protein and NO . Inhibition experiments using monoclonal antibodies and the specific platelet activating factor receptor inhibitor CV3988 showed that nuclear factor kappaB activation by S. aureus was O60603 but not O00206 or platelet activating factor receptor dependent . In line , the purified staphylococcal cell wall components lipoteichoic acid and peptidoglycan , known to signal through O60603 , also showed nuclear factor kappaB translocation in human keratinocytes , indicating a crucial role of the staphylococcal cell wall in the innate immune stimulation of human keratinocytes . These results help to explain the complex activation of human keratinocytes by S. aureus and its cell wall components in various inflammatory disorders of the skin . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Potential antitumor mechanisms of phenothiazine drugs . In this study , three kinds of phenothiazine drugs were analyzed to explore their potential antitumor mechanisms . First , target proteins that could interact with chlorpromazine , fluphenazine and trifluoperazine were predicted . Then , the target proteins of the three drugs were intersected . Cell signaling pathway enrichment and related disease enrichment were conducted for the intersected proteins to extract the enrichment categories associated with tumors . By regulation network analysis of the protein interactions , the mechanisms of action of these target proteins in tumor tissue were clarified , thus confirming the potential antitumor mechanisms of the phenothiazine drugs . The final results of cell signaling pathway enrichment and related disease enrichment showed that the categories with the highest score were all found in tumors . Target proteins belonging to the tumor category included signaling pathway members such as Wnt , MAPK and retinoic acid receptor . Moreover , another target protein , P45983 , could indirectly act on target proteins P24941 , P08069 , P49841 , P10276 , P21802 and P53779 , thereby affecting tumor cell division and proliferation . Therefore , phenothiazine drugs may have potential antitumor effects , and tumor-associated target proteins play important roles in the process of cell signaling transduction cascades . Feature-map vectors : a new class of informative descriptors for computational drug discovery . In order to develop robust machine-learning or statistical models for predicting biological activity , descriptors that capture the essence of the protein-ligand interaction are required . In the absence of structural information from X-ray or NMR experiments , deriving informative descriptors can be difficult . We have developed feature-map vectors ( FMVs ) , a new class of descriptors based on chemical features , to address this challenge . FMVs , which are derived from the conformational models of a few actives , are low dimensional , problem specific , and highly interpretable . By using shape-based alignments and scoring with chemical features , FMVs can combine information about a molecule 's shape and the pharmacophores it can match . In five validation studies , bag classifiers built using FMVs have shown high enrichments for identifying actives for five diverse targets : P24941 , 5-HT(3) , P00374 , thrombin , and P12821 . The interpretability of these descriptors has been demonstrated for P24941 and 5-HT(3) , where the method automatically discovers the standard literature pharmacophore . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Coadministrating luteolin minimizes the side effects of the aromatase inhibitor letrozole . P11511 inhibitors ( AIs ) have been used as adjuvant therapeutic agents for breast cancer . Their adverse side effect on blood lipid is well documented . Some natural compounds have been shown to be potential AIs . In the present study , we compared the efficacy of the flavonoid luteolin to the clinically approved AI letrozole ( DB01006 ; Novartis Pharmaceuticals , East Hanover , NJ ) in a cell and a mouse model . In the in vitro experimental results for aromatase inhibition , the Ki values of luteolin and letrozole were estimated to be 2.44 µM and 0.41 nM , respectively . Both letrozole and luteolin appeared to be competitive inhibitors . Subsequently , an animal model was used for the comparison . P11511 -expressing MCF-7 cells were transplanted into ovariectomized athymic mice . Luteolin was given by mouth at 5 , 20 , and 50 mg/kg , whereas letrozole was administered by intravenous injection . Similar to letrozole , luteolin administration reduced plasma estrogen concentrations and suppressed the xenograft proliferation . The regulation of cell cycle and apoptotic proteins-such as a decrease in the expression of Bcl-xL , cyclin-A/D1/E , P24941 /4 , and increase in that of Bax-was about the same in both treatments . The most significant disparity was on blood lipids . In contrast to letrozole , luteolin increased fasting plasma high-density lipoprotein concentrations and produced a desirable blood lipid profile . These results suggested that the flavonoid could be a coadjuvant therapeutic agent without impairing the action of AIs . A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion .

MH_train_16

MH_train_16

MH_train_16

interacts_with DB01233?

multiple_choice

DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Characterization of a novel Q13639 receptor antagonist of the azabicycloalkyl benzimidazolone class : DAU 6285 . Three chemical classes of serotonin Q13639 receptor agonists have been identified so far : 5-substituted indoles ( e.g. 5-HT ) , benzamides ( e.g. renzapride ) and benzimidazolones ( e.g. BIMU 8 ) . In a search for Q13639 receptor antagonists , we have discovered that the benzimidazolone derivative DAU 6285 ( for structure see text ) , is 3-5 times more potent than tropisetron in blocking 5-HT , renzapride and BIMU 8 induced stimulation of adenylate cyclase activity in mouse embryo colliculi neurons . Schild plot analysis yielded Ki values of 220 , 181 and 255 nmol/l , respectively . In addition , DAU 6285 showed poor activity as a 5- Q9H205 receptor ligand with respect to tropisetron , as demonstrated by in vitro binding studies ( Ki , 322 vs 2.8 nmol/l ) and by its antagonistic activity in the Bezold-Jarisch reflex test ( ID50 , 231 vs 0.5 micrograms/kg , i.v. ) . No significant binding ( Ki greater than 10 mumol/l ) of DAU 6285 to serotonergic P08908 , P28222 , P28335 , P28221 , and 5-HT2 receptors as well as to adrenergic alpha 1 , alpha 2 , dopaminergic D1 , D2 or muscarinic M1-M3 receptor subtypes was found . The data indicate that DAU 6285 has a somewhat higher affinity than tropisetron for Q13639 receptors , a property confirmed in functional tests , and much lower affinity than tropisetron for 5- Q9H205 receptors . The compound represents a new interesting tool for investigating the pharmacological and physiological properties of Q13639 receptors . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Therapy with interferon-beta modulates endogenous catecholamines in lymphocytes of patients with multiple sclerosis . OBJECTIVE : To investigate the endogenous dopaminergic/adrenergic system of lymphocytes in multiple sclerosis ( MS ) patients during treatment with interferon ( IFN ) -beta . METHODS : Patients with relapsing-remitting MS undergoing IFN-beta treatment were prospectively studied during the first year of treatment . Circulating lymphocytes were obtained at baseline and after 1 , 3 , 6 and 12 months of treatment and assayed for catecholamine ( CA ) production and mRNA expression of tyrosine hydroxylase ( TH , the rate-limiting enzyme in the synthesis of CA ) , beta(2)-adrenoceptors ( AR ) and D2 , D3 and D5 dopaminergic receptors ( DR ) . RESULTS : In cells from patients treated with IFN-beta for 12 months the production of CA hugely increased and was less sensitive to P01579 -induced inhibition . Expression of mRNA for TH , beta(2)-AR and P21918 was already enhanced after 1 month and further increased up to 6-12 months of treatment . On the contrary , P14416 mRNA progressively decreased and P35462 mRNA did not significantly change over the whole study period . CONCLUSIONS : In MS patients IFN-beta treatment enhances the ability of lymphocytes to produce CA , and induces extensive modifications of both beta(2)-AR and DR-operated pathways . The clinical relevance of these effects deserves consideration . Identification of novel genetic alterations in samples of malignant glioma patients . Glioblastoma is the most frequent and malignant human brain tumor . High level of genomic instability detected in glioma cells implies that numerous genetic alterations accumulate during glioma pathogenesis . We investigated alterations in AP-PCR DNA profiles of 30 glioma patients , and detected specific changes in 11 genes not previously associated with this disease : Q86UP9 , Q13326 , Q13639 , P05556 , P31327 , P07225 , P55259 , Q9UJ96 , Q08499 , Q8N743 , and Q14642 . Further correlations revealed that 8 genes might play important role in pathogenesis of glial tumors , while changes in P55259 , Q9UJ96 and Q8N743 should be considered as passenger mutations , consequence of high level of genomic instability . Identified genes have a significant role in signal transduction or cell adhesion , which are important processes for cancer development and progression . According to our results , Q86UP9 might be characteristic of primary glioblastoma , Q13326 , Q13639 , P05556 , P31327 , P07225 and Q14642 were detected predominantly in anaplastic astrocytoma , suggesting their role in progression of secondary glioblastoma , while alterations of Q08499 seem to have important role in development of both glioblastoma subtypes . Some of the identified genes showed significant association with p53 , p16 , and P00533 , but there was no significant correlation between loss of P60484 and any of identified genes . In conclusion our study revealed genetic alterations that were not previously associated with glioma pathogenesis and could be potentially used as molecular markers of different glioblastoma subtypes . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Signatures of positive selection in genes associated with human skin pigmentation as revealed from analyses of single nucleotide polymorphisms . Phenotypic variation between human populations in skin pigmentation correlates with latitude at the continental level . A large number of hypotheses involving genetic adaptation have been proposed to explain human variation in skin colour , but only limited genetic evidence for positive selection has been presented . To shed light on the evolutionary genetic history of human variation in skin colour we inspected 118 genes associated with skin pigmentation in the Perlegen dataset , studying single nucleotide polymorphisms ( SNPs ) , and analyzed 55 genes in detail . We identified eight genes that are associated with the melanin pathway ( Q9UMX9 , Q04671 , P17643 , P40126 , P21583 , P00533 , P14416 and Q03181 ) and presented significant differences in genetic variation between Europeans , Africans and Asians . In six of these genes we detected , by means of the EHH test , variability patterns that are compatible with the hypothesis of local positive selection in Europeans ( Q04671 , P17643 and P21583 ) and in Asians ( Q04671 , P40126 , P21583 , P00533 and P14416 ) , whereas signals were scarce in Africans ( P40126 , P00533 and P14416 ) . Furthermore , a statistically significant correlation between genotypic variation in four pigmentation candidate genes and phenotypic variation of skin colour in 51 worldwide human populations was revealed . Overall , our data also suggest that light skin colour is the derived state and is of independent origin in Europeans and Asians , whereas dark skin color seems of unique origin , reflecting the ancestral state in humans . Effects of cholinoceptor and 5-hydroxytryptamine3 receptor antagonism on erythromycin-induced canine intestinal motility disruption and emesis . 1. Erythromycin administration is associated with gastrointestinal problems , disturbed gastrointestinal motility and emesis . This study in the dog investigates the underlying mechanisms . 2 . Intestinal myoelectrical activity and the occurrence and latency of emesis were recorded in eight conscious dogs . All drugs were administered intravenously . 3 . Erythromycin ( 7 mg kg-1 ) increased contractions of the proximal small intestine , and caused emesis in all fasted dogs and in 5 dogs after food . Atropine ( 50 mg kg-1 min-1 ) and hexamethonium ( 10 mg kg-1 h-1 ) partially inhibited the GI motility effects but did not significantly reduce emesis . 4 . DB01233 at a high dose ( 2 mg kg-1 h-1 ) reduced the incidence of emesis in the presence of increased intestinal motility , but a low dose ( 150 micrograms kg-1 h-1 ) was ineffective . 5 . A 5-hydroxytryptamine3 ( 5- Q9H205 ) receptor antagonist , MDL 72222 ( 1 mg kg-1 ) , reduced emesis when given alone and combined with metoclopramide ( low dose ) . The Q13639 receptor agonist BRL24924 ( DB04917 , 1 mg kg-1 ) had no effect on emesis either alone in combination with metoclopramide . 6 . In conclusion , erythromycin-induced GI motility disturbances and emesis are not causally related . Whereas the increase in intestinal smooth muscle activity is possibly cholinergically mediated , emesis occurs at least in part via a 5-hydroxytryptaminergic mechanism , but does not involve the dopamine system . 5-hydroxytryptamine and its receptors in systemic vascular walls . 5-hydroxytryptamine ( 5-HT ) in the bloodstream is largely contained in platelets and circulates throughout the entire vascular system . 5-HT released from activated platelets dramatically changes the function of vascular smooth muscle cells ( VSMCs ) and endothelial cells ( ECs ) . In VSMCs , 5-HT induces proliferation and migration via 5- Q13049 receptors . These effects are further enhanced by vasoactive substances such as thromboxane A2 and angiotensin II . 5- Q13049 receptor activation in VSMCs also causes both enhancement of prostaglandin I2 production by inducing cyclooxygenase-2 and reduction of nitric oxide ( NO ) by suppressing inducible NO synthase . Evidence showing that 5-HT in ECs plays a principal role in angiogenesis now exists . Stimulation of 5-HT1 and/or 5-HT2 receptors has been implicated in the angiogenic effect of 5-HT . The extracellular signal-regulated kinase and endothelial NO synthase ( P29474 ) activation-dependent pathways are involved in the mechanisms . Moreover , Q13639 receptors in ECs have been shown to also regulate angiogenesis . Recent reports show sarpogrelate , a selective antagonist of the 5- Q13049 receptor , indirectly enhances the function of P28222 receptors in ECs via inhibition of 5- Q13049 receptors in VSMCs or platelets . This indirect action of P28222 receptors in ECs may increase NO production derived from P29474 and a vasodilator response . Furthermore , sarpogrelate and other 5- Q13049 receptor antagonists have been shown to reduce the constitutive activity of 5- Q13049 receptors . It is believed that increasing evidence on the role of 5-HT receptors will contribute to the expansion of the clinical application of existing therapeutic drugs such as sarpogrelate , and to the development of new 5-HT receptor-related drugs for treating cardiovascular diseases . DB01296 sulfate suppresses the expressions of urokinase plasminogen activator and inhibitor and gelatinases during the early stage of osteoarthritis . BACKGROUND : DB01296 sulfate may have an ex vivo inhibitory effect on the plasminogen activator ( PA ) /plasmin system and gelatinases expression during the early development of osteoarthritis ( OA ) . METHODS : We compared the levels of urokinase-type PA ( u-PA ) , PA inhibitor-1 ( P05121 ) and gelatinases ( matrix metalloproteinase-2 and -9 [ P08253 and -9 ] ) in a series of chondral , meniscal , and synovial cultures of early OA after treatment with or without glucosamine sulfate . RESULTS : Gelatin zymography revealed that glucosamine sulfate could suppress P08253 secretion in chondral , meniscal and synovial cultures and also decrease P14780 production in synovial and meniscal cultures . ELISA data also showed the suppressive effects of glucosamine sulfate on u-PA and P05121 production in synovial cultures at 48 h . CONCLUSIONS : Our data suggest that one of the therapeutic effects of glucosamine sulfate is to down-regulate the expressions of u-PA , P05121 , P08253 and P14780 that underlie the destruction of articular cartilage in the early stage of OA , and therefore to delay the joint failure . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Nonlinkage of bipolar illness to tyrosine hydroxylase , tyrosinase , and D2 and D4 dopamine receptor genes on chromosome 11 . OBJECTIVE : Previous linkage and allelic association studies using DNA polymorphisms , cosegregation of cytogenetic abnormalities with psychiatric illness , and assignment of genes involved in neutotransmitter metabolism suggested that chromosome 11 may harbor a gene predisposing to bipolar illness . The authors examined linkage in the families of 14 probands with bipolar illness , with the candidate genes tyrosine hydroxylase ( TH ) , D4 dopamine receptor ( P21917 ) at 11p15 , tyrosinase ( P14679 ) at 11q14-q21 , and D2 dopamine receptor ( P14416 ) at 11q22-q23 , as well as with the c-Harvey-ras oncogene ( P01112 ) and insulin gene ( P01308 ) , both located at 11p15 , a region that previously showed linkage to bipolar illness . METHOD : The genetic data were analyzed with both lod score analysis ( parametric ) and affected-sib-pair analysis ( nonparametric ) ; both narrow and broad definitions of the clinical phenotype were used . Further influences of diagnostic uncertainties were accounted for by using diagnostic probability classes weighing the stability of each phenotype . RESULTS : Two-point linkage results excluded close linkage of bipolar illness to each candidate gene ; negative results were also obtained when the narrow definition of the clinical phenotype was used . Moreover , multipoint linkage analysis of P01112 and P01308 excluded the 11p15 region encompassing both P21917 and TH . In agreement with the negative linkage results , affected-sib-pair analysis did not show preferential sharing of marker alleles at any of the candidate genes . CONCLUSIONS : The negative results obtained under different genetic models exclude a frequent role for P21917 , TH , P14679 , and P14416 in the pathogenesis of bipolar illness . 5-HT₄ receptor stimulation leads to soluble AβPPα production through P14780 upregulation . Serotonin 4 ( Q13639 ) receptor signaling does not only have the physiological function of improving cognition , but might also be helpful in the therapy of Alzheimer 's disease ( AD ) through regulation of the production of soluble amyloid-β protein precursor alpha ( sAβPPα ) . To analyze the relationship between Q13639 receptor signaling and sAβPPα production , we stably transfected H4 cells with AβPP and Q13639 receptor ( H4/AβPP/ Q13639 cells ) . We found that 24-h incubation with the Q13639 receptor agonist RS-67333 upregulates matrix metalloproteinase-9 ( P14780 ) . Furthermore , P14780 overexpression enhanced sAβPPα levels , whereas knockdown with P14780 siRNA decreased sAβPPα levels . When RS-67333 was injected for 10 days in Tg2576 mice , a model of amyloid-β peptide ( Aβ ) deposition , there was an increase in hippocampal levels of sAβPPα , C-terminal fragment α , and P14780 , as well as a decrease in hippocampal senile plaque number and levels of the 40 amino acid peptide , Aβ40 . Taken all together , these experiments demonstrate that Q13639 receptor stimulation induces expression of P14780 which cleaves AβPP through α-secretase-like activity , leading to an increase of sAβPPα levels and a reduction of Aβ load . Effect of metoclopramide , ondansetron and granisetron on aldosterone secretion in man . The plasma aldosterone response following the administration of drugs with antagonist and agonist activity at Serotonin 3 and 4 ( 5- Q9H205 & 4 ) receptors has been examined in 9 healthy male volunteers receiving the following four treatments i.v. in a randomised , cross-over sequence : ondansetron 8 mg , granisetron 3 mg , metoclopramide 20 mg , and saline 20 ml . DB01233 significantly increased the mean plasma aldosterone level to 196 % of basal level at 5 min . It rose to 234 % at 15 min and remained at more than 185 % of basal level for the duration of the experiment . The response to ondansetron and granisetron did not differ significantly from placebo . If dopamine antagonism is discounted , the results suggest that metoclopramide-induced aldosterone secretion results from its agonist activity at Q13639 receptors , although slow neuronal depolarization via an unidentified receptor remains a possibility . Antagonism at the 5- Q9H205 receptor plays no role , as the selective antagonist , granisetron , did not elicit a significant response . It seems unlikely that the Q13639 receptor is the second , low affinity binding site of ondansetron , unless it had no agonist activity at this receptor . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Effects of metoclopramide and tropisetron on aldosterone secretion possibly due to agonism and antagonism at the Q13639 receptor . OBJECTIVE : Part of the prokinetic activity of metoclopramide can possibly be ascribed to agonist activity at Q13639 receptors . The 5- Q9H205 antagonist tropisetron is thought to act as an antagonist at Q13639 receptors . In the present study aldosterone secretion in response to the administration of these two drugs was explored to examine the role of the Q13639 receptor in aldosterone secretion . METHODS : Following a single-blind , random design , ten normal male volunteers received one of the following regimens on three occasions , with at least 2-week intervals : metoclopramide 10 mg i.v. ; tropisetron 5 mg by slow i.v.i. , or ; tropisetron by slow i.v.i. , followed by 10 mg metoclopramide i.v. RESULTS : In response to metoclopramide alone the mean plasma aldosterone level rose significantly to 149 % of basal level and remained significantly elevated for the next 20 min . With tropisetron alone , there was a significant 37.8 % drop at 60 min and the aldosterone levels remained low for the duration of the experiment . DB01233 reversed the decline mediated by tropisetron significantly at 30 and 90 min . DB04630 levels after the latter regimen also did not differ significantly from baseline at any time period . CONCLUSION : These results would suggest the existence of a tonic stimulatory influence of 5-HT via Q13639 receptors on aldosterone secretion , which could be augmented by metoclopramide and blocked by tropisetron . However , the effect of tropisetron per se should be interpreted with caution given the lack of a saline group . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Analysis of neurogenic contractions induced by ML-1035 and other benzamides in the guinea-pig non-stimulated isolated ileum . 4-Amino-5-chloro-substituted benzamides have been shown to increase gastric motility in-vivo and enhance field-stimulated and peristaltic contractions in-vitro . The present experiments examined the contractile response to a series of benzamides in the guinea-pig non-stimulated ileum . Four benzamides elicited contractions in the isolated ileum which were expressed as a percentage of the contraction induced by 1 microM acetylcholine ( % acetylcholine response = 12 +/- 2 , 19 +/- 3 , 26 +/- 2 , 51 +/- 3 , n = 13 , 8 , 17 , and 21 , with EC50 values of 0.85 , 1.8 , 5.7 , and 14.2 microM for cisapride , zacopride , metoclopramide , and ML-1035 ( 4-amino-5-chloro-2-((2-methylsulphinyl)-ethoxy)-N- ( 2-(diethylamino)-ethyl ) -benzamide hydrochloride ) , respectively ) . ML-1035 contractions were completely blocked by atropine and tetrodotoxin , while ganglionic blockade with hexamethonium was ineffective . DB01233 has been reported to sensitize postjunctional muscarinic receptors , however , ML-1035 did not enhance acetylcholine-induced contractions . Tropisetron ( ICS 205-930 , 1 microM ) , caused a parallel rightward shift in the concentration-response curve for both ML-1035 and zacopride ( EC50 = 14.2 +/- 1.3 and 1.8 +/- 0.8 microM in the absence , and 26 +/- 2.7 and 6.9 +/- 2.3 microM in the presence of tropisetron for ML-1035 and zacopride , respectively ) with apparent pKB values of 5.9 and 6.0 for the respective compounds . 5-Hydroxytryptaminergic receptor desensitization by 2-methyl-5-hydroxytryptamine ( 5- Q9H205 ) and 5-methoxytryptamine ( Q13639 ) , attenuated the response to ML-1035. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . Enhanced incentive motivation for sucrose-paired cues in adolescent rats : possible roles for dopamine and opioid systems . Vulnerability to the effects of drugs of abuse during adolescence may be related to altered incentive motivation , a process believed to be important in addiction . Incentive motivation can be seen when a neutral stimulus acquires motivational properties through repeated association with a primary reinforcer . We compared adolescent ( postnatal day ( P01160 ) 24-50 ) and adult ( > P01160 70 ) rats on a measure of incentive motivation : responding for a conditioned reinforcer ( CR ) . Rats learned to associate the delivery of 0.1 ml of 10 % sucrose with a conditioned stimulus ( CS ; light and tone ) ; 30 pairings per day were given over 14 days . Then , we measured responding on a lever delivering the CS ( now a CR ) after injections of amphetamine ( 0 , 0.25 or 0.5 mg/kg ) . We also examined responding for CR when the CS and sucrose were paired or unpaired during conditioning , and responding for the primary reinforcer ( 10 % sucrose ) in control experiments . Finally , we examined the effects of D(1) and P14416 antagonists ( P35240 39166 and eticlopride , respectively ) and an opioid receptor antagonist ( naltrexone ) on responding for a CR in adolescent rats . Adolescents but not adults acquired responding for a CR , but adolescents responded less than adults for the primary reinforcer . Responding for a CR depended upon the pairing of the CS and sucrose during conditioning . Both dopamine and opioid receptor antagonists reduced responding for the CR . Therefore , incentive motivation may be enhanced in adolescents compared with adults , and incentive motivation may be mediated in part by both dopamine and opioid systems . Chronic atrial fibrillation alters the functional properties of If in the human atrium . INTRODUCTION : Despite the evidence that the hyperpolarization-activated current ( If ) is highly modulated in human cardiomyopathies , no definite data exist in chronic atrial fibrillation ( cAF ) . We investigated the expression , function , and modulation of If in human cAF . METHODS AND RESULTS : Right atrial samples were obtained from sinus rhythm ( SR , n = 49 ) or cAF ( duration > 1 year , n = 31 ) patients undergoing corrective cardiac surgery . Among f-channel isoforms expressed in the human atrium ( O60741 , 2 and 4 ) , Q9Y3Q4 mRNA levels measured by RT-PCR were significantly reduced . However , protein expression was preserved in cAF compared to SR ( +85 % for Q9Y3Q4 ) ; concurrently , miR-1 expression was significantly reduced . In patch-clamped atrial myocytes , current-specific conductance ( gf ) was significantly increased in cAF at voltages around the threshold for If activation ( -60 to -80 mV ) ; accordingly , a 10-mV rightward shift of the activation curve occurred ( P < 0.01 ) . β-Adrenergic and Q13639 receptor stimulation exerted similar effects on If in cAF and SR cells , while the P01160 -mediated effect was significantly reduced ( P < 0.02 ) , suggesting downregulation of natriuretic peptide signaling . CONCLUSIONS : In human cAF modifications in transcriptional and posttranscriptional mechanisms of HCN channels occur , associated with a slight yet significant gain-of-function of If , which may contribute to enhanced atrial ectopy . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . DB01233 does not increase gastric muscle contractility in newborn rats . Feeding intolerance resulting from delayed gastric emptying is common in premature neonates . DB01233 ( MCP ) , the most frequently used prokinetic drug in neonates , enhances gastric muscle contractility through inhibition of dopamine receptors . Although its therapeutic benefit is established in adults , limited data are available to support its clinical use in infants . Hypothesizing that developmentally dependent differences are present , we comparatively evaluated the effect of MCP on fundus muscle contractility in newborn , juvenile , and adult rats . The muscle strips were either contracted with electrical field stimulation ( O43281 ) to induce cholinergic nerve-mediated acetylcholine release or carbachol , a cholinergic agonist acting directly on the muscarinic receptor . Although in adult rats MCP increased O43281 -induced contraction by 294 ± 122 % of control ( P < 0.01 ) , no significant effect was observed in newborn fundic muscle . MCP had no effect on the magnitude of the carbachol-induced and/or bethanechol-induced gastric muscle contraction at any age . In response to dopamine , an 80.7 ± 5.3 % relaxation of adult fundic muscle was observed , compared with only a 8.4 ± 8.7 % response in newborn tissue ( P < 0.01 ) . P14416 expression was scant in neonates and significantly increased in adult gastric tissue ( P < 0.01 ) . In conclusion , the lack of MCP effect on the newborn fundic muscle contraction potential relates to developmental differences in dopamine D2 receptor expression . To the extent that these novel data can be extrapolated to neonates , the therapeutic value of MCP as a prokinetic agent early in life requires further evaluation . Tissue dependent differences in G-protein coupled receptor kinases associated with Q13639 receptor desensitization in the rat gastro-intestinal tract . Desensitization of 5-HT(4) receptors is regulated by G-protein coupled receptor kinases ( GRKs ) . However , the specific GRK(s) that regulates the desensitization of 5-HT(4) receptors in the in vivo setting is unknown . We investigated the in situ expression of 5-HT(4) receptors and the GRKs in the rat gastrointestinal tract using immunohistochemistry and their interaction using coimmunoprecipitation . 5-HT(4) receptors were expressed in the tunica muscularis mucosae of the oesophagus , longitudinal muscle , myenteric plexus , circular muscle , submucosal plexus and muscularis mucosae of both the proximal and distal colon . P25098 was expressed in longitudinal muscle and occasionally in myenteric plexus whilst P34947 showed limited expression in the nerve endings of the myenteric plexus and submucosal plexus of the colon . P35626 was expressed in the tunica muscularis mucosae of the oesophagus , circular muscle , submucosal plexus and muscularis mucosae of the colon . P43250 was expressed in the tunica muscularis mucosae of the oesophagus , longitudinal muscle , circular muscle , and muscularis mucosae of the colon . Stimulation of tunica muscularis mucosae of the oesophagus and distal colon using the 5-HT(4) receptor agonist , tegaserod , followed by analysis of the 5-HT(4) receptor antibody immunoprecipitate , revealed the coimmunoprecipitation of P43250 with 5-HT(4) receptors in the tunica muscularis mucosae of oesophagus while P25098 and P43250 were coimmunoprecipitated with 5-HT(4) receptors in the distal colon . This study indicates that P43250 may be involved in the regulation of the desensitization of 5-HT(4) receptors in the rat oesophagus whilst P25098 and P43250 may be involved in regulation of the desensitization of 5-HT(4) receptors in the distal colon . The relationship between gastric motility and nausea : gastric prokinetic agents as treatments . Nausea is one of a cluster of symptoms described subjectively by patients with delayed gastric emptying . The mechanisms and treatments are unclear ( anti-emetic drugs are not fully effective against nausea ) . Can nausea be relieved by stimulating gastric emptying ? Physostigmine ( together with atropine ) has been shown experimentally to stimulate gastric motility , relieve nausea and restore normal gastric motility . Is this mimicked by gastric prokinetic drugs ? The answer is complicated by mixed pharmacology . DB01233 increases gastric motility by activating myenteric Q13639 receptors but also directly inhibits vomiting via D2 and 5- Q9H205 receptor antagonism ; relationships between increased gastric motility and relief from nausea are therefore unclear . Similarly , the D2 receptor antagonist domperidone has direct anti-emetic activity . Nevertheless , more selective Q13639 and motilin receptor agonists ( erythromycin , directly stimulating gastric motility ) inhibit vomiting in animals ; low doses of erythromycin can also relieve symptoms in patients with gastroparesis . Ghrelin stimulates gastric motility and appetite mostly via vagus-dependent pathways , and inhibits vomiting in animals . To date , ghrelin receptor activation has failed to consistently improve gastric emptying or symptoms in patients with gastroparesis . We conclude that nausea can be relieved by gastric prokinetic drugs , but more clinical studies are needed using drugs with selective activity . Other mechanisms ( e.g. ghrelin , vagal and central pathways , influencing a mechanistic continuum between appetite and nausea ) also require exploration . These and other issues will be further explored in a forthcoming special issue of the European Journal of Pharmacology , which focusses on mechanisms of nausea and vomiting . P11387 is a cofactor for c-Jun in the regulation of epidermal growth factor receptor expression and cancer cell proliferation . P11387 ( Topo I ) is a molecular target for the anticancer agent topotecan in the treatment of small cell lung cancer and ovarian carcinomas . However , the molecular mechanisms by which topotecan treatment inhibits cancer cell proliferation are unclear . We describe here the identification of Topo I as a novel endogenous interaction partner for transcription factor c-Jun . Reciprocal coimmunoprecipitation analysis showed that Topo I and c-Jun interact in transformed human cells in a manner that is dependent on JNK activity . c-Jun target gene epidermal growth factor receptor ( P00533 ) was identified as a novel gene whose expression was specifically inhibited by topotecan . Moreover , Topo I overexpression supported c-Jun-mediated reporter gene activation and both genetic and chemical inhibition of c-Jun converted cells resistant to topotecan-elicited P00533 downregulation . DB01030 -elicited suppression of proliferation was rescued by exogenously expressed P00533 . Furthermore , we demonstrate the cooperation of the JNK-c-Jun pathway , Topo I , and P00533 in the positive regulation of O75794 cell proliferation . Together , these results have identified transcriptional coactivator Topo I as a first endogenous cofactor for c-Jun in the regulation of cell proliferation . In addition , the results of the present study strongly suggest that inhibition of P00533 expression is a novel mechanism by which topotecan inhibits cell proliferation in cancer therapy . DB00989 improves hippocampal neurogenesis and depression-like behaviors via P08908 receptor stimulation in olfactory bulbectomized mice . DB00989 is a non-competitive inhibitor of both acetylcholinesterase ( P22303 ) and butylcholinesterase ( BuChE ) used to treat mild to moderate dementia in Alzheimer 's disease ( AD ) patients . Although rivastigmine reportedly ameliorates cognitive dysfunction in these patients , its ability to improve Behavioral and Psychological Symptoms of Dementia ( BPSD ) remains unclear . To determine whether rivastigmine treatment antagonizes depression-like behaviors , we chronically administered rivastigmine ( 0.1-1.0mg/kg ) to olfactory bulbectomized ( OBX ) mice once a day for 2weeks , starting 2weeks after bulbectomy . Chronic treatment at 0.3 or 1.0mg/kg dose dependently and significantly improved depression-like behaviors , as assessed by tail suspension ( Q16762 ) , forced swim ( P19883 ) , locomotion and novelty-suppressed feeding ( NSFT ) tests . Importantly , co-administration with WAY-100635 ( 1.0mg/kg ) , a P08908 receptor antagonist , but not ketanserin ( 1.0mg/kg , ) , a 5- Q13049 receptor antagonist , completely blocked rivastigmine-induced anti-depressive effects , suggesting that P08908 receptor stimulation mediates this activity . Consistent with this observation , rivastigmine treatment significantly rescued impaired neurogenesis observed in OBX mice in a P08908 receptor-dependent manner . Furthermore , enhanced protein kinase B ( Akt ) and extracellular signal-regulated kinase ( P29323 ) phosphorylation seen following rivastigmine treatment was closely associated with improved neurogenesis . These effects were blocked by WAY-100635 but not ketanserin treatment . Finally , we confirmed that P08908 but not 5- Q13049 receptor stimulation by specific agonists mimicked rivastigmine-induced anti-depression activity and promoted hippocampal neurogenesis . We conclude that , in addition to enhancing the cholinergic system , rivastigmine treatment restores normal function of the hippocampal serotonergic system , an activity that likely ameliorates depressive behaviors in AD patients . Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . A whole genome Bayesian scan for adaptive genetic divergence in West African cattle . BACKGROUND : The recent settlement of cattle in West Africa after several waves of migration from remote centres of domestication has imposed dramatic changes in their environmental conditions , in particular through exposure to new pathogens . West African cattle populations thus represent an appealing model to unravel the genome response to adaptation to tropical conditions . The purpose of this study was to identify footprints of adaptive selection at the whole genome level in a newly collected data set comprising 36,320 SNPs genotyped in 9 West African cattle populations . RESULTS : After a detailed analysis of population structure , we performed a scan for SNP differentiation via a previously proposed Bayesian procedure including extensions to improve the detection of loci under selection . Based on these results we identified 53 genomic regions and 42 strong candidate genes . Their physiological functions were mainly related to immune response ( MHC region which was found under strong balancing selection , P11912 , P61073 , P80370 , P48380 , Q9H3S1 , Q8IUC6 and P19474 ) , nervous system ( Q96NK8 , O95897 , MAGI1 , Q9H3S1 and Q13639 ) and skin and hair properties ( P24530 , TRSP1 and Q8IUC2 ) . CONCLUSION : The main possible underlying selective pressures may be related to climatic conditions but also to the host response to pathogens such as Trypanosoma(sp) . Overall , these results might open the way towards the identification of important variants involved in adaptation to tropical conditions and in particular to resistance to tropical infectious diseases . The P38936 codon 31*C- and P14416 codon 313*T-related genotypes/alleles , but not P18887 codon 399 , hOGG1 codon 326 , and P21728 -48 polymorphisms , are correlated with the presence of leiomyoma . OBJECTIVE : To investigate whether the gene polymorphisms for P38936 , X-ray repair cross-complementing group 1 ( P18887 ) , human 8-oxoguanine glycosylase 1 ( hOGG1 ) , and dopamine D1 and D2 receptors ( P21728 , -2 ) are associated with leiomyoma susceptibility . DESIGN : Prospective study . SETTING : Departments of gynecology and genetics in a medical center . PATIENT(S) : Women were divided into two groups : leiomyoma ( n = 120 ) and nonleiomyoma ( n = 112 ) . INTERVENTION(S) : The P38936 codon 31 , P18887 codon 399 , hOGG1 codon 326 , P21728 -48 , and P14416 codon 313 polymorphisms were genotyped by polymerase chain reaction with restriction enzyme digestions ( Blp I , MspI , Fnu4HI , Dde I , and NcoI , respectively ) . MAIN OUTCOME MEASURE(S) : Genotypes and allelic frequencies . RESULT(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyomas . The proportions of P38936 (*)CC/CA/AA and P14416 (*)CC/CT/TT in both groups were 27.5/68.3/4.2 % and 12.5/51.7/35.8 % ( leiomyoma ) ; and 14.3/51.8/33.9 % and 33.9/40.2/25.9 % ( nonleiomyoma ) . P18887 , hOGG1 , and P21728 were not correlated with the presence of leiomyomas . P18887 (*)GG/GA/AA , hOGG1(*)TT/TA/AA , and P21728 (*)GG/GA/AA were 54.2/37.5/8.3 % , 36.7/44.2/19.1 % , and 3.3/25.8/70.8 % ( leiomyoma ) ; and 48.2/47.3/4.5 % , 43.6/41/15.4 % , and 3.6/25/71.4 % ( nonleiomyoma ) . CONCLUSION(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyoma . P18887 , hOGG1 , and P21728 were not correlated with leiomyoma development . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . DB01233 stimulates catecholamine- and granin-derived peptide secretion from pheochromocytoma cells through activation of serotonin type 4 ( Q13639 ) receptors . The gastroprokinetic agent metoclopramide is known to stimulate catecholamine secretion from pheochromocytomas . The aim of the study was to investigate the mechanism of action of metoclopramide and expression of serotonin type 4 ( 5-HT(4) ) receptors in pheochromocytoma tissues . Tissue explants , obtained from 18 pheochromocytomas including the tumor removed from a 46-year-old female patient who experienced life-threatening hypertension crisis after metoclopramide administration and 17 additional pheochromocytomas ( 9 benign and 8 malignant ) were studied . Cultured pheochromocytoma cells derived from the patient who previously received metoclopramide were incubated with metoclopramide and various 5-HT(4) receptor ligands . In addition , total mRNAs were extracted from all the 18 tumors . Catecholamine- and granin-derived peptide concentrations were measured in pheochromocytoma cell incubation medium by HPLC and radioimmunological assays . In addition , expression of 5-HT(4) receptor mRNAs in the 18 pheochromocytomas was investigated by the use of reverse transcriptase-PCR . RESULTS : DB01233 and the 5-HT(4) receptor agonist cisapride were found to activate catecholamine- and granin-derived peptide secretions by cultured tumor cells . DB01233 - and cisapride-evoked catecholamine- and granin-derived peptide productions were inhibited by the 5-HT(4) receptor antagonist GR 113808 . 5-HT(4) receptor mRNAs were detected in the patient 's tumor and the series of 17 additional pheochromocytomas . This study shows that pheochromocytomas express functional 5-HT(4) receptors that are responsible for the stimulatory action of metoclopramide on catecholamine- and granin-derived peptide secretion . All 5-HT(4) receptor agonists must therefore be contraindicated in patients with proven or suspected pheochromocytoma . P25021 overexpression induces U937 cell differentiation despite triggered mechanisms to attenuate DB02527 signalling . Knowing that cell-surface receptors that recognize and respond to extracellular stimuli are key components for the regular communication between individual cells required for the survival of any living organism , the aim of the present work was to investigate the effect of P25021 overexpression on the U937 signal transduction pathway and its consequences on cell proliferation and differentiation . The overexpression of P25021 led to an increase in DB02527 basal levels , a leftward shift of agonist concentration-response curves , and similar maximal response to agonist treatment , suggesting that overexpressed H2Rs act as functional spare receptors . In this system cells triggered several mechanisms tending to restore DB02527 basal levels to those of the naïve cells . P25021 overexpression induced PDE activity stimulation and P25098 overexpression . In spite of the onset of these regulatory mechanisms , H2 agonist and rolipram treatments induced the terminal differentiation of the P25021 overexpressed clone , conversely to the naïve cells . Present findings show that stably P25021 overexpression alters DB02527 signalling as the result of not only the amounts of second messenger generated but also the activation or upregulation of various components of signalling cascade , leading to an adapted biologically unique system . This adaptation may represent an advantage or a disadvantage , depending on the biological system , but in any case , the existence of compensatory mechanisms should be considered when a clinical treatment is designed . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Gene expression profiles of adipose tissue of obese rats after central administration of neuropeptide Y- Q15761 antisense oligodeoxynucleotides by cDNA microarrays . To investigate the gene expression profiles of adipose tissue of obese rats after central administration of neuropeptide Y- Q15761 antisense oligodeoxynucleotides ( ODNs ) , Q15761 antisense , mismatched ODNs or vehicle was intracerebroventricularly injected and cDNA microarrays were undertaken . Central administration of Q15761 antisense ODNs decreased food intake , body weight and serum insulin compared with both vehicle and mismatched ODNs . The average area of adipocytes both at retroperitoneal and epididymal adipose tissue were fall in antisense group while only the weight of the retroperitoneal fat pats was reduced in antisense group . cDNA microarrays containing 18,000 genes/Ests were used to investigate gene expression of adipose tissue . Autoradiographic analysis showed that 404 , 81 , and 34 genes were differently expressed over twofold , threefold , and fivefold , respectively . The analysis of gene expression profiles indicated that 332 genes were up-regulated and 187 genes were down-regulated in response to Q15761 antisense ODNs treatment . Different clusters of genes associated with apoptosis , signal transduction , energy metabolism , lipid metabolism , etc. , such as P51114 , Q8WV24 , Q7L5Y9 , P27986 , P13598 , Q00169 , P62158 , Q13557 , P61925 , P14416 , O95258 , CKB , P22760 , P38571 , O15254 , O60427 , were concerned . Analysis of differentially expressed genes will help to understand the effects of Q15761 antisense ODNs therapy . The human interleukin 18 gene Q14116 maps to 11q22.2-q22.3 , closely linked to the P14416 gene locus and distinct from mapped IDDM loci . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Effects of enhancement and antagonism of 5-hydroxytryptamine activity on the influence of metoclopramide on gastric emptying . This study examines the influence of the serotonergic system on the effect of metoclopramide on gastric emptying . Six subjects received the following pretreatments before metoclopramide and paracetamol : fluoxetine ( 5-HT uptake inhibitor ) ; meterogoline ( 5-HT1 antagonist ) ; pizotifen ( 5-HT2 antagonist ) or methysergide ( 5-HT1 and 5-HT2 antagonist ) . One regimen consisted of metoclopramide ( 5- Q9H205 antagonist and Q13639 agonist ) alone . Gastric emptying was measured by the mean cumulative fraction absorbed-time profiles of paracetamol . Methysergide/metoclopramide significantly delayed gastric emptying from 30 min onwards . DB01233 with either metergoline or pizotifen did not retard gastric emptying to the same extent , suggesting a greater influence with simultaneous 5-HT1 and 5HT2 blockade . DB01233 /fluoxetine caused a significant decrease in the fractional absorption of paracetamol at 5 min when compared to the metoclopramide regimen . It was assumed that the influence of metoclopramide was not optimal at this stage , therefore possibly indicating domination of 5- Q9H205 over Q13639 effects , resulting in gastric delay . It therefore seems as if all the 5-HT receptors present in the gut have a role to play in the control of gastric emptying .

MH_train_17

MH_train_17

MH_train_17

interacts_with DB09030?

multiple_choice

Thrombin induces slug-mediated P12830 transcriptional repression and the parallel up-regulation of P19022 by a transcription-independent mechanism in Q96AT9 cells . The proliferation , directional migration to the vitreous and epithelial-mesenchymal transition ( EMT ) of quiescent , differentiated retinal pigment epithelium ( Q96AT9 ) cells is a major feature in the development of proliferative vitreoretinopathy ( P15151 ) following exposure of the immuno-privileged eye niche to serum components , thrombin among them . We have previously documented thrombin induction of Q96AT9 cell proliferation and migration . We here analyzed the effect of thrombin on the E/N cadherin switch , a hallmark of EMT . Results show that thrombin induces the specific repression of epithelial P12830 gene transcription , alongside with the up-regulation of mesenchymal P19022 protein in Q96AT9 cells . We demonstrate , for the first time , that thrombin induces P12830 repression by stimulating snail-2 ( O43623 ) transcription factor expression , and the concomitant up-regulation of P19022 through the transcription-independent increase in protein translation promoted by PI3K/PKC-ζ/ P42345 signaling . Our present findings suggest that the activation of protease-activated receptor-1 ( P25116 ) by thrombin induces EMT of Q96AT9 cells , further supporting a central role for thrombin in P15151 pathogenesis . Protease-activated receptors upregulate cyclooxygenase-2 expression in human endothelial cells . We have previously shown that the serine protease thrombin and other G protein-coupled agonists acutely enhance synthesis and release of prostacyclin from human umbilical vein endothelial cells ( HUVEC ) through activation of P47712 alpha . Here , we show that thrombin and other physiological endothelial cell agonists upregulate P35354 induction in HUVEC . Thrombin treatment caused a rapid and sustained increase in prostacyclin ( DB01240 ) synthesis from HUVEC . Thrombin and a selective protease-activated receptor-1 ( P25116 ) peptide ( TRAP ) evoked dose- and time-dependent increases in P35354 protein expression which were equivalent to that induced by the proinflammatory cytokine P01583 . Quantitative and real-time PCR analysis showed enhanced P35354 mRNA expression in thrombin- or TRAP-stimulated HUVEC whereas P23219 expression was unaffected . A P55085 agonist peptide also induced P35354 protein and mRNA expression with kinetics distinct from those of thrombin , and promoted DB01240 release . These results demonstrate that regulation of P35354 induction is an important functional response of HUVEC to PAR activation and suggest that PARs promote sustained upregulation of prostanoid production in human endothelium . Clinical potential of vorapaxar in cardiovascular risk reduction in patients with atherosclerosis . DB09030 ( ZONTIVITY™ , formerly known as P35240 530348 ) is a specific , orally active antagonist of the protease-activated receptor-1 ( P25116 ) on platelets . It inhibits thrombin-induced platelet activation by binding to the ectodomain of P25116 . After animal studies and Phase II studies showed that vorapaxar sufficiently inhibits platelet activation without significantly increasing bleeding complications , safety and efficacy of vorapaxar were assessed in two large multicenter trials in patients with coronary artery disease and atherosclerosis . The Thrombin-Receptor Antagonist for Clinical Event Reduction in Acute Coronary Syndromes ( TRACER ) trial investigated safety and efficacy of vorapaxar in patients with an acute coronary syndrome without ST-segment elevation . The Trial to Assess the Effects of DB09030 in Preventing Heart Attack and Stroke in Patients With Atherosclerosis-Thrombolysis In Myocardial Infarction 50 ( TRA 2°P-TIMI 50 ) investigated atherothrombotic events in patients with stable atherosclerosis . Results of both studies suggested that vorapaxar given in addition to standard antiplatelet therapy can reduce atherothrombotic events , but increases the risk of mild and moderate bleeding complications . This review article summarizes the main results of TRACER and TRA 2°P-TIMI 50 and suggests patient cohorts that might benefit from treatment with vorapaxar in addition to standard antiplatelet therapy . DB09030 : a novel protease-activated receptor-1 inhibitor . INTRODUCTION : Platelet activation and reactivity are pivotal for both acute and chronic atherothrombotic event occurrences . AREAS COVERED : Only 20 % relative risk ( ∼ 2 % absolute risk ) reduction associated with newer P2Y(12) receptor blocker therapy such as prasugrel and ticagrelor compared with clopidogrel indicates that dual antiplatelet therapy may be associated with a ceiling effect in attenuating platelet-mediated ischemic event occurrence and that residual ischemic event occurrences are mediated by other pathways that are unblocked by current antiplatelet therapy . Therefore , inhibition of the thrombin-protease-activated receptor ( PAR ) -1 interaction may provide additional benefits in attenuating ischemic event occurrence in selected patients . There are two major P25116 blockers are under investigations - vorapaxar and atopaxar . In preclinical and Phase I - II studies , inhibition of thrombin-mediated platelet activation by a P25116 inhibitor , in general , has added to the antithrombotic efficacy of aspirin and clopidogrel without increasing bleeding . However , intracranial hemorrhage in patients with a history of stroke associated with vorapaxar and hepatic toxicity associated with atopaxar are important concerns . EXPERT OPINION : At this time , the specific role of P25116 inhibitor in the settings of percutaneous coronary intervention and acute coronary syndrome , both during the acute setting and as a long-term therapeutic agent , is not clear . Although the P25116 inhibitors are associated with less bleeding , its effectiveness as an antithrombotic agent and also side effects are major concerns . Future large-scale trials with goals addressing these concerns are needed to define the specific role of P25116 receptor inhibitor . P01308 -like growth factor-1 potentiates platelet activation via the P41252 / P42336 pathway . As insulin-like growth factor-1 ( DB01277 ) is present in the alpha granules of platelets and its receptor is expressed on the platelet surface , it may contribute to the amplification of platelet responses and pathogenesis of cardiovascular disease . The functional and signaling pathways that are involved in DB01277 modulation of platelet function , however , are presently unknown . Here , I report that DB01277 stimulation of platelets results in dose-dependent phosphorylation of the IGF receptor in the range of 1 to 100 nM . Phosphorylation of the IGF receptor is rapid and sustained , with maximal phosphorylation reached within 1 minute . Furthermore , DB01277 stimulates tyrosine phosphorylation of insulin receptor substrate-1 ( P35568 ) and Q9Y4H2 and their association with the p85 subunit of phosphoinositide-3 kinase ( PI3K ) . DB01277 -stimulated tyrosine phosphorylation of P35568 and Q9Y4H2 and subsequent p85 binding is transient and precedes phosphorylation of protein kinase B ( P31749 ) on Ser473 . P25116 -mediated platelet aggregation is potentiated by DB01277 and this potentiation , together with P31749 phosphorylation , is abolished by the P42336 inhibitors PI-103 and PIK-75 . Importantly , the IGF receptor inhibitor DB00238 -AEW541 and the neutralization antibody alphaIR3 inhibit SFLLRN-stimulated aggregation , implicating DB01277 in autocrine regulation of platelet function . These results demonstrate that DB01277 activates the IGF receptor/ P41252 /PI3K/ P31749 pathway , and that P42336 is essential for the potentiatory effect of DB01277 on platelet responses . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . No differences in the pharmacodynamics and pharmacokinetics of the thrombin receptor antagonist vorapaxar between healthy Japanese and Caucasian subjects . BACKGROUND : DB09030 , a novel antiplatelet agent in advanced clinical development for the prevention and treatment of atherothrombotic disease , is a potent , orally bioavailable thrombin receptor antagonist selective for the protease-activated receptor 1 ( P25116 ) . METHODS : Since race/ethnicity may affect the safety , efficacy and dosage of drugs , this study was conducted to evaluate potential differences in the pharmacodynamics , pharmacokinetics and safety of vorapaxar after single ( 5 , 10 , 20 , or 40 mg ) or multiple ( 0.5 , 1 , or 2.5 mg once daily ) doses in healthy Japanese and matched ( gender , age , height , and weight ) Caucasian volunteers . RESULTS : DB09030 was well tolerated in both Japanese and Caucasian subjects . Pharmacodynamic and pharmacokinetic profiles of vorapaxar in the two racial/ethnic groups were similar . In both racial groups , complete inhibition of platelet aggregation was achieved most rapidly with vorapaxar 40 mg and was consistently achieved and maintained with a 2.5 mg daily maintenance dose . CONCLUSION : There were no substantial differences in the safety , pharmacokinetics or pharmacodynamics of vorapaxar between Japanese and Caucasian subjects . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Porphyromonas gingivalis selectively up-regulates the HIV-1 coreceptor P51681 in oral keratinocytes . Primary infection of oral epithelial cells by HIV-1 , if it occurs , could promote systemic infection . Most primary systemic infections are associated with R5-type HIV-1 targeting the R5-specific coreceptor P51681 , which is not usually expressed on oral keratinocytes . Because coinfection with other microbes has been suggested to modulate cellular infection by HIV-1 , we hypothesized that oral keratinocytes may up-regulate P51681 in response to the oral endogenous pathogen Porphyromonas gingivalis by cysteine-protease ( gingipains ) activation of the protease-activated receptors ( PARs ) or LPS signaling through the TLRs . The OKF6/ O14746 -2-immortalized normal human oral keratinocyte line expressed P61073 , whereas P51681 was not detectable . When exposed to P. gingivalis ATCC 33277 , O14746 -2 cells induced greater time-dependent expression of P51681 -specific mRNA and surface coreceptors than P61073 . By comparing arg- ( Rgp ) and lys-gingipain ( Kgp ) mutants , a mutant deficient in both proteases , and the action of trypsin , P. gingivalis Rgp was strongly suggested to cleave P25116 and P55085 to up-regulate P51681 . P51681 was also slightly up-regulated by an isogenic gingipain-deficient mutant , suggesting the presence of a nongingipain-mediated mechanism . Purified P. gingivalis LPS also up-regulated P51681 . Blocking O60603 and O00206 receptors with Abs attenuated induction of P51681 , suggesting LPS signaling through TLRs . P. gingivalis , therefore , selectively up-regulated P51681 by two independent signaling pathways , Rgp acting on P25116 and P55085 , and LPS on O60603 and O00206 . By inducing P51681 expression , P. gingivalis coinfection could promote selective R5-type HIV-1 infection of oral keratinocytes . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . DB09030 , an oral P25116 receptor antagonist , does not affect the pharmacokinetics and pharmacodynamics of warfarin . PURPOSE : DB09030 is an orally active protease-activated receptor 1 ( P25116 ) antagonist that inhibits thrombin-induced platelet aggregation . This open-label study assessed the pharmacokinetics and pharmacodynamics of single-dose warfarin in the presence/absence of multiple-dose vorapaxar in 12 healthy men . METHODS : Subjects received two treatments separated by ≥ 7-day washout : Treatment A warfarin 25 mg ( Day 1 ) ; Treatment B vorapaxar 2.5 mg/day on Days 1-6 and vorapaxar 40 mg coadministered with warfarin 25 mg ( Day 7 ) . R-warfarin , S-warfarin , and prothrombin time ( PT ) were assayed predose and up to 120 h postdose . RESULTS : The geometric mean ratio ( GMR ) as a percentage ( warfarin + vorapaxar/warfarin ) was calculated . The GMR ( 90 % CIs ) estimates of C(max) were 105 ( 99 , 111 ) and 105 ( 99 , 112 ) for R- and S-warfarin , respectively . The GMR ( 90 % CIs ) estimates of AUC(0-∞) were 108 ( 101 , 116 ) and 105 ( 96 , 115 ) for R- and S-warfarin , respectively . The GMR ( 95 % CIs ) estimates of AUC ( 0-120 h ) for PT and INR were 97 ( 95 , 98 ) and 96 ( 94 , 98 ) , respectively . CONCLUSION : Results of this study indicate that vorapaxar has no meaningful effect on the pharmacokinetics or pharmacodynamics of warfarin , suggesting that the coadministration of these two drugs or vorapaxar coadministered with other P11712 / P33261 substrates is unlikely to cause a clinically significant pharmacokinetic drug interaction . P37840 activates microglia by inducing the expressions of matrix metalloproteinases and the subsequent activation of protease-activated receptor-1 . The mutation or overexpression of alpha-synuclein protein plays a pivotal role in the pathogenesis of Parkinson 's disease . In our preliminary experiments , we found that alpha-synuclein induced the expression of matrix metalloproteinases ( MMPs ) ( P03956 , -3 , -8 , and -9 ) in rat primary cultured microglia . Thus , the current study was undertaken to determine the roles of MMPs in alpha-synuclein-induced microglial activation . The inhibition of P08254 , -8 , or -9 significantly reduced NO and reactive oxygen species levels and suppressed the expression of P01375 and IL-1beta . Notably , P22894 inhibitor suppressed P01375 production more efficaciously than P08254 or P14780 inhibitors . Inhibition of P08254 or -9 also suppressed the activities of MAPK , NF-kappaB , and AP-1 . Previously , protease-activated receptor-1 ( P25116 ) has been associated with the actions of MMPs , and thus , we further investigated the role of P25116 in alpha-synuclein-induced inflammatory reactions . A P25116 -specific inhibitor and a P25116 antagonist significantly suppressed cytokine levels , and NO and reactive oxygen species production in alpha-synuclein-treated microglia . Subsequent P25116 cleavage assay revealed that P08254 , -8 , and -9 , but not alpha-synuclein , cleaved the synthetic peptide containing conventional P25116 cleavage sites . These results suggest that MMPs secreted by alpha-synuclein-stimulated microglia activate P25116 and amplify microglial inflammatory signals in an autocrine or paracrine manner . Furthermore , our findings suggest that modulation of the activities of MMPs and/or P25116 may provide a new therapeutic strategy for Parkinson 's disease . Pharmacodynamics and pharmacokinetics of the novel P25116 antagonist vorapaxar ( formerly P35240 530348 ) in healthy subjects . PURPOSE : The aim of our study was to evaluate the pharmacology of vorapaxar ( P35240 530348 ) , an oral P25116 antagonist , in healthy volunteers . METHODS AND RESULTS : In two randomized , placebo-controlled studies , subjects received either single ascending doses of vorapaxar ( 0.25 , 1 , 5 , 10 , 20 , or 40 mg ; n = 50 ) , multiple ascending doses of vorapaxar ( 1 , 3 , or 5 mg/day for 28 days ; n = 36 ) , a loading dose ( 10 or 20 mg ) followed by daily maintenance doses ( 1 mg ) for 6 days ( n = 12 ) , or placebo . Single 20- and 40-mg doses of vorapaxar completely inhibited thrombin receptor activating peptide ( TRAP ) -induced platelet aggregation ( > 80 % inhibition ) at 1 h and sustained this level of inhibition for ≥72 h . Multiple doses yielded complete inhibition on Day 1 ( 5 mg/day ) and Day 7 ( 1 and 3 mg/day ) . Adverse events were generally mild , transient , and unrelated to dose . CONCLUSION : DB09030 provided rapid and sustained dose-related inhibition of platelet aggregation without affecting bleeding or clotting times . DB09030 : first global approval . DB09030 [ DB09030 (®) ( US ) ] , an orally active protease-activated receptor-1 ( P25116 ) receptor antagonist , has been developed by Merck & Co for the reduction of thrombotic cardiovascular events in patients with a history of myocardial infarction ( MI ) or peripheral arterial disease ( PAD ) . DB09030 has received its first global approval for this indication in the US . This article summarizes the milestones in the development of vorapaxar leading to this first approval for the reduction of thrombotic cardiovascular events in patients with a prior MI or PAD . P00747 -induced aggregation of PANC-1 cells requires conversion to plasmin and is inhibited by endogenous plasminogen activator inhibitor-1 . PANC-1 cells express proteinase-activated receptors ( PARs ) -1 , -2 , and respond to their activation by transient elevation of cytosolic [ Ca(2+) ] and accelerated aggregation ( Wei et al. , 2006 , J Cell Physiol 206:322-328 ) . We studied the effect of plasminogen ( Q9UQ90 ) , an inactive precursor of the P25116 -activating protease , plasmin ( PN ) on aggregation of pancreatic adenocarcinoma ( PDAC ) cells . A single dose of Q9UQ90 time- and dose-dependently promoted PANC-1 cells aggregation in serum-free medium , while PN did not . PANC-1 cells express urokinase plasminogen activator ( uPA ) , which continuously converted Q9UQ90 to PN . This activity and Q9UQ90 -induced aggregation were inhibited by the uPA inhibitor amiloride . Q9UQ90 -induced aggregation was also inhibited by alpha-antiplasmin and by the PN inhibitor epsilon-aminocaproic acid ( DB00513 ) . Direct assay of uPA activity revealed very low rate , markedly enhanced in the presence of Q9UQ90 . Moreover , in Q9UQ90 activator inhibitor 1-deficient PANC-1 cells , uPA activity and Q9UQ90 -induced aggregation were markedly potentiated . Two additional human PDAC cell lines , MiaPaCa and Colo347 , were assayed for Q9UQ90 -induced aggregation . Both cell lines responded by aggregation and exhibited Q9UQ90 -enhanced uPA activity . We hypothesized that the continuous conversion of Q9UQ90 to PN by endogenous uPA is limited by PN 's degradation and negatively controlled by endogenously produced P05121 . Indeed , we found that PANC-1 cells inactivate PN with t1/2 of approximately 7 h , while the continuous addition of PN promoted aggregation . Our data suggest that PANC-1 cells possess intrinsic , P05121 -sensitive mechanism for promotion of aggregation and differentiation by prolonged exposure to Q9UQ90 and , possibly , additional precursors of PARs agonists . Safety of the novel protease-activated receptor-1 antagonist vorapaxar in Japanese patients with a history of ischemic stroke . BACKGROUND : DB09030 , formerly P35240 530348 , is a novel , orally active , potent thrombin receptor inhibitor selective for the protease-activated receptor-1 ( P25116 ) . Previous phase II studies of patients undergoing urgent or scheduled percutaneous coronary intervention treated with vorapaxar plus aspirin and clopidogrel or ticlopidine showed a trend toward reducing major adverse cardiac events , particularly myocardial infarction , without increasing bleeding risk . The present study evaluated the safety of vorapaxar in Japanese patients with a history of ischemic stroke receiving aspirin . METHODS : Ninety patients with previous ischemic stroke ( ≥14 days to < 1 year before randomization ) were randomized to receive vorapaxar ( 1 or 2.5 mg ) or placebo once daily for 60 days . All patients received aspirin ( 75-150 mg/day ) . The primary endpoint was overall incidence of adverse events during the protocol-defined treatment phase ( 60 days ) . RESULTS : Addition of vorapaxar to aspirin did not significantly increase the overall incidence of adverse events , including serious adverse events . None of the patients treated with vorapaxar plus aspirin experienced thrombolysis in myocardial infarction major or minor bleeding versus 1 patient treated with placebo . Nonfatal stroke occurred in 1 patient allocated to placebo and 1 patient allocated to vorapaxar . CONCLUSIONS : DB09030 used in combination with standard doses of aspirin was safe and well tolerated in Japanese subjects with a history of ischemic stroke . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Molecular determinants of trastuzumab efficacy : What is their clinical relevance ? DB00072 -containing therapy is a standard of care for human epidermal growth factor receptor-2 ( P04626 ) -positive breast cancer . In pre-clinical models , a wide range of molecular mechanisms have been associated with reduced sensitivity to trastuzumab in vitro . These include expression of the truncated P04626 receptor fragment p95HER2 , activating mutation of the gene encoding the class 1A catalytic subunit of phosphatidylinositol 3-kinase ( P42336 ) , loss of phosphatase and tensin homolog ( P60484 ) , activation of other downstream signal transducers , prevention of cell cycle arrest , increased signaling through alternative ( HER or non-HER ) tyrosine kinase receptors , and resistance to antibody-dependent cellular cytotoxicity . However , the clinical significance of these mechanisms as determinants of trastuzumab efficacy in vivo has been unclear . Here , we review clinical studies of potential predictive biomarkers of trastuzumab efficacy in P04626 -positive breast cancer and consider whether evaluation of such markers might inform patient selection for therapy . We find that clinical evidence relating to potential predictive biomarkers is mostly limited to small , retrospective studies , many of which have yielded conflicting findings . Some trends are evident in the retrospective data and in biomarker analyses from randomized clinical trials , particularly relating to activation of the phosphatidylinositol 3-kinase pathway , but none is sufficiently strong to form a basis for patient selection . This may be explained by the fact that multiple mechanisms of action determine the clinical efficacy of trastuzumab . In the absence of novel , validated biomarkers of efficacy , trastuzumab eligibility should continue to be based on evaluation of P04626 status according to standard methods . Comparison of gene expression profiles and related pathways in chronic thromboembolic pulmonary hypertension . Chronic thromboembolic pulmonary hypertension ( CTEPH ) is one of the main causes of severe pulmonary hypertension . However , despite treatment ( pulmonary endarterectomy ) , in approximately 15-20 % of patients , pulmonary vascular resistance and pulmonary arterial pressure continue to increase . To date , little is known about the changes that occur in gene expression in CTEPH . The identification of genes associated with CTEPH may provide insight into the pathogenesis of CTEPH and may aid in diagnosis and treatment . In this study , we analyzed the gene expresion profiles of pulmonary artery endothelial cells from 5 patients with CTEPH and 5 healthy controls using oligonucleotide microarrays . Bioinformatics analyses using the Gene Ontology ( GO ) and KEGG databases were carried out to identify the genes and pathways specifically associated with CTEPH . Signal transduction networks were established to identify the core genes regulating the progression of CTEPH . A number of genes were found to be differentially expressed in the pulmonary artery endothelial cells from patients with CTEPH . In total , 412 GO terms and 113 pathways were found to be associated with our list of genes . All differential gene interactions in the Signal-Net network were analyzed . P52333 , P30679 , O15264 , P32121 and P25116 were the most significantly altered . Bioinformatics analysis may help gather and analyze large amounts of data in microarrays by means of rigorous experimental planning , scientific statistical analysis and the collection of complete data . In this study , a novel differential gene expression pattern was constructed . However , further studies are required to identify novel targets for the diagnosis and treatment of CTEPH . Pharmacokinetics of the novel P25116 antagonist vorapaxar in patients with hepatic impairment . PURPOSE : To determine whether hepatic impairment has an effect on the pharmacokinetics ( PK ) of vorapaxar or M20 , its main pharmacologically active metabolite . METHODS : This was an open-label study in which a single 40-mg oral dose of vorapaxar was administered to patients with mild ( n = 6 ) , moderate ( n = 6 ) , and severe ( n = 4 ) hepatic impairment and healthy controls ( n = 16 ) matched for age , gender , weight , and height . Blood samples for vorapaxar and M20 assay were collected predose and at frequent intervals up to 8 weeks postdose . RESULTS : Plasma vorapaxar and M20 PK profiles were similar between patients with impaired liver function and healthy controls . Group mean values for vorapaxar C(max) and AUC(tf) were 206-279 ng/mL and 14,200-18,200 ng·h/mL , respectively , with the lowest values observed in patients with severe impairment . DB09030 median T(max) and mean t(1/2) values were 1.00-1.75 h and 298-366 h , respectively . There was no apparent correlation between vorapaxar or M20 exposure or t(1/2) values and disease severity . DB09030 was generally well tolerated ; one serious adverse event ( gastrointestinal bleeding secondary to ruptured esophageal varices ) was reported in a patient with severe hepatic impairment . CONCLUSIONS : Hepatic impairment had no clinically relevant effect on the PK of vorapaxar and M20 . No dose or dosage adjustment of vorapaxar will be required in patients with mild to moderate hepatic impairment . Although systemic exposure to vorapaxar does not appear to increase in patients with severe hepatic impairment , administration of vorapaxar to such patients is not recommended given their bleeding diathesis . Effect of curcumin on nuclear factor kappaB signaling pathways in human chronic myelogenous K562 leukemia cells . Curcumin , a natural product isolated from the plant Curcuma longa , has a diverse range of molecular targets that influence numerous biochemical and molecular cascades . Curcumin has been shown to inhibit nuclear factor kappaB ( NF-kappaB ) activation at several steps in the NF-kappaB signaling pathways and thereby controls numerous NF-kappaB-regulated genes involved in various diseases . In the present study , we investigated the effect of curcumin pretreatment on 84 tumor necrosis factor-alpha ( P01375 ) -activated genes of NF-kappaB pathways in K562 cells , using a real-time PCR array . Our results show that transcription of 29 NF-kappaB-related mRNAs was significantly downregulated ( Q9Y239 , P13500 , P25942 , P04141 , P25116 , P05362 , O14920 , Q14164 , P01583 , P01584 , P05231 , P10145 , O43187 , Q9UDY8 , Q13233 , Q99836 , P19838 , Q00653 , P25963 , P35813 , P04049 , Q01201 , P42224 , O15455 , P01375 , TNFalphaIP3 , P50591 , and Q8IUC6 ) , whereas 10 mRNAs were induced ( AGT , P29466 , P09919 , P01100 , P01579 , P22301 , Q86XR7 , O60603 , Q9NR96 , and P26842 ) . Western blot analysis of P25942 , P19838 ( p50 ) , Q01201 , P25963 ( P25963 ) , and P22301 as well as an P10145 secretion assay confirmed our results . Taken together , we show that curcumin regulates an impressive number of NF-kappaB genes within the different NF-kappaB signaling pathways . Genetic association between P43250 /β-arrestin 2 and dopamine supersensitivity psychosis in schizophrenia . BACKGROUND/AIM : Dopamine supersensitivity psychosis ( P15924 ) , clinically characterized by unstable and severe psychosis or tardive dyskinesia and often categorized as treatment-resistant schizophrenia , is promoted by long-term antipsychotic treatment . An upregulation of the dopamine D2 receptor caused by antipsychotic(s) is involved in the development of P15924 . The present study explored the potential roles of P43250 ( P43250 ) and β-arrestin 2 ( P32121 ) that are involved in the trafficking of P14416 in patients with P15924 . METHODS : We conducted a genetic association study of P43250 / P32121 between the patients with P15924 episodes [ P15924 (+) group : N=108 ] and the patients without P15924 (-) episodes [ P15924 (-) group : N=169 ] from the total group of patients ( N=333 ) . Based on the patients ' treatment history , a P15924 episode was defined as withdrawal psychosis , developed tolerance to antipsychotic effect , and tardive dyskinesia ( the remaining 56 patients were excluded due to insufficient information ) . RESULTS : The results revealed that none of the allelic or genotyping distributions of five single nucleotide polymorphisms ( SNPs ) of P43250 and three SNPs of P32121 showed any significant difference between the P15924 (+) and P15924 (-) groups . CONCLUSION : The results suggest that the SNP analyses of these two molecules fail to classify patients into the potential clinical subtype of P15924 (+) or P15924 (-) group . However , since P43250 and P32121 are surely involved in dopamine D2 receptor metabolism , further studies based on prospective observations of the onset of P15924 under specific antipsychotic treatments are needed . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . P25116 antagonists : current state of evidence . DB09030 ( P35240 530348 ) and atopaxar ( E5555 ) are oral protease-activated receptor-1 ( P25116 ) antagonists with high bioavailability . They inhibits thrombin induced platelet aggregation by competitively inhibiting P25116 . We systematically evaluated the evidence for the efficacy and safety of all P25116 antagonists as well as for the individual drugs vorapaxar and atopaxar in different databases-PubMed , EMBASE , Scopus , and Cochrane register of Controlled Clinical Trials (CENTRAL).We selected randomized controlled trials of P25116 antagonists that reported on cardiovascular mortality as a clinical outcome . The random-effects Mantel-Haenszel model was used to evaluate the effect of P25116 antagonists on cardiovascular mortality . Seven trials were selected ( N = 42,355 ) for analysis . P25116 antagonists as a class , as well as individually , were associated with a non-significant numerically lower risk of cardiovascular mortality than that seen with agents used in the control group ; RR , 0.93 ; 95 % CI , 0.83-1.04 ; P = 0.20 ) . No heterogeneity was noted . However , P25116 antagonists also appeared to increase the risk of bleeding significantly . P25116 antagonists appear to be associated with some reduction in the risk of cardiovascular mortality ; however the significantly higher bleeding risk noted with P25116 antagonists appear to mandate a very careful selection of patients that may benefit without a substantially increased risk of bleeds . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . DB09030 expands antiplatelet options . Which patients may benefit from thrombin receptor antagonism ? DB09030 is the first substance of a new class of antiplatelet drugs that has been tested in large clinical trials . The protease-activated receptor 1 ( P25116 ) antagonist inhibits thrombin-induced platelet activation to prevent atherothrombosis . In the phase 3 trials TRACER ( acute coronary syndrome ) and TRA 2P-TIMI 50 ( stable atherosclerosis ) reducing ischemic events with vorapaxar came at the cost of bleeding . TRACER compared vorapaxar to placebo in 12,944 patients who had non-ST-segment elevation acute coronary syndromes on top of contemporary treatment including dual antiplatelet therapy ( aspirin and clopidogrel ) . DB09030 reduced ischemic events non-significantly , but increased bleeding significantly , therefore not justifying triple antiplatelet therapy in this setting . Follow-up was stopped early because of bleeding . TRA 2P-TIMI 50 examined 26,449 patients who had a history of myocardial infarction , ischemic stroke , or peripheral arterial disease . DB09030 reduced ischemic events and increased bleeding both significantly . Recruitment of patients with prior stroke was stopped early . Net clinical outcome and subgroup analyses suggested that vorapaxar could be beneficial for patients with prior myocardial infarction - but no history of stroke . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . 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 .

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A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Single-walled carbon nanotubes ( SWCNTs ) enhance DB00761 - , acetylcholine- , and serotonin-induced contractions and evoke oxidative stress on rabbit ileum . We examined the effects of intravenous administration of purified arc-discharge single-walled carbon nanotubes ( SWCNTs ) on rabbit ileum to establish the possibility of using these SWCNTs as cell markers or drug carriers for the treatment of intestinal diseases . The SWCNT purification process eliminated carbonaceous impurities and decreased the amount of metals . SWCNTs increased the contractile responses induced by DB00761 , acetylcholine ( ACh ) , and serotonin ( 5-HT ) in rabbit ileum . Verapamil , apamin , glibenclamide , quinine and charybdotoxin reduced the contractile responses induced by ACh and 5-HT in ileum from rabbits treated with SWCNTs , indicating that voltage-dependent Ca2+ channels and small , intermediate , and large-conductance Ca(2+)-activated , DB00171 -sensitive , and voltage-dependent K+ channels are involved in these effects . Atropine and hexamethonium reduced the ACh response , indicating that muscarinic and nicotinic receptors are involved in this effect . DB00904 and GR 113808 reduced the 5-HT response , indicating that serotonin 5- Q9H205 and Q13639 receptors are involved in this effect . SWCNTs increased the malondialdehyde plus 4-hydroxyalkenals and carbonyl levels in rabbit plasma and ileum , indicating that SWCNTs produce oxidative stress . SWCNTs did not produce relevant histological changes or modify the levels of the inflammatory mediators P35228 and P35354 in the ileum . In conclusion , this study demonstrates that the intravenous administration of SWCNTs can evoke oxidative stress and affect contractility in rabbit ileum . These effects could reduce the possibility of using the arc-discharge SWCNTs as cell markers or drug carriers to treat intestinal diseases . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . O76074 inhibitors enhance celecoxib killing in multiple tumor types . The present studies determined whether clinically relevant phosphodiesterase 5 ( O76074 ) inhibitors interacted with a clinically relevant NSAID , celecoxib , to kill tumor cells . Celecoxib and O76074 inhibitors interacted in a greater than additive fashion to kill multiple tumor cell types . Celecoxib and sildenafil killed ex vivo primary human glioma cells as well as their associated activated microglia . Knock down of O76074 recapitulated the effects of O76074 inhibitor treatment ; the nitric oxide synthase inhibitor L-NAME suppressed drug combination toxicity . The effects of celecoxib were P35354 independent . Over-expression of O15519 -s or knock down of CD95/ Q13158 significantly reduced killing by the drug combination . CD95 activation was dependent on nitric oxide and ceramide signaling . CD95 signaling activated the JNK pathway and inhibition of JNK suppressed cell killing . The drug combination inactivated P42345 and increased the levels of autophagy and knock down of Beclin1 or Q9H1Y0 strongly suppressed killing by the drug combination . The drug combination caused an ER stress response ; knock down of IRE1α/ P17861 enhanced killing whereas knock down of eIF2α/ P18848 / P35638 suppressed killing . DB00203 and celecoxib treatment suppressed the growth of mammary tumors in vivo . Collectively our data demonstrate that clinically achievable concentrations of celecoxib and sildenafil have the potential to be a new therapeutic approach for cancer . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Basal cell carcinoma and variants in genes coding for immune response , DNA repair , folate and iron metabolism . Basal cell carcinoma ( BCC ) is one of the most common neoplasms in the world and its incidence has been increasing worldwide in recent years . BCCs are caused by an interplay between genetic and environment factors . We conducted a case-control association study in BCC patients and controls from Sweden and Finland . Fifteen single nucleotide polymorphisms ( SNPs ) , P05231 -174G/C , -634G/C , and -597G/A ; P22301 -1082G/A and -592C/A ; IL-1beta-511C/T ; NBS1 exon 5 Glu185Gln ; Q01831 exon 15 Lys939Gln ; P18074 exon 23 Lys751Gln ; P18887 exon 10 Arg399Gln ; O43542 exon 7 Thr241Met ; cyclin D1 exon 4 G870A ; P42898 exon 4 Ala222Val and exon 7 Glu429Ala ; Q30201 exon 4 C282Y were performed by Pyrosequencing and RFLP techniques . Most of the genotype distributions were in accordance with the Hardy-Weinberg equilibrium ( HWE ) , except for P22301 -1082G/A , where cases with BCC showed a significant deviation from HWE ( P = 0.04 ) . Linkage disequilibrium was observed between the -174 and -597 alleles in the P05231 gene in the present populations . No difference between BCC and controls appeared in any of the SNPs analyzed . Only the combined distributions of TT/AA genotypes in P42898 exon 4 ( C/T ) and exon 7 ( A/C ) showed slight increase in BCC compared to controls ( P < 0.07 , OR : 1.94 ; 95 % CI : 0.96-3.89 ) . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Synthesis and biological evaluation of novel pyrrolidine-2,5-dione derivatives as potential antidepressant agents . Part 1 . A series of 3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives was synthesized and their biological activity was evaluated . The chemical structures of the newly prepared compounds were confirmed by (1)H NMR , (13)C NMR and P19957 -HRMS spectra data . All tested compounds proved to be potent P08908 receptor and serotonin transporter protein ( P31645 ) ligands . Among them , compounds 15 , 18 , 19 and 30 showed significant affinity for P08908 and P31645 . Computer docking simulations carried out for compounds 15 , 31 and 32 to models of P08908 receptor and P31645 confirm the results of biological tests . Due to high affinity for the P08908 receptor and moderate affinity for P31645 , compounds 31 , 32 , 35 , and 37 were evaluated for their affinity for D2L , P50406 , P34969 and 5- Q13049 receptors . In vivo tests , in turn , resulted in determining the functional activity of compounds 15 , 18 , 19 and 30 to the P08908 receptor . The results of these tests indicate that all of the ligands possess properties characteristic of P08908 receptor agonists . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Evaluation of the endogenous cannabinoid system in mediating the behavioral effects of dipyrone ( metamizol ) in mice . DB04817 is a common nonopioid analgesic and antipyretic , which , in many countries , is available over the counter and is more widely used than paracetamol or aspirin . However , the exact mechanisms by which dipyrone acts remain inconclusive . Two novel arachidonoyl-conjugated metabolites are formed in mice following the administration of dipyrone that are dependent on the activity of fatty acid amide hydrolase ( FAAH ) , which also represents the major catabolic enzyme of the endogenous cannabinoid ligand anandamide . These arachidonoyl metabolites not only inhibit cyclooxygenase ( P23219 / P35354 ) but also bind to cannabinoid receptors at low micromolar concentrations . The relative contributions of cannabinoid receptors and FAAH in the overall behavioral response to dipyrone remain untested . Accordingly , the two primary objectives of the present study were to determine whether the behavioral effects of dipyrone would ( a ) be blocked by cannabinoid receptor antagonists and ( b ) occur in FAAH mice . Here , we report that thermal antinociceptive , hypothermic , and locomotor suppressive actions of dipyrone are mediated by a noncannabinoid receptor mechanism of action and occurred after acute or repeated administration irrespective of FAAH . These findings indicate that FAAH-dependent arachidonoyl metabolites and cannabinoid receptors are not requisites by which dipyrone exerts these pharmacological effects under noninflammatory conditions . Differential effects of endotoxin and fibrinogen degradation products ( P14324 ) on liver synthesis of fibrinogen and albumin : evidence for the involvement of a novel monokine in the stimulation of fibrinogen synthesis induced by P14324 . 1. Administration of endotoxin or fibrinogen degradation products ( FDPs ) in rats increase fibrinogen synthesis comparable to that found during the acute phase response . 2 . An increased fibrinogen synthesis is also found in co-cultures of hepatocytes with peripheral blood mononuclear cells upon administration of endotoxin or FDPs , but not in primary cultures of hepatocytes alone . 3 . However , the increased synthesis of fibrinogen by FDPs is not accompanied by a decreased albumin synthesis , as in the case of stimulated fibrinogen synthesis induced by endotoxin in vivo and in co-cultures of hepatocytes with peripheral blood mononuclear cells , or induced by monocytic products in vivo and in primary cultures of hepatocytes alone . 4 . Since IL-1 and/or P05231 could not be accounted for the stimulation of fibrinogen synthesis without a decreased albumin synthesis , a novel monokine produced by mononuclear cells upon Q9NRC9 administration might be involved . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Novel bioactive metabolites of dipyrone ( metamizol ) . DB04817 is a common antipyretic drug and the most popular non-opioid analgesic in many countries . In spite of its long and widespread use , molecular details of its fate in the body are not fully known . We administered dipyrone orally to mice . Two unknown metabolites were found , viz. the arachidonoyl amides of the known major dipyrone metabolites , 4-methylaminoantipyrine ( 2 ) and 4-aminoantipyrine ( 3 ) . They were identified by P19957 -LC-MS/MS after extraction from the CNS , and comparison with reference substances prepared synthetically . The arachidonoyl amides were positively tested for cannabis receptor binding ( CB(1) and CB(2) ) and cyclooxygenase inhibition ( P23219 and P35354 in tissues and as isolated enzymes ) , suggesting that the endogenous cannabinoid system may play a role in the effects of dipyrone against pain . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Screening of 134 single nucleotide polymorphisms ( SNPs ) previously associated with type 2 diabetes replicates association with 12 SNPs in nine genes . More than 120 published reports have described associations between single nucleotide polymorphisms ( SNPs ) and type 2 diabetes . However , multiple studies of the same variant have often been discordant . From a literature search , we identified previously reported type 2 diabetes-associated SNPs . We initially genotyped 134 SNPs on 786 index case subjects from type 2 diabetes families and 617 control subjects with normal glucose tolerance from Finland and excluded from analysis 20 SNPs in strong linkage disequilibrium ( r(2) > 0.8 ) with another typed SNP . Of the 114 SNPs examined , we followed up the 20 most significant SNPs ( P < 0.10 ) on an additional 384 case subjects and 366 control subjects from a population-based study in Finland . In the combined data , we replicated association ( P < 0.05 ) for 12 SNPs : P37231 Pro12Ala and His447 , Q14654 Glu23Lys and rs5210 , P01375 -857 , P11168 Ile110Thr , P20823 /TCF1 rs2701175 and GE117881_360 , P35558 -232 , Q13562 Thr45Ala , P05231 -598 , and P22413 Lys121Gln . The replication of 12 SNPs of 114 tested was significantly greater than expected by chance under the null hypothesis of no association ( P = 0.012 ) . We observed that SNPs from genes that had three or more previous reports of association were significantly more likely to be replicated in our sample ( P = 0.03 ) , although we also replicated 4 of 58 SNPs from genes that had only one previous report of association . Design of novel potent antihyperlipidemic agents with antioxidant/anti-inflammatory properties : exploiting phenothiazine 's strong antioxidant activity . Because atherosclerosis is an inflammatory process involving a series of pathological events such as dyslipidemia , oxidative stress , and blood clotting mechanisms , we hereby report the synthesis and evaluation of novel compounds in which antioxidant , anti-inflammatory , and squalene synthase ( P37268 ) inhibitory/hypolipidemic activities are combined in simple molecules through design . The coupling of two different pharmacophores afforded compounds 1-12 , whose biological profile was markedly improved compared to those of parent lead structures ( i.e. , the hypolipidemic 2-hydroxy-2-aryl-(benzo)oxa ( or thia ) zine and the antioxidant phenothiazine ) . Most derivatives strongly inhibited in vitro microsomal lipid and LDL peroxidation , exhibiting potent free-radical scavenging activity . They further significantly inhibited P37268 activity and showed remarkable antidyslipidemic activity in vivo in animal models of acute and high-fat-induced hyperlipidemia . Finally , several compounds showed anti-inflammatory activity in vitro , inhibiting cycloxygenase ( P23219 /2 ) activity . The multimodal properties of the new compounds and especially their combined antioxidant/ P37268 / P36551 inhibitory activity render them interesting lead compounds for further evaluation against atherosclerosis . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Altered gene expression by low-dose arsenic exposure in humans and cultured cardiomyocytes : assessment by real-time PCR arrays . Chronic arsenic exposure results in higher risk of skin , lung , and bladder cancer , as well as cardiovascular disease and diabetes . The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array ( TLDA ) . We found that expression of tumor necrosis factor-α ( P01375 -α ) , which activates both inflammation and NF-κB-dependent survival pathways , was strongly associated with water and urinary arsenic levels . Expression of P22460 , which encodes a potassium ion channel protein , was positively associated with water and toe nail arsenic levels . Expression of 2 and 11 genes were positively associated with nail and urinary arsenic , respectively . Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans , we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro . Expression of the ion-channel genes CACNA1 , Q12809 , P51787 and P15382 were down-regulated by 1-μM arsenic . Alteration of some common pathways , including those involved in oxidative stress , inflammatory signaling , and ion-channel function , may underlay the seemingly disparate array of arsenic-associated diseases , such as cancer , cardiovascular disease , and diabetes . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Activation of the P19957 -K/Akt pathway mediates cGMP enhanced-neurogenesis in the adult progenitor cells derived from the subventricular zone . The intracellular mechanisms that regulate neurogenesis remain unclear . Using neurospheres isolated from the subventricular zone ( SVZ ) of the adult rat , we investigated the effect of cyclic guanosine monophosphate ( cGMP ) and its signaling pathway on the induction of neurogenesis . Neurospheres expressed phosphodiesterase 5 ( O76074 ) and treatment of neurospheres with DB00203 , a specific inhibitor of O76074 , significantly increased cGMP levels and neurogenesis . In addition , incubation of neurospheres with DB00203 significantly phosphorylated Akt , which was associated with an increase of phosphorylation of glycogen synthase kinase 3 ( GSK-3 ) , a downstream target of Akt . Coincubation of neurospheres with DB00203 and LY 294002 , a pharmacological inhibitor of P19957 -K/Akt , abolished DB00203 -induced phosphorylated Akt and GSK-3 . Furthermore , LY 294002 blocked DB00203 -increased SVZ cell proliferation . These data suggest that DB00203 -enhanced neurogenesis likely occurs through activation of the P19957 -K/Akt/GSK-3 pathway . A comparative study of the antipyretic effects of indomethacin and dipyrone in rats . OBJECTIVE : Compare the antipyretic effects of dipyrone and indomethacin . MATERIALS AND METHODS : Fever was induced in rats by i. v. LPS or i . c . v. interleukins ( IL ) , prostaglandins ( PG ) , arachidonic acid ( AA ) , pre-formed pyrogenic factor ( PFPF ) , tumour necrosis factor-alpha ( P01375 ) or corticotrophin releasing hormone ( P06850 ) . DB04817 and indomethacin were administered i.p. , arginine vasopressin V1-receptor antagonist , d(CH2)5 DB00135 (Me)AVP , into the ventral septal area . Cyclooxygenase ( P23219 /-2 ) blocking activity was assessed in transfected COS-7 cells . P06850 release from isolated hypothalami was determined by ELISA . RESULTS : Indomethacin or dipyrone reduced LPS , IL-1beta , P05231 or P01375 induced fever and P06850 release from rat hypothalamus . Only dipyrone inhibited P10145 , PFPF or PGF2alpha fever . Only indomethacin inhibited fever induced by AA or IL-1beta , plus AA . Neither antipyretic affected fever caused by DB00917 or P06850 . d(CH2)5Tyr(Me)AVP only blocked antipyresis induced by indomethacin . DB04817 at a very high concentration ( 10 mM ) inhibited only P23219 , while indomethacin ( 0.1 microM ) blocked P23219 and P35354 in COS-7 cells . CONCLUSION : The antipyretic effect of dipyrone differs from that of indomethacin in that it does not depend on AVP release or inhibition of PG synthesis . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Celecoxib blocks cardiac Kv1.5 , Kv4.3 and Kv7.1 ( P51787 ) channels : effects on cardiac action potentials . Celecoxib is a P35354 inhibitor that has been related to an increased cardiovascular risk and that exerts several actions on different targets . The aim of this study was to analyze the effects of this drug on human cardiac voltage-gated potassium channels ( Kv ) involved on cardiac repolarization Kv1.5 ( I(Kur) ) , Kv4.3+ Q9NS61 ( I(to1) ) and Kv7.1+ P15382 ( I(Ks) ) and to compare with another P35354 inhibitor , rofecoxib . Currents were recorded in transfected mammalian cells by whole-cell patch-clamp . Celecoxib blocked all the Kv channels analyzed and rofecoxib was always less potent , except on Kv4.3+ Q9NS61 channels . Kv1.5 block increased in the voltage range of channel activation , decreasing at potentials positive to 0 mV . The drug modified the activation curve of the channels that became biphasic . Block was frequency-dependent , increasing at fastest frequencies . Celecoxib effects were not altered by DB08837 (out) in R487Y mutant Kv1.5 channels but the kinetics of block were slower and the degree of block was smaller with DB08837 (in) , indicating that celecoxib acts from the cytosolic side . We confirmed the blocking properties of celecoxib on native Kv currents from rat vascular cells , where Kv1.5 are the main contributors ( IC(50)≈ 7 μM ) . Finally , we demonstrate that celecoxib prolongs the action potential duration in mouse cardiac myocytes and shortens it in guinea pig cardiac myocytes , suggesting that Kv block induced by celecoxib may be of clinical relevance . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . P35354 regulates the proliferation of glioma stem like cells . Cancer stem-like cells ( CSCs ) possessing features of neural precursor cells ( NPC ) influence initiation , recurrence and chemoresistance of glioblastoma multiforme ( GBM ) . As inflammation is crucial for glioblastoma progression we investigated the effect of chronic IL-1β treatment on CSCs derived from glioblastoma cell line U87MG . Exposure to IL-1β for 10 days increased ( i ) accumulation of 8-OHdG - a key biomarker of oxidative DNA damage ; ( ii ) DNA damage response ( DDR ) indicators γ P16104 , Q13315 and DNA-PK ; ( iii ) nuclear and cytoplasmic p53 and P35354 levels and ( iv ) interaction between P35354 and p53 . Despite upregulating p53 expression IL-1β had no effect on cell cycle progression , apoptosis or self renewal capacity of CSCs . P35354 inhibitor Celecoxib reduced self renewal capacity and increased apoptosis of both control and IL-1β treated CSCs . Therefore the ability of P35354 to regulate proliferation of CSCs irrespective of exposure to IL-1β , warrants further investigation of P35354 as a potential anti-glioma target .

MH_train_19

MH_train_19

MH_train_19

interacts_with DB00083?

multiple_choice

Accelerated intoxication of GABAergic synapses by botulinum neurotoxin A disinhibits stem cell-derived neuron networks prior to network silencing . Botulinum neurotoxins ( BoNTs ) are extremely potent toxins that specifically cleave SNARE proteins in peripheral synapses , preventing neurotransmitter release . Neuronal responses to BoNT intoxication are traditionally studied by quantifying SNARE protein cleavage in vitro or monitoring physiological paralysis in vivo . Consequently , the dynamic effects of intoxication on synaptic behaviors are not well-understood . We have reported that mouse embryonic stem cell-derived neurons ( ESNs ) are highly sensitive to BoNT based on molecular readouts of intoxication . Here we study the time-dependent changes in synapse- and network-level behaviors following addition of DB00083 to spontaneously active networks of glutamatergic and GABAergic ESNs . Whole-cell patch-clamp recordings indicated that DB00083 rapidly blocked synaptic neurotransmission , confirming that ESNs replicate the functional pathophysiology responsible for clinical botulism . Quantitation of spontaneous neurotransmission in pharmacologically isolated synapses revealed accelerated silencing of GABAergic synapses compared to glutamatergic synapses , which was consistent with the selective accumulation of cleaved P60880 at Q99259 (+) pre-synaptic terminals at early timepoints . Different latencies of intoxication resulted in complex network responses to DB00083 addition , involving rapid disinhibition of stochastic firing followed by network silencing . Synaptic activity was found to be highly sensitive to P60880 cleavage , reflecting the functional consequences of the localized cleavage of the small subpopulation of P60880 that is engaged in neurotransmitter release in the nerve terminal . Collectively these findings illustrate that use of synaptic function assays in networked neurons cultures offers a novel and highly sensitive approach for mechanistic studies of toxin:neuron interactions and synaptic responses to BoNT . A fluorescent ligand-binding alternative using Tag-lite® technology . G-protein-coupled receptors ( GPCRs ) are crucial cell surface receptors that transmit signals from a wide range of extracellular ligands . Indeed , 40 % to 50 % of all marketed drugs are thought to modulate GPCR activity , making them the major class of targets in the drug discovery process . Binding assays are widely used to identify high-affinity , selective , and potent GPCR drugs . In this field , the use of radiolabeled ligands has remained so far the gold-standard method . Here the authors report a less hazardous alternative for high-throughput screening ( HTS ) applications by the setup of a nonradioactive fluorescence-based technology named Tag-lite(®) . Selective binding of various fluorescent ligands , either peptidic or not , covering a large panel of GPCRs from different classes is illustrated , particularly for chemokine ( P61073 ) , opioid ( δ , µ , and κ ) , and cholecystokinin ( CCK1 and CCK2 ) receptors . Affinity constants of well-known pharmacological agents of numerous GPCRs are in line with values published in the literature . The authors clearly demonstrate that the Tag-lite binding assay format can be successfully and reproducibly applied by using different cellular materials such as transient or stable recombinant cells lines expressing P60880 -tagged GPCR . Such fluorescent-based binding assays can be performed with adherent cells or cells in suspension , in 96- or 384-well plates . Altogether , this new technology offers great advantages in terms of flexibility , rapidity , and user-friendliness ; allows easy miniaturization ; and makes it completely suitable for HTS applications . P60880 is essential for cortical granule exocytosis in mouse eggs . Synaptosome-associated protein of 25 kDa ( P60880 ) has been shown to play an important role in Ca2+-dependent exocytosis in neurons and endocrine cells . During fertilization , sperm-egg fusion induces cytosolic Ca2+ mobilization and subsequently Ca2+-dependent cortical granule ( CG ) exocytosis in eggs . However , it is not yet clear whether P60880 is involved in this process . In this study , we determined the expression and function of P60880 in mouse eggs . mRNA and P60880 were detected in metaphase II ( MII ) mouse eggs by RT-PCR and immunoblot analysis , respectively . Next , to determine the function of P60880 , we evaluated the change in CG exocytosis with a membrane dye , tetramethylammonium-1,6-diphenyl-1,3,5-hexatriene , after microinjection of a botulinum neurotoxin A ( DB00083 ) , which selectively cleaves P60880 in MII eggs . Sperm-induced CG exocytosis was significantly inhibited in the DB00083 -treated eggs . The inhibition was attenuated by coinjection of P60880 . These results suggest that P60880 may be involved in Ca2+-dependent CG exocytosis during fertilization in mouse eggs . Unique substrate recognition by botulinum neurotoxins serotypes A and E . Botulinum neurotoxins ( BoNTs ) are zinc proteases that cleave SNARE proteins to elicit flaccid paralysis by inhibiting the fusion of neurotransmitter-carrying vesicles to the plasma membrane of peripheral neurons . There are seven serotypes of BoNT , termed A-G . BoNT serotype A and serotype E cleave P60880 at residues 197-198 and 180-181 , respectively . Unlike other zinc proteases , the BoNTs recognize extended regions of P60880 for cleavage . The basis for this extended substrate recognition and specificity is unclear . Saturation mutagenesis and deletion mapping identified residues 156-202 of P60880 as the optimal cleavage domain for DB00083 , whereas the optimal cleavage domain for BoNT/E was shorter , comprising residues 167-186 of P60880 . Two sub-sites were resolved within each optimal cleavage domain , which included a recognition or active site ( AS ) domain that contained the site of cleavage and a binding ( B ) domain , which contributed to substrate affinity . Within the AS domains , the P1 ' , P09131 , and Q15084 sites of P60880 contributed to scissile bond cleavage by LC/A , whereas the P1 ' and P2 sites of P60880 contributed to scissile bond cleavage by LC/E . These studies provide insight into the development of strategies for small molecule inhibitors of the BoNTs . [ COMPARATIVE STUDY OF NIGROSTRIATAL SYSTEMS IN WISTAR RATS AND RATS PRONE TO SEIZURES ] . In this work we analyzed the levels of functional activity of dopaminergic , GABA-ergic and glutamatergic neurons in the nigrostriatal system of control Wistar rats and Krushinsky-Molodkina ( KM ) rats prone to audiogenic seizures . In KM rats we have revealed disturbed activity of GABA- and dopaminergic neurons in substania nigra whereas the level of glutamatergic neurotransmission remained unchanged . We have also observed no significant differences in Q05329 /67 and phospho-tyrosine hydroxylase contents in the striatum of KM and control Wistar rats . However , a high level of D1 dopamine receptor and a decreased level of D2 receptor found can mediate the upregulation of glutamatergic neurotransmission . Indeed , the expression of vesicular glutamate transporter type 2 ( VGlut2 ) and Q13224 subunit of DB01221 receptor was increased in the striatum of KM rats . In striatal glutamatergic fibers phosphorylated P27361 /2 kinases have been revealed ; at the same time , in KM rats an increased P27361 /2 activity has been detected both in striatum and substantia nigra . This finding correlated with activation of exocytosis rate as evidenced by downregulation of P60880 level . Apart from other reasons , the activation of glutamatergic system may be a result of disruption of the inhibitory effect of the dopamine- and GABAergic systems of substantia nigra that innervate striatum . We suppose that the increased activity of striatal glutamatergic neurons of KM rats without an adequate inhibition by GABA- and dopaminergic systems may be one of the reasons of high convulsive susceptibility in KM rats . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal . Botulinum neurotoxin A impairs neurotransmission following retrograde transynaptic transport . The widely used botulinum neurotoxin A ( DB00083 ) blocks neurotransmission via cleavage of the synaptic protein P60880 ( synaptosomal-associated protein of 25 kDa ) . Recent evidence demonstrating long-distance propagation of P60880 proteolysis has challenged the idea that DB00083 remains localized to the injection site . However , the extent to which distant neuronal networks are impacted by DB00083 retrograde trafficking remains unknown . Importantly , no studies have addressed whether P60880 cleavage translates into structural and functional changes in distant intoxicated synapses . Here we show that the DB00083 injections into the adult rat optic tectum result in P60880 cleavage in retinal neurons two synapses away from the injection site , such as rod bipolar cells and photoreceptors . Retinal endings displaying cleaved P60880 were enlarged and contained an abnormally high number of synaptic vesicles , indicating impaired exocytosis . Tectal injection of DB00083 in rat pups resulted in appearance of truncated- P60880 in cholinergic amacrine cells . Functional imaging with calcium indicators showed a clear reduction in cholinergic-driven wave activity , demonstrating impairments in neurotransmission . These data provide the first evidence for functional effects of the retrograde trafficking of DB00083 , and open the possibility of using DB00083 fragments as drug delivery vehicles targeting the central nervous system . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Second generation steroidal 4-aminoquinolines are potent , dual-target inhibitors of the botulinum neurotoxin serotype A metalloprotease and P. falciparum malaria . Significantly more potent second generation 4-amino-7-chloroquinoline ( 4,7-ACQ ) based inhibitors of the botulinum neurotoxin serotype A ( DB00083 ) light chain were synthesized . Introducing an amino group at the C(3) position of the cholate component markedly increased potency ( IC50 values for such derivatives ranged from 0.81 to 2.27 μM ) . Two additional subclasses were prepared : bis(steroidal)-4,7-ACQ derivatives and bis(4,7-ACQ)cholate derivatives ; both classes provided inhibitors with nanomolar-range potencies ( e.g. , the Ki of compound 67 is 0.10 μM ) . During DB00083 challenge using primary neurons , select derivatives protected P60880 by up to 89 % . Docking simulations were performed to rationalize the compounds ' in vitro potencies . In addition to specific residue contacts , coordination of the enzyme 's catalytic zinc and expulsion of the enzyme 's catalytic water were a consistent theme . With respect to antimalarial activity , the compounds provided better IC90 activities against chloroquine resistant ( CQR ) malaria than CQ , and seven compounds were more active than mefloquine against CQR strain W2 . Enhancement of the endopeptidase activity of botulinum neurotoxin by its associated proteins and dithiothreitol . Botulinum neurotoxins type A ( DB00083 ) , the most toxic substance known to man , is produced by Clostridium botulinum type A as a complex with a group of neurotoxin-associated proteins ( NAPs ) , possibly through a polycistronic expression of a clustered group of genes . The botulinum neurotoxin complex is the only known example of a protein complex where a group of proteins ( NAPs ) protect another protein ( BoNT ) against acidity and proteases of the GI tract . We now report that NAPs also potentiate the DB01593 endopeptidase activity of DB00083 in both in vitro and in vivo assays against its known intracellular target protein , 25 kDa synaptosomal associated protein ( P60880 ) . While DB00083 exhibited no protease activity prior to reduction with dithiothreitol ( DTT ) , the DB00083 complex exhibited a high protease activity even in its nonreduced form . Our results suggest that the bacterial production of NAPs along with BoNT is designed for the NAPs to play an accessory role in the neurotoxin function , in contrast to their previously known limited role in protecting the neurotoxin in the GI tract and in the external environment . Structural features of DB00083 change considerably upon disulfide reduction , as revealed by near-UV circular dichroism spectroscopy . DB00083 in the reduced form adopts a more flexible structure than in the unreduced form , as also indicated by large differences in DeltaH values ( 155 vs 248 kJ mol-1 ) of temperature-induced unfolding of DB00083 . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . Long-lasting effects of elevated neonatal leptin on rat hippocampal function , synaptic proteins and DB01221 receptor subunits . The high circulating levels of leptin in neonatal rodents do not seem to be regulating energy balance at this age , but rather may play an important role for brain development . We tested the hypothesis that high neonatal leptin levels modify hippocampal function and production of synaptic proteins with possible long-term consequences on long-term potentiation ( LTP ) in adulthood . We first showed that in postnatal day ( P01160 ) 10 neonates , acute leptin treatment functionally activated leptin receptors ( ObR ) in the P00915 and DG regions of the hippocampus through the induction of phosphoERK1/2 , but not phosphoSTAT3 protein although both phospho-proteins were induced in the arcuate nucleus . We next examined whether chronic leptin administration ( 3 mg/kg BW , intraperitoneally ) during the first 2 weeks of life ( postnatal day , P01160 2-14 ) produces a functional signal in the hippocampus that alters the expression of DB01221 receptor subunits ( Q9UHB4 , Q12879 , Q13224 ) , synaptic proteins and LTP in the short and long-term . In P01160 10 as in adults ( P01160 70 ) rats , chronic leptin treatment increased Q9UHB4 expression in the hippocampus while reducing Q13224 protein levels . Elevated hippocampal concentrations of synapsin2A and synaptophysin were detected during leptin treatment on P01160 10 suggesting increased neurotransmitter release . In adults , only P60880 expression was increased after neonatal leptin treatment . LTP was reduced dramatically by leptin treatment in preweaning rats although the changes did not persist until adulthood . Elevated exposure to leptin during a critical period of neonatal hippocampal development might serve to enhance DB01221 -dependent functions other than LTP and have important effects on synaptogenesis and neurotransmitter release . The destructive effect of botulinum neurotoxins on the SNARE protein : P60880 and synaptic membrane fusion . Synaptic exocytosis requires the assembly of syntaxin 1A and P60880 on the plasma membrane and synaptobrevin 2 ( P63027 ) on the vesicular membrane to bridge the two opposite membranes . It is believed that the three SNARE proteins assemble in steps along the dynamic assembly pathway . The C-terminus of P60880 is known to be the target of botulinum neurotoxins ( DB00083 and BoNT/E ) that block neurotransmitters release in vivo . In this study , we employed electron paramagnetic resonance ( EPR ) spectroscopy to investigate the conformation of the P60880 C-terminus in binary and ternary SNARE complexes . The fluorescence lipid mixing assay shows that the C-terminal of P60880 is essential for membrane fusion , and that the truncated P60880 mutants cleaved by DB00083 and BoNT/E display different inhibition effects on membrane fusion : P60880 -25E ( Δ26 ) abolishes the fusion activity of the SNARE complex , while P60880 -25A ( Δ9 ) loses most of its function , although it can still form a SDS-resistant SNARE complex as the wild-type P60880 . CW-EPR spectra validate the unstable structures of the SNARE complex formed by P60880 mutants . We propose that the truncated P60880 mutants will disrupt the assembly of the SNARE core complex , and then inhibit the synaptic membrane fusion accordingly . Botulinum neurotoxin A and neurotoxin E cleavage products of synaptosome-associated protein of 25 kd exhibit distinct actions on pancreatic islet beta-cell Kv2.1 channel gating . OBJECTIVES : Synaptosome-associated protein of 25 kd ( P60880 ) regulates pancreatic islet beta-cell-delayed rectifier K channels ( Kv2.1 ) in addition to insulin exocytosis . Botulinum neurotoxin A ( DB00083 ) and E ( BoNT/E ) cleavage and presumed deletion of P60880 have been used to examine P60880 function . We hypothesized that proteolytic products of P60880 ( 206 amino acids ) resulting from DB00083 and BoNT/E cleavage , P60880 (1-197) and P60880 (1-180) , have independent actions on beta-cell Kv gating . METHODS : We examined by confocal microscopy and immunoblotting DB00083 and BoNT/E cleavage of P60880 to these N-terminal fragments , and the consequent effects of these BoNTs and P60880 fragments on Kv currents in rat beta cells and MIN6 cells by patch clamp electrophysiology . RESULTS : Confocal microscopy and immunoblotting showed that MIN6 cells transfected with DB00083 or BoNT/E generated P60880 (1-197) and P60880 (1-180) fragments that were retained in the cytosol . Both BoNTs caused increased rate of channel activation and slowed channel inactivation , mimicked by these P60880 fragments , but not full-length P60880 . These P60880 fragments potentiated tetraethylammonium block of beta-cell Kv currents . CONCLUSIONS : DB00083 or BoNT/E treatment of beta cells generates N-terminal P60880 fragments that are retained in beta cells to directly influence Kv channel gating in a manner distinct from full-length P60880 , contributing to overall actions of these BoNTs on insulin secretion . Inhibitory effect of botulinum toxin type A on the NANC system in rat respiratory models of neurogenic inflammation . This study investigated whether botulinum toxin type A ( DB00083 ) inhibits respiratory neurogenic inflammation in the non-adrenergic , non-cholinergic ( NANC ) transmitter system in rats . Neurogenic inflammation models were induced in Sprague Dawley ( SD ) rats through bilateral cerebral artery occlusion ( BCAO ) for different times ( 0 , 30 and 60 min ) or by stimulation with capsaicin at different doses ( 5 or 15 g/kg ) . Pre-Bötzinger Complex-Spikes and the expression of DB05875 , synaptosomal-associated protein-25 ( P60880 ) , and reactive oxygen species ( ROS ) were detected with or without pretreatment of rats with DB00083 ( 15 or 30 U/kg ) . BCAO reduced pre-Bot C spike activity ( spike/s ) and increased the breath rate ( breaths/s ) in an unstable pattern in comparison to controls , while pretreatment with DB00083 slightly reduced this phenomenon . Pretreatment with DB00083 inhibited BCAO- or capsaicin-induced increases in expression of P60880 , DB05875 , and ROS in a dose-dependent manner in brainstem and lung tissue . DB00083 exerts a suppressive effect on neurogenic inflammation via non-adrenergic , non-cholinergic transmitters . These results add to the body of evidence elucidating the non-cholinergic effects of DB00083 in the context of neurogenic inflammation . Botulinum neurotoxin A activity is dependent upon the presence of specific gangliosides in neuroblastoma cells expressing synaptotagmin I . Botulinum neurotoxin A ( DB00083 ) is the deadliest of all known biological substances . Although its toxicity makes DB00083 a biological warfare threat , its biologic activity makes it an increasingly useful therapeutic agent for the treatment of muscular disorders . However , almost 200 years after its discovery , the neuronal cell components required for the activity of this deadly toxin have not been unequivocally identified . In this work , neuroblastoma cells expressing synaptotagmin I , a protein shown to be bound by DB00083 , were used to determine whether specific gangliosides were necessary for DB00083 activity as measured by synaptosomal-associated protein of 25 kDa ( P60880 ) cleavage . Ganglioside GT1b was found to support DB00083 activity significantly more effectively than GD1a , which was far more effective than GM1 when added to ganglioside-deficient murine cholinergic Neuro 2a or to human adrenergic SK-N-SH neuroblastoma cells . Whereas both cell lines expressed synaptotagmin I , P60880 cleavage was not observed in the absence of complex gangliosides . These results indicate that 1 ) gangliosides are required for DB00083 activity , 2 ) synaptotagmin I in the absence of gangliosides does not support DB00083 activity , and 3 ) Neuro 2a cells are an efficient model system for studying the biological activity of DB00083 . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . DB00435 -cGMP-protein kinase G signaling pathway induces anoxic preconditioning through activation of DB00171 -sensitive K+ channels in rat hearts . DB00435 ( NO ) plays an important role in anoxic preconditioning to protect the heart against ischemia-reperfusion injuries . The present work was performed to study better the NO-cGMP-protein kinase G ( PKG ) signaling pathway in the activation of both sarcolemmal and mitochondrial DB00171 -sensitive K+ ( KATP ) channels during anoxic preconditioning ( P25054 ) and final influence on reducing anoxia-reperfusion ( A/R ) -induced cardiac damage in rat hearts . The upstream regulating elements controlling NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection were investigated . The involvement of both inducible and endothelial NO synthases ( P35228 and P29474 ) in the progression of this signaling pathway was followed . Final cellular outcomes of ischemia-induced injury after different preconditioning in the form of lactate dehydrogenase release , DNA strand breaks , and malondialdehyde formation as indexes of cell injury and lipid peroxidation , respectively , were investigated . The lactate dehydrogenase and malondialdehyde values decreased in the groups that underwent preconditioning periods with specific mitochondrial KATP channels opener diazoxide ( 100 microM ) , nonspecific mitochondrial KATP channels opener pinacidil ( 50 microM ) , S-nitroso-N-acetylpenicillamine ( P60880 , 300 microM ) , or beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclicmonophosphorothioate , Sp-isomer ( 10 microM ) before the A/R period . Preconditioning with P60880 significantly reduced the DNA damage . The effect was blocked by glibenclamide ( 50 microM ) , 5-hydroxydecanoate ( 100 microM ) , NG-nitro-L-arginine methyl ester ( 200 microM ) , and beta-phenyl-1,N2-etheno-8-bromoguanosine-3',5'-cyclic monophosphorothioate , Rp-isomer ( 1 microM ) . The results suggest P35228 , rather than P29474 , as the major contributing NO synthase during P25054 treatment . Moreover , the PKG shows priority over NO as the upstream regulator of NO-cGMP-PKG signal-induced KATP channel opening that leads to cardioprotection during P25054 treatment . Mechanism of substrate recognition by botulinum neurotoxin serotype A . Botulinum neurotoxins ( BoNTs ) are zinc proteases that cleave SNARE proteins to elicit flaccid paralysis by inhibiting neurotransmitter-carrying vesicle fusion to the plasma membrane of peripheral neurons . Unlike other zinc proteases , BoNTs recognize extended regions of P60880 for cleavage ; however , the molecular basis for this extended substrate recognition is unclear . Here , we define a multistep mechanism for recognition and cleavage of P60880 by DB00083 . P60880 initially binds along the belt region of DB00083 , which aligns the Q15084 residue to the S5 pocket at the periphery of the active site . Although the exact order of each step of recognition of P60880 by DB00083 at the active site is not clear , the initial binding could subsequently orient the P4'-residue of P60880 to form a salt bridge with the S4'-residue , which opens the active site allowing the P1'-residue access to the S1'-pocket . Subsequent hydrophobic interactions between the P09131 residue of P60880 and the S3 pocket optimize alignment of the scissile bond for cleavage . This explains how the BoNTs recognize and cleave specific coiled SNARE substrates and provides insight into the development of inhibitors to prevent botulism . Novel chimeras of botulinum and tetanus neurotoxins yield insights into their distinct sites of neuroparalysis . Botulinum neurotoxin ( BoNT ) A or E and tetanus toxin ( TeTx ) bind to motor-nerve endings and undergo distinct trafficking ; their light-chain ( LC ) proteases cleave soluble N-ethylmaleimide-sensitive factor attachment protein receptors ( SNAREs ) peripherally or centrally and cause flaccid or spastic paralysis , respectively . To seek protein domains responsible for local blockade of transmitter release ( BoNTs ) rather than retroaxonal transport to spinal neurons ( TeTx ) , their acceptor-binding moieties ( H(C) ) -- or in one case , heavy chain ( HC ) -- were exchanged by gene recombination . Each chimera , expressed and purified from Escherichia coli , entered rat cerebellar neurons to cleave their substrates , blocked in vitro nerve-induced muscle contractions , and produced only flaccid paralysis in mice . Thus , the local cytosolic delivery of DB00083 or BoNT/E proteases and the contrasting retrograde transport of TeTx are not specified solely by their HC or H(C) ; DB00083 LC translocated locally irrespective of being targeted by either of the latter TeTx domains . In contrast , BoNT/E protease fused to a TeTx enzymatically inactive mutant ( TeTIM ) caused spastic paralysis and cleaved P60880 in spinal cord but not the injected muscle . Apparently , TeTIM precludes cytosolic release of BoNT/E protease at motor nerve endings . It is deduced that the LCs of the toxins , acting in conjunction with HC domains , dictate their local or distant destinations . P00734 haplotype associated with kidney stone disease in Northeastern Thai patients . OBJECTIVE : To evaluate genetic variations associated with kidney stone disease in Northeastern Thai patients . METHODS : Altogether , 67 single nucleotide polymorphisms ( SNP ) distributed within 8 candidate genes , namely P04155 , P05109 , P06702 , P80511 , P02760 , P10451 , P07911 , and F2 , which encode stone inhibitor proteins , including trefoil factor 1 , calgranulin ( A , B , and C ) , bikunin , osteopontin , tamm-Horsfall protein , and prothrombin , respectively , were initially genotyped in 112 individuals each and in additional subjects to consist of 164 patients and 216 control subjects in total . RESULTS : We found that minor allele and homozygous genotype frequencies of 8 of 10 SNPs distributed within the F2 gene were significantly higher in the control group than in the patient group . Two F2 haplotypes were found to be dually associated with kidney stone risk , one ( TGCCGCCGCG ) with increased disease risk and the other ( CGTTCCGCTA ) with decreased disease risk . However , these 2 haplotypes were associated with the disease risks in only the female , not the male , group . CONCLUSIONS : The results of our study indicate that genetic variation of F2 is associated with kidney stone risk in Northeastern Thai female patients . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . Neuromuscular transmission and muscle contractility in P60880 -deficient coloboma mice . Synaptosomal associated protein of 25 kDa ( P60880 ) is a cytoplasmic protein that participates in the docking and fusion of synaptic vesicles with the nerve terminal in preparation for neurotransmitter release . P60880 is also a substrate for three of the seven serotypes of botulinum neurotoxin ( BoNT ) . Intoxication by DB00083 , /C1 or /E results in weakness and paralysis of skeletal muscle due to cleavage of P60880 ( and syntaxin la in the case /C1 ) at discrete serotype-specific sites . To elucidate the role of P60880 in muscle function in more detail , contractility and neuromuscular transmission were studied in a mutant mouse model termed coloboma . The coloboma mutation results from a contiguous deletion of 1-2 centiMorgans on chromosome 2 , which includes the entire P60880 locus and three other identified genes . Homozygotes do not survive beyond gestation day 6 ; heterozygotes ( Cm/+ ) have a normal life-span but express reduced levels of P60880 mRNA and protein in the brain . The consequences of the Cm/+ mutation on twitch and tetanic tension , quantal release of neurotransmitter and spinal motoneuron expression of P60880 were examined in the present study . Contrary to expectations , Cm/+ mice exhibited no alteration in twitch tension and generated normal tetanic tension even at the highest frequency examined ( 800 Hz ) . Microelectrode recordings revealed that MEPP amplitude and frequency were both within control limits . The ventral spinal cord of Cm/+ mice showed no deficiency in P60880 content and immunohistochemical examination of nerve terminals in Cm/+ mice disclosed that P60880 levels and distribution were similar to those of control mice . It is concluded that spinal motor neurons up-regulate P60880 to preserve vital neuromuscular function . Proteolysis of P60880 isoforms by botulinum neurotoxin types A , C , and E : domains and amino acid residues controlling the formation of enzyme-substrate complexes and cleavage . Tetanus toxin and the seven serologically distinct botulinal neurotoxins ( DB00083 to BoNT/G ) abrogate synaptic transmission at nerve endings through the action of their light chains ( L chains ) , which proteolytically cleave VAMP ( vesicle-associated membrane protein ) /synaptobrevin , P60880 ( synaptosome-associated protein of 25 kDa ) , or syntaxin . BoNT/C was reported to proteolyze both syntaxin and P60880 . Here , we demonstrate that cleavage of P60880 occurs between Arg198 and Ala199 , depends on the presence of regions Asn93 to Glu145 and Ile156 to Met202 , and requires about 1,000-fold higher L chain concentrations in comparison with DB00083 and BoNT/E . Analyses of the DB00083 and BoNT/E cleavage sites revealed that changes in the carboxyl-terminal residues , in contrast with changes in the amino-terminal residues , drastically impair proteolysis . A proteolytically inactive DB00083 L chain mutant failed to bind to VAMP/synaptobrevin and syntaxin , but formed a stable complex ( KD = 1.9 x 10(-7) M ) with P60880 . The minimal essential domain of P60880 required for cleavage by DB00083 involves the segment Met146-Gln197 , and binding was optimal only with full-length P60880 . Proteolysis by BoNT/E required the presence of the domain Ile156-Asp186 . Murine O00161 was cleaved by BoNT/E and , to a reduced extent , by DB00083 , whereas human O00161 was resistant to all clostridial L chains . Lys185Asp or Pro182Arg mutations of human O00161 induced susceptibility toward BoNT/E or toward both DB00083 and BoNT/E , respectively . Crystal structure of Clostridium botulinum neurotoxin protease in a product-bound state : Evidence for noncanonical zinc protease activity . Clostridium botulinum neurotoxins ( BoNTs ) , the most potent toxins known , disrupt neurotransmission through proteolysis of proteins involved in neuroexocytosis . The light chains of BoNTs are unique zinc proteases that have stringent substrate specificity and require exceptionally long substrates . We have determined the crystal structure of the protease domain from BoNT serotype A ( DB00083 ) . The structure reveals a homodimer in a product-bound state , with loop F242-V257 from each monomer deeply buried in its partner 's catalytic site . The loop , which acts as a substrate , is oriented in reverse of the canonical direction for other zinc proteases . The Y249-Y250 peptide bond of the substrate loop is hydrolyzed , leaving the Y249 product carboxylate coordinated to the catalytic zinc . From the crystal structure of the DB00083 protease , detailed models of noncanonical binding and proteolysis can be derived which we propose are also consistent with DB00083 binding and proteolysis of natural substrate synaptosome-associated protein of 25 kDa ( P60880 ) . The proposed DB00083 substrate-binding mode and catalytic mechanism are markedly different from those previously proposed for the BoNT serotype B . Functional characterization of botulinum neurotoxin serotype H as a hybrid of known serotypes F and A ( BoNT F/A ) . A unique strain of Clostridium botulinum ( IBCA10-7060 ) was recently discovered which produces two toxins : botulinum neurotoxin ( BoNT ) serotype B and a novel BoNT reported as serotype H . Previous molecular assessment showed that the light chain ( LC ) of the novel BoNT most resembled the bont of the light chain of known subtype P12259 , while the C-terminus of the heavy chain ( HC ) most resembled the binding domain of serotype A . We evaluated the functionality of both toxins produced in culture by first incorporating an immunoaffinity step using monoclonal antibodies to purify BoNT from culture supernatants and tested each immune-captured neurotoxin with full-length substrates vesicle-associated membrane protein 2 ( P63027 ) , synaptosomal-associated protein 25 ( P60880 ) , syntaxin , and shortened peptides representing the substrates . The BoNT/B produced by this strain behaved as a typical BoNT/B , having immunoaffinity for anti-B monoclonal antibodies and cleaving both full length P63027 and a peptide based on the sequence of P63027 in the expected location . As expected , there was no activity toward P60880 or syntaxin . The novel BoNT demonstrated immunoaffinity for anti-A monoclonal antibodies but did not cleave P60880 as expected for DB00083 . Instead , the novel BoNT cleaved P63027 and P63027 -based peptides in the same location as BoNT/ P12259 . This is the first discovery of a single botulinum neurotoxin with DB00083 antigenicity and BoNT/F light chain function . This work suggests that the newly reported serotype H may actually be a hybrid of previously known BoNT serotype A and serotype F , specifically subtype P12259 . Botulinum toxin 's axonal transport from periphery to the spinal cord . Axonal transport of enzymatically active botulinum toxin A ( DB00083 ) from periphery to the CNS has been described in facial and trigeminal nerve , leading to cleavage of synaptosomal-associated protein 25 ( P60880 ) in central nuclei . Aim of present study was to examine the existence of axonal transport of peripherally applied DB00083 to spinal cord via sciatic nerve . We employed DB00083 -cleaved P60880 immunohistochemistry of lumbar spinal cord after intramuscular and subcutaneous hind limb injections , and intraneural DB00083 sciatic nerve injections . Truncated P60880 in ipsilateral spinal cord ventral horns and dorsal horns appeared after single peripheral DB00083 administrations , even at low intramuscular dose applied ( 5 U/kg ) . Cleaved P60880 appearance in the spinal cord after DB00083 injection into the sciatic nerve was prevented by proximal intrasciatic injection of colchicine ( 5 mM , 2 μl ) . Cleaved P60880 in ventral horn , using choline-acetyltransferase ( P28329 ) double labeling , was localized within cholinergic neurons . These results extend the recent findings on DB00083 retrograde axonal transport in facial and trigeminal nerve . Appearance of truncated P60880 in spinal cord following low-dose peripheral DB00083 suggest that the axonal transport of DB00083 occurs commonly following peripheral application . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Behavioral and immunohistochemical evidence for central antinociceptive activity of botulinum toxin A . Botulinum toxin A ( DB00083 ) is approved for treatment of different cholinergic hyperactivity disorders , and , recently , migraine headache . Although suggested to act only locally , novel observations demonstrated bilateral reduction of pain after unilateral toxin injection , and proposed retrograde axonal transport , presumably in sensory neurons . However , up to now , axonal transport of DB00083 from periphery to CNS was identified only in motoneurons , but with unknown significance . We assessed the effects of low doses of DB00083 injected into the rat whisker pad ( 3.5 U/kg ) or into the sensory trigeminal ganglion ( 1 U/kg ) on formalin-induced facial pain . Axonal transport was prevented by colchicine injection into the trigeminal ganglion ( 5 mM , 2 μl ) . To find the possible site of action of axonally transported DB00083 , we employed immunohistochemical labeling of DB00083 -truncated synaptosomal-associated protein 25 ( P60880 ) in medullary dorsal horn of trigeminal nucleus caudalis after toxin injection into the whisker pad . Both peripheral and intraganglionic DB00083 reduce phase II of formalin-induced pain . Antinociceptive effect of DB00083 was prevented completely by colchicine . DB00083 -truncated P60880 in medullary dorsal horn ( spinal trigeminal nucleus ) was evident 3 days following the peripheral treatment , even with low dose applied ( 3.5 U/kg ) . Presented data provide the first evidence that axonal transport of DB00083 , obligatory for its antinociceptive effects , occurs via sensory neurons and is directed to sensory nociceptive nuclei in the CNS . Botulinum neurotoxin E-insensitive mutants of P60880 fail to bind VAMP but support exocytosis . Neurotransmitter release from synaptic vesicles is mediated by complex machinery , which includes the v- and t- P60880 receptors ( SNAREs ) , vesicle-associated membrane protein ( VAMP ) , synaptotagmin , syntaxin , and synaptosome-associated protein of 25 kDa ( P60880 ) . They are essential for neurotransmitter exocytosis because they are the proteolytic substrates of the clostridial neurotoxins tetanus neurotoxin and botulinum neurotoxins ( BoNTs ) , which cause tetanus and botulism , respectively . Specifically , P60880 is cleaved by both DB00083 and E at separate sites within the COOH-terminus . We now demonstrate , using toxin-insensitive mutants of P60880 , that these two toxins differ in their specificity for the cleavage site . Following modification within the COOH-terminus , the mutants completely resistant to BoNT/E do not bind VAMP but were still able to form a sodium dodecyl sulfate-resistant complex with VAMP and syntaxin . Furthermore , these mutants retain function in vivo , conferring BoNT/E-resistant exocytosis to transfected PC12 cells . These data provide information on structural requirements within the C-terminal domain of P60880 for its function in exocytosis and raise doubts about the significance of in vitro binary interactions for the in vivo functions of synaptic protein complexes . Differential contribution of syntaxin 1 and P60880 to secretion in noradrenergic and adrenergic chromaffin cells . We used botulinum neurotoxins ( BoNT ) to examine whether differences in the secretory activity of noradrenergic and adrenergic chromaffin cells are related to differences in the exocytotic machinery of these two types of bovine adrenal medulla cells . Cleavage of syntaxin and P60880 by BoNT/C1 decreased in a dose-dependent way the release of both noradrenaline and adrenaline , but noradrenaline release was more sensitive to BoNT/C1 . Cleavage of P60880 by DB00083 also had a larger inhibitory effect on noradrenaline release than on adrenaline release . Neither BoNT/C1 nor DB00083 affected the intracellular Ca2+ responses induced by K+-depolarisation , and the extent of the inhibition of K+-evoked catecholamine release by selective blockers of voltage-gated Ca2+ channels was not affected by BoNT/C1 . Therefore , our data do not support the hypothesis of a regulatory effect of syntaxin or P60880 on the activity of Ca2+ channels . The lower sensitivity of adrenaline release to BoNT was not due to a reduced ability of the toxins to enter or to cleave their protein targets in adrenergic cells , since immunoblot analysis showed the cleavage of a larger fraction of syntaxin 1A in adrenergic cells , as compared to the cleavage in noradrenergic cells . The immunoblot analysis also showed larger amounts of syntaxin 1A in noradrenergic chromaffin cells than in adrenergic cells . Thus , in spite of a greater cleavage of syntaxin 1A in adrenergic cells by BoNT/C1 , adrenaline release was less sensitive to BoNT/C1 , suggesting that the release process in noradrenergic cells might be more dependent on syntaxin 1A and P60880 , as compared to adrenergic cells . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Activation of Q8NER1 mediates calcitonin gene-related peptide release , which excites trigeminal sensory neurons and is attenuated by a retargeted botulinum toxin with anti-nociceptive potential . Excessive release of inflammatory/pain mediators from peripheral sensory afferents renders nerve endings hyper-responsive , causing central sensitization and chronic pain . Herein , the basal release of proinflammatory calcitonin gene-related peptide ( P80511 ) was shown to increase the excitability of trigeminal sensory neurons in brainstem slices via CGRP1 receptors because the effect was negated by an antagonist , CGRP8-37 . This excitatory action could be prevented by cleaving synaptosomal-associated protein of M(r) 25,000 ( P60880 ) with botulinum neurotoxin ( BoNT ) type A , a potent inhibitor of exocytosis . Strikingly , DB00083 proved unable to abolish the CGRP1 receptor-mediated effect of capsaicin , a nociceptive Q8NER1 stimulant , or its elevation of P80511 release from trigeminal ganglionic neurons ( TGNs ) in culture . Although the latter was also not susceptible to BoNT/E , apparently attributable to a paucity of its acceptors ( glycosylated synaptic vesicle protein 2 A/B ) , this was overcome by using a recombinant chimera ( EA ) of DB00083 and BoNT/E . It bound effectively to the C isoform of SV2 abundantly expressed in TGNs and cleaved P60880 , indicating that its /A binding domain ( H(C) ) mediated uptake of the active /E protease . The efficacy of /EA is attributable to removal of 26 C-terminal residues from P60880 , precluding formation of SDS-resistant SNARE complexes . In contrast , exocytosis could be evoked after deleting nine of the P60880 residues with /A but only on prolonged elevation of [Ca(2+)](i) with capsaicin . This successful targeting of /EA to nociceptive neurons and inhibition of P80511 release in vitro and in situ highlight its potential as a new therapy for sensory dysmodulation and chronic pain . Dynamin inhibition blocks botulinum neurotoxin type A endocytosis in neurons and delays botulism . The botulinum neurotoxins ( BoNTs ) are di-chain bacterial proteins responsible for the paralytic disease botulism . Following binding to the plasma membrane of cholinergic motor nerve terminals , BoNTs are internalized into an endocytic compartment . Although several endocytic pathways have been characterized in neurons , the molecular mechanism underpinning the uptake of BoNTs at the presynaptic nerve terminal is still unclear . Here , a recombinant DB00083 heavy chain binding domain ( Hc ) was used to unravel the internalization pathway by fluorescence and electron microscopy . DB00083 -Hc initially enters cultured hippocampal neurons in an activity-dependent manner into synaptic vesicles and clathrin-coated vesicles before also entering endosomal structures and multivesicular bodies . We found that inhibiting dynamin with the novel potent Dynasore analog , Dyngo-4a(TM) , was sufficient to abolish DB00083 -Hc internalization and DB00083 -induced P60880 cleavage in hippocampal neurons . Dyngo-4a also interfered with DB00083 -Hc internalization into motor nerve terminals . Furthermore , Dyngo-4a afforded protection against DB00083 -induced paralysis at the rat hemidiaphragm . A significant delay of > 30 % in the onset of botulism was observed in mice injected with Dyngo-4a . Dynamin inhibition therefore provides a therapeutic avenue for the treatment of botulism and other diseases caused by pathogens sharing dynamin-dependent uptake mechanisms . Differential presentation of glutamic acid decarboxylase 65 ( Q05329 ) T cell epitopes among HLA- Q8IUH3 *0401-positive individuals . DB00142 decarboxylase 65 ( Q05329 ) is one of the major autoantigens in type 1 diabetes . We investigated whether there is variation in the processing of Q05329 epitopes between individuals with similar HLA backgrounds and whether the processing characteristics of certain immunogenic epitopes are different in distinct P25054 subpopulations . Using DR401-restricted T cell hybridomas specific for two immunogenic Q05329 epitopes ( 115-127 and 274-286 ) , we demonstrate an epitope-specific presentation pattern in human B-lymphoblastoid cell lines ( B-LCL ) . When pulsed with the Q99259 protein , some Q8IUH3 *0401-positive B-LCL , which presented Q05329 274-286 epitope efficiently , were unable to present the Q05329 115-127 epitope . However , all B-LCL presented synthetic peptides corresponding to either Q99259 epitope . In addition , when pulsed with human serum albumin , all cell lines gave equal stimulation of a DR4-restricted human serum albumin-specific T hybridoma . Q05329 -transfected cell lines displayed the same presentation phenotype , showing that lack of the presentation of the 115-127 epitope was not due to inefficient uptake of the protein . Blood mononuclear adherent cells , B cells , or dendritic cells derived from the same individual displayed the same presentation pattern as observed in B cell lines , suggesting that the defect most likely is genetically determined . Therefore , individual differences in Ag processing may result in the presentation of distinct set of peptides derived from an autoantigen such as Q05329 . This may be an important mechanism for the deviation of the immune response either into a regulatory pathway or into an inflammatory autoimmune reactivity . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Comparison of the catalytic properties of the botulinum neurotoxin subtypes A1 and A5 . Clostridium botulinum neurotoxins ( BoNTs ) cause the life-threatening disease botulism through the inhibition of neurotransmitter release by cleaving essential SNARE proteins . There are seven serologically distinctive types of BoNTs and many subtypes within a serotype have been identified . BoNT/A5 is a recently discovered subtype of type A botulinum neurotoxin which possesses a very high degree of sequence similarity and identity to the well-studied A1 subtype . In the present study , we examined the endopeptidase activity of these two DB00083 subtypes and our results revealed significant differences in substrate binding and cleavage efficiency between subtype A5 and A1 . Distinctive hydrolysis efficiency was observed between the two toxins during cleavage of the native substrate P60880 versus a shortened peptide mimic . N-terminal truncation studies demonstrated that a key region of the P60880 , including the amino acid residues at 151 through 154 located in the remote binding region of the substrate , contributed to the differential catalytic properties between A1 and A5 . Elevated binding affinity of the peptide substrate resulted from including these important residues and enhanced BoNT/A5 's hydrolysis efficiency . In addition , mutations of these amino acid residues affect the proteolytic performance of the two toxins in different ways . This study provides a better understanding of the biological activity of these toxins , their performance characteristics in the Endopep-MS assay to detect BoNT in clinical samples and foods , and is useful for the development of peptide substrates . Peptide inhibitors of botulinum neurotoxin by mRNA display . Botulinum neurotoxins ( BoNTs ) are extremely toxic . The metalloproteases associated with the toxins cleave proteins essential for neurotransmitter secretion . Inhibitors of the metalloprotease are currently sought to control the toxicity of BoNTs . Toward that goal , we produced a synthetic cDNA for the expression and purification of the metalloprotease of DB00083 in Escherichia coli as a biotin-ubiquitin fusion protein , and constructed a combinatorial peptide library to screen for DB00083 light chain inhibitors using mRNA display . A protease assay was developed using immobilized intact P60880 as the substrate . The new peptide inhibitors showed a 10-fold increase in affinity to DB00083 light chain than the parent peptide . Interestingly , the sequences of the new peptide inhibitors showed abundant hydrophobic residues but few hydrophilic residues . The results suggest that mRNA display may provide a general approach in developing peptide inhibitors of BoNTs . Regulation by nitric oxide of endotoxin-induced tissue factor and plasminogen activator inhibitor-1 in endothelial cells . The increase in nitric oxide ( NO ) production in lipopolysaccharide ( LPS ) -induced sepsis is thought to contribute to the development of shock . However , NO could also play an antithrombotic role . Little is known about the modulating effect of NO on the endothelial overexpression and production of tissue factor ( TF ) and plasminogen activator inhibitor-1 ( P05121 ) occurring in endotoxemia . We analyzed the effect of N(G)-nitro-L-arginine-methyl-ester ( L-NAME ) , an inhibitor of NO synthases , and S-nitroso-N-acetyl-D,L-penicillamine ( P60880 ) , a NO donor , on the expression and synthesis of TF and P05121 by LPS-challenged human umbilical vein endothelial cells ( HUVEC ) : L-NAME enhanced the increase in TF mRNA and antigen levels ( P < 0.05 ) observed in LPS-treated HUVEC ; P60880 down-regulated the LPS-induced TF increment ( p < 0.05 ) . However , no effects of NO on regulation of the LPS-dependent increase in P05121 could be seen . Thus , NO could play an antithrombotic role in sepsis by down-regulating the endothelial overexpression and production of TF . Botulinum toxin type A selectivity for certain types of pain is associated with capsaicin-sensitive neurons . Unlike most classical analgesics , botulinum toxin type A ( DB00083 ) does not alter acute nociceptive thresholds , and shows selectivity primarily for allodynic and hyperalgesic responses in certain pain conditions . We hypothesized that this phenomenon might be explained by characterizing the sensory neurons targeted by DB00083 in the central nervous system after its axonal transport . DB00083 's central antinociceptive activity following its application into the rat whisker pad was examined in trigeminal nucleus caudalis ( P24821 ) and higher-level nociceptive brain areas using DB00083 -cleaved synaptosomal-associated protein 25 ( P60880 ) and c-Fos immunohistochemistry . Occurrence of cleaved P60880 in P24821 was examined after nonselective ganglion ablation with formalin or selective denervation of capsaicin-sensitive ( vanilloid receptor-1 or Q8NER1 -expressing ) neurons , and in relation to different cellular and neuronal markers . Regional c-Fos activation and effect of Q8NER1 -expressing afferent denervation on toxin 's antinociceptive action were studied in formalin-induced orofacial pain . DB00083 -cleaved P60880 was observed in P24821 , but not in higher-level nociceptive nuclei . Cleaved P60880 in P24821 disappeared after formalin-induced trigeminal ganglion ablation or capsaicin-induced sensory denervation . Occurrence of cleaved P60880 in P24821 and DB00083 antinociceptive activity in formalin-induced orofacial pain were prevented by denervation with capsaicin . Cleaved P60880 localization demonstrated toxin 's presynaptic activity in Q8NER1 -expressing neurons . DB00083 reduced the c-Fos activation in P24821 , locus coeruleus , and periaqueductal gray . Present experiments suggest that DB00083 alters the nociceptive transmission at the central synapse of primary afferents . Targeting of Q8NER1 -expressing neurons might be associated with observed selectivity of DB00083 action only in certain types of pain . Gene expression analysis of spontaneously hypertensive rat cerebral cortex following transient focal cerebral ischemia . Identification of novel modulators of ischemic neuronal death helps in developing new strategies to prevent the stroke-induced neurological dysfunction . Hence , the present study evaluated the gene expression changes in rat cerebral cortex at 6 and 24 h of reperfusion following transient middle cerebral artery occlusion ( MCAO ) by GeneChip analysis . Transient MCAO resulted in selective increased mRNA levels of genes involved in stress , inflammation , transcription and plasticity , and decreased mRNA levels of genes which control neurotransmitter function and ionic balance . In addition to a number of established ischemia-related genes , many genes not previously implicated in transient focal ischemia-induced brain damage [ suppressor of cytokine signaling ( Q9NSE2 ) -3 , DB02527 responsive element modulator ( Q03060 ) , cytosolic retinol binding protein ( P09455 ) , silencer factor-B , survival motor neuron ( SMN ) , interferon-gamma regulatory factor-1 ( P10914 ) , galanin , neurotrimin , proteasome subunit RC8 , synaptosomal-associated protein ( P60880 ) -25 A and B , synapsin 1a , neurexin 1-beta , ras-related rab3 , vesicular GABA transporter ( Q9H598 ) , digoxin carrier protein , neuronal calcium sensor-1 and neurodap ] were observed to be altered in the ischemic cortex . Real-time PCR confirmed the GeneChip results for several of these transcripts . O14543 is a gene up-regulated after ischemia which modulates inflammation by controlling cytokine levels . Antisense knockdown of ischemia-induced O14543 protein expression exacerbated transient MCAO-induced infarct volume assigning a neuroprotective role to O14543 , a gene not heretofore implicated in ischemic neuronal damage . Traffic of botulinum toxins A and E in excitatory and inhibitory neurons . Botulinum neurotoxins ( BoNTs ) , proteases specific for the SNARE proteins , are used to study the molecular machinery supporting exocytosis and are used to treat human diseases characterized by cholinergic hyperactivity . The recent extension of the use of BoNTs to central nervous system ( CNS ) pathologies prompted the study of their traffic in central neurons . We used fluorescent DB00083 and BoNT/E to study the penetration , the translocation and the catalytic action of these toxins in excitatory and inhibitory neurons . We show that DB00083 and BoNT/E , besides preferentially inhibiting synaptic vesicle recycling at glutamatergic relative to Gamma-aminobutyric acid ( GABA ) -ergic neurons , are more efficient in impairing the release of excitatory than inhibitory neurotransmitter from brain synaptosomes . This differential effect does not result from a defective penetration of the toxin in line with the presence of the DB00083 receptor , synaptic vesicle protein 2 ( SV2 ) , in both types of neurons . Interestingly , exogenous expression of P60880 in GABAergic neurons confers sensitivity to DB00083 . These results indicate that the expression of the toxin substrate , and not the toxin penetration , most likely accounts for the distinct effects of the two neurotoxins at the two types of terminals and support the use of BoNTs for the therapy of CNS diseases caused by the altered activity of selected neuronal populations . Inhibition of neurotransmitter release by clostridial neurotoxins correlates with specific proteolysis of synaptosomal proteins . Rat brain synaptosomes were used to study the effect of several clostridial neurotoxins on the neurotransmitter release . In this system the blockade of transmitter release correlated with the proteolytic activity of the toxins . Blockade of glutamate release was linked to selective proteolysis of one of the following synaptic proteins : synaptobrevin ( BoNT/D , BoNT/F ) ; P60880 ( DB00083 , BoNT/E ) , or HPC-1/syntaxin ( BoNT/C1 ) . All the toxins used had an inhibitory effect on synaptosomes with the exception of BoNT/F . BoNT/F cleaved synaptobrevin in permeabilized synaptosomes but failed to produce the same effect on intact synaptosomes . DB00435 induces apoptosis in GM- P04141 -treated eosinophils via caspase-6-dependent lamin and DNA fragmentation . Asthma is characterized by accumulation of eosinophils in the lungs and delayed apoptosis may be one mechanism leading to eosinophilia . DB00435 ( NO ) , present in inflamed lungs , has been shown to possess both anti- and proeosinophilic properties . We previously showed that NO induces apoptosis in the presence of survival prolonging cytokine P05113 in human eosinophils . In the present study , we examined the intracellular mechanisms of NO-induced apoptosis in granulocyte macrophage-colony stimulating factor ( GM- P04141 ) -treated eosinophils concentrating on the role of caspases and calpains . Eosinophils were isolated from human blood and apoptosis was determined by relative DNA fragmentation assay , morphological analysis and/or Annexin-V FITC assay . We showed that NO-donor S-nitroso-N-acetyl-d,l-penicillamine ( P60880 ) induced apoptosis in GM- P04141 -treated eosinophils . P60880 -induced DNA fragmentation was totally prevented by an inhibitor of caspase-6 ( Z-VEID-FMK ) . Decreased levels of caspase-6 proenzyme and increased amounts of cleaved lamin A/C in P60880 -treated cells indicated activation of caspase-6 . Furthermore , P60880 -induced lamin A/C and B fragmentation was totally abolished by an inhibitor of caspase-6 . According to our results , caspase-6 mediates lamin and DNA fragmentation also in spontaneously dying eosinophils . Inhibitor of calpains prevented most of DNA fragmentation related to spontaneous apoptosis but had no effect in eosinophils undergoing NO-induced apoptosis . In the present study we showed that caspase-6 is essential for the executive phase involving lamin and DNA fragmentation in both NO-induced and spontaneous eosinophil apoptosis . However , differences in the involvement of calpains suggest that the intracellular signalling in NO-induced apoptosis has specific features at the level of proteases . This study demonstrates new mechanisms for NO-induced and spontaneous apoptosis in human eosinophils . Ca(2+) influx and DB02527 elevation overcame botulinum toxin A but not tetanus toxin inhibition of insulin exocytosis . Previous reports showed that cleavage of vesicle-associated membrane protein-2 ( P63027 ) and synaptosomal-associated protein of 25 kDa ( P60880 ) by clostridial neurotoxins in permeabilized insulin-secreting beta-cells inhibited Ca(2+)-evoked insulin secretion . In these reports , the soluble N-ethylmaleimide-sensitive factor attachment protein target receptor proteins might have formed complexes , which preclude full accessibility of the putative sites for neurotoxin cleavage . In this work , P63027 and P60880 were effectively cleaved before they formed toxin-insensitive complexes by transient transfection of insulinoma HIT or P01308 -1 cells with tetanus toxin ( TeTx ) or botulinum neurotoxin A ( DB00083 ) , as shown by immunoblotting and immunofluorescence microscopy . This resulted in an inhibition of Ca(2+) ( glucose or DB00761 ) -evoked insulin release proportionate to the transfection efficiency ( 40-50 % ) and an accumulation of insulin granules . With the use of patch-clamp capacitance measurements , Ca(2+)-evoked exocytosis by membrane depolarization to -10 mV was abolished by TeTx ( 6 % of control ) but only moderately inhibited by DB00083 ( 30 % of control ) . Depolarization to 0 mV to maximize Ca(2+) influx partially overcame DB00083 ( 50 % of control ) but not TeTx inhibition . Of note , DB02527 activation potentiated Ca(2+)-evoked secretion by 129 % in control cells but only 55 % in DB00083 -transfected cells and had negligible effects in TeTx-transfected cells . These results indicate that , whereas P63027 is absolutely necessary for insulin exocytosis , the effects of P60880 depletion on exocytosis , perhaps on insulin granule pool priming or mobilization steps , could be partially reversed by higher levels of Ca(2+) or DB02527 potentiation . Recombinant holotoxoid vaccine against botulism . The botulinum neurotoxins ( BoNT ) are the most toxic proteins for humans and designated " Category A Select Agents. " The current vaccine against botulism is in limited supply , and there is a need to develop new vaccine strategies . A recombinant DB00083 toxoid was produced in Clostridium botulinum that contained a double amino acid substitution , R363A Y365F ( termed DB00083 (RYM) ) . DB00083 (RYM) was noncatalytic for P60880 and nontoxic for mice . Immunization with DB00083 (RYM) protected mice from challenge at levels that were similar to chemically inactivated DB00083 toxoid . DB00083 (RYM) elicited an immune response against the light-chain and heavy-chain components of the toxin . Neutralizing anti- DB00083 (RYM) sera blocked BoNT toxicity in primary cortical neurons and blocked ganglioside binding by the heavy chain . DB00083 (RYM) represents a viable vaccine candidate for a holotoxoid against botulism . Hypersensitive detection and quantitation of DB00083 by IgY antibody against substrate linear-peptide . Botulinum neurotoxin A ( DB00083 ) , the most acutely poisonous substance to humans known , cleave its P60880 substrate with high specificity . Based on the endopeptidase activity , different methods have been developed to detect DB00083 , but most lack ideal reproducibility or sensitivity , or suffer from long-term or unwanted interferences . In this study , we developed a simple method to detect and quantitate trace amounts of botulinum neurotoxin A using the IgY antibody against a linear-peptide substrate . The effects of reaction buffer , time , and temperature were analyzed and optimized . When the optimized assay was used to detect DB00083 , the limit of detection of the assay was 0.01 mouse LD50 ( 0.04 pg ) , and the limit of quantitation was 0.12 mouse LD50/ml ( 0.48 pg ) . The findings also showed favorable specificity of detecting DB00083 . When used to detect DB00083 in milk or human serum , the proposed assay exhibited good quantitative accuracy ( 88 % < recovery < 111 % ; inter- and intra-assay CVs < 18 % ) . This method of detection took less than 3 h to complete , indicating that it can be a valuable method of detecting DB00083 in food or clinical diagnosis . Comparison of fluorigenic peptide substrates PL50 , SNAPTide , and BoTest A/E for DB00083 detection and quantification : exosite binding confers high-assay sensitivity . Detection and quantification of low doses of botulinum toxin serotype A ( DB00083 ) in medicinal preparations require precise and sensitive methods . With mounting pressure from governmental authorities to replace the mouse LD50 assay , interest in alternative methods such as the endopeptidase assay , quantifying the toxin active moiety , is growing . Using internal collision-induced fluorescence quenching , Pharmaleads produced peptides encompassing the P60880 cleavage site : a 17-mer ( PL63 ) and a 48-mer ( PL50 ) reaching the previously identified α-exosite , with PL50 showing higher apparent affinity for DB00083 . Peptide mapping experiments revealed that this increased affinity is mainly due to a connecting peptide sequence between the N-terminus of PL63 and the α-exosite , identifying a new cooperative exosite on DB00083 . Other endopeptidase substrates available , including SNAPTide and BoTest A/E , are both based on fluorescent resonance energy transfer ( FRET ) technology . To compare these assays , their limits of detection and quantification were determined using light chain or 150-kDa DB00083 . Detection limits of PL50 and BoTest were over 100 times better than those using SNAPtide in standard conditions . Although the BoTest possessed a detection limit around 0.2 pM for either DB00083 form , its quantification limit ( 9.7 pM ) using purified DB00083 was 12 times inferior to PL50 , estimated at 0.8 pM , suitable for medicinal preparation quantification . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Disruption of lipid rafts enhances activity of botulinum neurotoxin serotype A . Botulinum neurotoxin serotype A ( DB00083 ) , one of seven serotypes of botulinum neurotoxin , is taken up by neurons of the peripheral nervous system . Within the neurons it catalyzes cleavage of the synaptosomal-associated protein having a mass of 25kDa , P60880 , thereby blocking neurotransmission . DB00083 has been shown to interact with SV2 , as well as gangliosides that are often found in lipid rafts . Lipid rafts are microdomains that can be found on the outer leaflet of the plasma membrane and are enriched in cholesterol and glycosphingolipids . To determine whether lipid rafts are needed for DB00083 activity , those associated with the plasma membranes of murine N2a neuroblastoma cells were disrupted using either methyl-beta-cyclodextrin or filipin . Disruption of cholesterol-containing lipid rafts by either reagent did not prevent the action of DB00083 on N2a cells , in fact activity was enhanced . While our results indicate that disruption of lipid rafts enhances DB00083 activity , disruption of clathrin-dependent endocytosis appeared to be inhibitory . Capillary electrophoresis laser-induced fluorescence for screening combinatorial peptide libraries in assays of botulinum neurotoxin A . Botulinum neurotoxin serotype A ( DB00083 ) is a proteolytic enzyme that induces muscle paralysis . It is a cause of food poisoning , a potential bioterrorist threat and , in low doses an emerging pharmaceutical product . No effective treatment is currently available for BoNT intoxication . Previously we developed a DB00083 light chain enzyme assay using a peptide substrate based on the P60880 protein target , with HPLC separation and UV detection of assay products , and applied the method to screen combinatorial peptide libraries for inhibitory activity to DB00083 . We now report on development of a capillary electrophoresis laser-induced fluorescence ( CE- P15018 ) method for measuring DB00083 activity . The enzyme assay products were labeled with CBQCA dye followed by CE separation on a bare fused silica column in a HEPES-based buffer and P15018 detection . All assay products were separated in CE within 8 min compared to incomplete separation of assay products within 1h by HPLC . The labeled products showed linear dependence of intensity versus concentration , and quantitative mole-fraction assignments . We used the CE- P15018 method to screen combinatorial peptide libraries for potential modulating effects on DB00083 peptidase activity . With some of the libraries , peptides co-migrated with assay products and interfered with quantitation . In such cases , interference was reduced by substituting sodium dodecyl sulfate ( SDS ) for Tween-20 in the running buffer . Separation in the capillaries then occurred by micellar electrokinetic chromatography ( MEKC ) . The CE- P15018 method is quick and lends itself to high-throughput or microfluidic formats . Neutralization of botulinum neurotoxin by a human monoclonal antibody specific for the catalytic light chain . BACKGROUND : Botulinum neurotoxins ( BoNT ) are a family of category A select bioterror agents and the most potent biological toxins known . Cloned antibody therapeutics hold considerable promise as BoNT therapeutics , but the therapeutic utility of antibodies that bind the BoNT light chain domain ( LC ) , a metalloprotease that functions in the cytosol of cholinergic neurons , has not been thoroughly explored . METHODS AND FINDINGS : We used an optimized hybridoma method to clone a fully human antibody specific for the LC of serotype A BoNT ( DB00083 ) . The 4LCA antibody demonstrated potent in vivo neutralization when administered alone and collaborated with an antibody specific for the HC . In Neuro-2a neuroblastoma cells , the 4LCA antibody prevented the cleavage of the DB00083 proteolytic target , P60880 . Unlike an antibody specific for the HC , the 4LCA antibody did not block entry of DB00083 into cultured cells . Instead , it was taken up into synaptic vesicles along with DB00083 . The 4LCA antibody also directly inhibited DB00083 catalytic activity in vitro . CONCLUSIONS : An antibody specific for the DB00083 LC can potently inhibit DB00083 in vivo and in vitro , using mechanisms not previously associated with BoNT-neutralizing antibodies . Antibodies specific for BoNT LC may be valuable components of an antibody antidote for BoNT exposure . Expression and purification of the light chain of botulinum neurotoxin A : a single mutation abolishes its cleavage of P60880 and neurotoxicity after reconstitution with the heavy chain . Botulinum neurotoxin type A ( DB00083 ) selectively and irreversibly inhibits acetylcholine release from peripheral nerve endings . While the toxin 's heavy ( H ) chain contributes to neuronal binding and internalization , its light ( L ) chain is a Zn(2+)-dependent endoprotease that intracellularly cleaves synaptosomal-associated protein of M(r) = 25 kDa ( P60880 ) . For research and clinical exploitation of this uniquely-acting neurotoxin , recombinant wild-type L chain was produced together with a mutant in which His227 in the Zn(2+)-binding motif was substituted by DB00135 . The PCR-amplified wild-type and mutant L chain genes were cloned , fused to the gene for maltose-binding protein , and expressed at high levels in Escherichia coli . The soluble fusion proteins were purified using amylose affinity chromatography , and , after factor Xa cleavage , the free L chains were isolated . The wild-type was shown to proteolyze P60880 at a rate approaching that of the native chain while the mutant was inactive . Reconstitution of the pure wild-type L chain with native homogeneous H chain yielded a disulfide-linked dichain form that inhibited neuromuscular transmission in vitro and produced the symptoms of botulism in vivo . After reconstitution with the H chain , the Tyr227 mutant L chain failed to show any neuroparalytic activity in either of these assays . This methodology allows , for the first time , routine preparation of recombinant forms of the L chain that are needed to decipher the molecular details of its interaction with substrate and , thereby , assist the design of effective inhibitors. ( ABSTRACT TRUNCATED AT 250 WORDS ) Stable expression of a neuronal dopaminergic progenitor phenotype in cell lines derived from human amniotic fluid cells . Cells from human amniotic fluid derived from the fetus are considered a source of multipotent cells . Their properties have not been fully exploited , partially because unlike other embryonic sources such as embryonic stem ( ES ) cells , cell lines from amniocentesis samples have not been generated . We have established and characterized the properties of eight individual cell lines . Flow cytometry using several cell surface markers showed that all cell lines generated consisted of homogeneous populations that lack HLAII antigenicity . Using a combination of immunocytochemistry , Western blotting , and RT-PCR , we found weak expression of Oct4 and nestin and strong expression of tubulin-betaIII , P11137 , and tau . Specific markers for cholinergic , (nor)adrenergic , and GABAergic neurons or glia were weakly expressed or absent , whereas expression of factors implicated in early induction of dopaminergic neurons , TGF-beta3 and beta-catenin were present . Further analysis showed strong expression of EN-1 , c- P07949 , PTX3 , and P43354 essential for induction and survival of midbrain dopaminergic neurons , TH , P20711 , and Q05940 components of dopamine synthesis and secretion , and syntaxin1A and P60880 necessary for neurotransmitter exocytosis . This phenotype was retained throughout passages and up to the current passage 36 . Expression of neuronal and dopaminergic markers in individual AF cell lines was comparable to expression in neurons induced from ES cells and in IMR-32 and SH-SY5Y neuroblastomas . Our data show that cell lines can be derived from subcultures of amniocentesis , and are primarily composed of a population of progenitors with a phenotype similar to that of committed mesencephalic dopaminergic neurons . Comparative role of neurotoxin-associated proteins in the structural stability and endopeptidase activity of botulinum neurotoxin complex types A and E . Seven serotypes of botulinum neurotoxins , the most toxic substances known to mankind , are each produced by different strains of Clostridium botulinum along with a group of neurotoxin-associated proteins ( NAPs ) . NAPs play a critical role in the toxicoinfection process of botulism in addition to their role in protecting the neurotoxin from proteolytic digestion in the GI tract as well as from adverse environmental conditions . In this study we have investigated the effect of temperature on the structural and functional stability of DB00083 complex ( BoNT/AC ) and BoNT/E complex ( BoNT/EC ) . Although the NAPs in the two complexes are quite different , both groups of NAPs activate the endopeptidase activities of their BoNTs without any need to reduce the disulfide bonds between light and heavy chains of respective BoNTs . BoNT/AC attains optimum enzyme activity at the physiological temperature of 37 degrees C whereas BoNT/EC is maximally active at 45 degrees C , and this is accompanied by conformational alterations in its polypeptide folding at this temperature , leading to favorable binding with its intracellular substrate , P60880 , and subsequent cleavage of the latter . DB00083 in its complex form is found to be structurally more stable against temperature whereas BoNT/E in its complex form is functionally better protected against temperature . Based on the analysis of isolated NAPs we have observed that the structural stability of the BoNT/AC is contributed by the NAPs . In addition to the unique structural conditions in which the enzyme remains active , functional stability of botulinum neurotoxins against temperature plays a critical role in the survival of the agent in cooked food and in food-borne botulism . [ Changes in chemokine receptor 4 , interleukin-6 , and collagen X expression in the ATDC5 cell line stimulated by cyclic tensile strain and stromal cell-derived factor-1 ] . OBJECTIVE : This study further explores the stromal cell-derived factor-1 ( P48061 ) /chemokine receptor 4 ( P61073 ) signaling axis mechanism in temporomandibular joint osteoarthritis ( OA ) by detecting the changes in P61073 , interleukin ( IL ) -6 , and collagen X expression in the ATDC5 cell line stimulated by the cyclic tensile strain and P48061 . METHODS : P01308 -transferrin-selenium ( ITS ) was used to induce ATDC5 cells to differentiate into chondrocyte-like cells . After three weeks , the cells were divided into two groups : those with and without cyclic tensile strain . These groups were further divided into the negative control and P48061 groups . Strain force of 20 % was applied . After 12 h , the total proteins were extracted from cells of the four groups , and Western blot analysis was used to detect the changes in P61073 , P05231 , and collagen X expression . RESULTS : P48061 could enhance P61073 , P05231 , and collagen X expressions in the chondrocytes , and 20 % tensile strain force could further upregulate the three factors . CONCLUSION : Under abnormal tensile force , P48061 can upregulate its specific receptor P61073 , thus increasing its-binding efficiency and resulting in the activation of the P48061 / P61073 axis . This condition enhances the expressions of P05231 and other inflammatory factors and directly damages to cartilage tissue . Such damage directly promotes chondrocyte hypertrophy , which enhances collagen X expression . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . S,S'-dinitrosobucillamine , a new nitric oxide donor , induces a better vasorelaxation than other S-nitrosothiols . S-nitrosothiols ( RSNO ) are considered as potential drugs for delivering nitric oxide ( NO ) or related species in cardiovascular disorders associated with decrease in NO bioavailability . We have synthesized a new RSNO , i.e. S,S'-dinitrosobucillamine ( BUC(NO)2 ) , which combines in its structure two S-mononitrosothiols , S-nitroso-N-acetylpenicillamine ( P60880 ) and S-nitroso- DB06151 ( NACNO ) . Synthesized BUC(NO)2 was structurally characterized using high-performance liquid chromatography/mass spectrometry ( HPLC/MS ) , (1)H nuclear magnetic resonance ( (1)H NMR ) , infrared ( IR ) and UV-visible spectroscopies , and thermal analysis ; resulting data are consistent with the expected structure . The vasorelaxant effect of BUC(NO)2 was evaluated using isolated rat aortic rings and compared to P60880 , NACNO , and to an equimolar mixture of NACNO plus P60880 in order to mimic the number of NO contained in a BUC(NO)2 molecule . BUC(NO)2 ( pD2=7.8±0.1 ) was more potent in vasorelaxation than NACNO ( pD2=6.4±0.2 ) , P60880 ( pD2=6.7±0.1 ) and the mixture of P60880 plus NACNO ( pD2=6.7±0.2 ) . The release of NO from BUC(NO)2 was 6-fold that of the basal value and significantly higher than the release of NO from the P60880 plus NACNO mixture ( 4-fold increase versus basal value ) . Finally , the role of protein disulfide isomerase ( P07237 ) in BUC(NO)2 metabolism was investigated . Vasorelaxant effect ( pD2=6.8±0.2 ) and NO release decreased in the presence of a P07237 inhibitor ( both P < 0.05 versus BUC(NO)2 ) . In conclusion , BUC(NO)2 releases a larger amount of NO into the aorta , partially through P07237 activation , and induces vasorelaxation at lower concentrations than other RSNO previously reported . Molecular targets of botulinum toxin at the mammalian neuromuscular junction . The molecular targets of botulinum neurotoxins ( BoNTs ) are SNARE ( soluble N-ethylmaleimide-sensitive factor-attachment protein-receptor ) proteins necessary for neurotransmitter release . BoNT are powerful therapeutic agents in the treatment of numerous neurological disorders . The goals of this study were to ( 1 ) assess toxin diffusion by measuring substrate cleavage in adjacent and distant muscles , and ( 2 ) characterize the clinical course using SNARE protein chemistry . A small volume of DB00083 was injected unilaterally into the mouse gastrocnemius muscle . Motor impairment was limited to the toxin-treated limb . No systemic illness or deaths occurred . At five time points , a subset of mice were killed , and muscles from both hindlimbs , and the diaphragm , were collected . Protein samples were examined for changes in P60880 ( synaptosomal-associated protein of Mr = 25 kDa ) using immunochemistry . P60880 cleavage product was noted in the toxin-treated limb as early as 1 day postinjection and continued through day 28 . Onset and peak levels of substrate cleavage corresponded to the onset and peak clinical response . Cleavage was observed in adjacent and distant muscles , demonstrating that substrate cleavage is a sensitive indicator of toxin diffusion . Significant increases in full-length P60880 and vesicle-associated membrane protein II were evident early in the impaired limb and continued through day 28 . The increased SNARE protein most likely originates from nerve terminal sprouts .

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interacts_with DB01101?

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Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Molecular characterisation of a panel of human ovarian carcinoma xenografts . In a panel of 16 human ovarian tumours transplanted in nude mice , the expression of genes involved in cell cycle regulation and in response to drug treatment were characterised . In the 16 tumours analysed we could not detect overexpression of Erb-B2 oncogene while expression of P08183 mRNA was not detected in 11/15 samples and was low in 4/15 tumours . Only three tumours had mutations in the p53 gene exons 5-8 and one of these mutations did not result in any amino acid alteration . The levels of mRNA for cyclins A , D1 and E were heterogeneous with some tumours expressing high levels and others not expressing them at all . The same was found for the cyclin dependent kinases ( CDK ) P24941 and P11802 and for CDK inhibitors P38936 / P38936 , p27/ P46527 and p16/CDKN2 . Two genes belonging to the nucleotide excision repair , P07992 and P19447 were detectable in all the samples examined , as were the genes P16455 and P20916 , also involved in DNA repair . The data indicate a heterogeneity in the expression of genes considered to be involved in the cellular responses to cytotoxic drug treatment and indicate the possibility of using these tumour models to test specifically molecules with a defined mechanism of action . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Phase II study of weekly intravenous oxaliplatin combined with oral daily capecitabine and radiotherapy with biologic correlates in neoadjuvant treatment of rectal adenocarcinoma . PURPOSE : To evaluate the efficacy of a combination of capecitabine , oxaliplatin , and radiotherapy ( RT ) in the neoadjuvant treatment of Stage II and III rectal cancers . METHODS : DB01101 was given at 725 mg/m(2) orally twice daily Monday through Friday concurrently with RT . DB00526 was given intravenously at 50 mg/m(2) once weekly five times starting the first day of RT . The radiation dose was 50.4 Gy in 28 fractions ( 1.8 Gy/fraction ) , five fractions weekly . Endorectal tumor biopsies were obtained before treatment and on the third day of treatment to explore the effects of treatment on thymidine phosphorylase , thymidylate synthase , excision repair cross-complementing rodent repair deficiency complementation group 1 ( P07992 ) , and apoptosis . RESULTS : A total of 25 patients were enrolled in this study ; 6 patients ( 24 % ) had a complete pathologic response . T-downstaging occurred in 52 % of patients , and N-downstaging occurred in 53 % . Grade 3 diarrhea was the most common Grade 3-4 toxicity , occurring in 20 % of patients . Only 2 patients experienced disease recurrence , with a median of 20 months of follow-up . P04818 , thymidine phosphorylase , P07992 , and apoptosis did not vary significantly between the pretreatment and Day 3 tumor biopsies , nor did they predict for T-downstaging or a complete pathologic response . CONCLUSION : DB01101 at 725 mg/m(2) orally twice daily , oxaliplatin 50 mg/m(2)/wk , and RT at 50.4 Gy is an effective neoadjuvant combination for Stage II and III rectal cancer and results in a greater rate of complete pathologic responses than historically shown in fluoropyrimidine plus RT controls . Overexpression of the two nucleotide excision repair genes P07992 and Q01831 in human hepatocellular carcinoma . BACKGROUND/AIMS : Little is known about the nucleotide excision repair ( P55055 ) pathway in the resistance of human hepatocellular carcinoma ( HCC ) to chemotherapeutics . We investigated expression of several P55055 genes in human HCC and matching non-tumor tissue ( NT ) and in normal liver . METHODS : Expression of Q13216 , Q03468 , Q01831 , P54727 , P23025 , P19447 , P07992 and p53 genes was analyzed by quantitative RT-PCR and immunoblotting in 26 HCC and 9 normal livers . RESULTS : The seven P55055 genes and p53 were frequently overexpressed in HCC compared to matched NT . P23025 , Q01831 , P54727 and P07992 mRNA levels were significantly increased ( p < 0.05 ) in HCC arising in cirrhotic livers compared to non fibrotic tissue . Moreover , expression of P07992 , P23025 and Q01831 mRNA was significantly augmented in HCC , even more in tumors arising in cirrhotic liver . P07992 , Q01831 ad P23025 mRNA levels were highly correlated in NT and HCC . Q01831 and P07992 protein levels were also increased in HCC . CONCLUSIONS : Our findings strongly suggest that overexpression of two key genes involved in the early steps of the P55055 process , P07992 and Q01831 , is associated with liver fibrogenesis and cancer and could be related to the well recognized resistance of HCC to chemotherapeutics . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Analysis of over 10,000 Cases finds no association between previously reported candidate polymorphisms and ovarian cancer outcome . BACKGROUND : Ovarian cancer is a leading cause of cancer-related death among women . In an effort to understand contributors to disease outcome , we evaluated single-nucleotide polymorphisms ( SNP ) previously associated with ovarian cancer recurrence or survival , specifically in angiogenesis , inflammation , mitosis , and drug disposition genes . METHODS : Twenty-seven SNPs in P40337 , P14210 , Q14116 , P22694 , P08183 , P10632 , P18074 , and P07992 previously associated with ovarian cancer outcome were genotyped in 10,084 invasive cases from 28 studies from the Ovarian Cancer Association Consortium with over 37,000-observed person-years and 4,478 deaths . Cox proportional hazards models were used to examine the association between candidate SNPs and ovarian cancer recurrence or survival with and without adjustment for key covariates . RESULTS : We observed no association between genotype and ovarian cancer recurrence or survival for any of the SNPs examined . CONCLUSIONS : These results refute prior associations between these SNPs and ovarian cancer outcome and underscore the importance of maximally powered genetic association studies . Q9P2X3 : These variants should not be used in prognostic models . Alternate approaches to uncovering inherited prognostic factors , if they exist , are needed . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . The molecular basis of the chemosensitivity of metastatic cutaneous melanoma to chemotherapy . BACKGROUND : Chemotherapy benefits relatively few patients with cutaneous melanoma . The assessment of tumour chemosensitivity by the DB00171 -based tumour chemosensitivity assay ( DB00171 -TCA ) has shown strong correlation with outcome in cutaneous melanoma , but requires fresh tissue and dedicated laboratory facilities . AIM : To examine whether the results of the DB00171 -TCA correlate with the expression of genes known to be involved in resistance to chemotherapy , based on the hypothesis that the molecular basis of chemosensitivity lies within known drug resistance mechanisms . METHOD : The chemosensitivity of 47 cutaneous melanomas was assessed using the DB00171 -TCA and correlated with quantitative expression of 93 resistance genes measured by quantitative reverse transcriptase PCR ( qRT-PCR ) in a Taqman Array after extraction of total RNA from formalin-fixed paraffin-embedded tissue . RESULTS : Drugs susceptible to particular resistance mechanisms showed good correlation with genes linked to these mechanisms using signatures of up to 17 genes . Comparison of these signatures for DTIC , treosulfan and cisplatin showed several genes in common . HSP70 , at least one human epidermal growth factor receptor , genes involved in apoptosis ( IAP2 , P60484 ) and DNA repair ( P07992 , P23025 , P18887 , P12956 ) were present for these agents , as well as genes involved in the regulation of proliferation ( Ki67 , P38936 , p27 ) . The combinations tested included genes represented in the single agent signatures . CONCLUSIONS : These data suggest that melanoma chemosensitivity is influenced by known resistance mechanisms , including susceptibility to apoptosis . Use of a candidate gene approach may increase understanding of the mechanisms underlying chemosensitivity to drugs active against melanoma and provide signatures with predictive value . FLEX data , P01116 and P07992 testing in oncology . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . Dual inhibition of P00533 and VEGFR pathways in combination with irradiation : antitumour supra-additive effects on human head and neck cancer xenografts . The aim of this study was to investigate the effects of combining antiangiogenic treatment , epidermal growth factor receptor ( P00533 ) targeting and irradiation ( RT ) . We evaluated DB04849 , a highly potent , orally active , vascular endothelial growth factor ( P15692 ) signalling inhibitor , gefitinib , an P00533 tyrosine kinase inhibitor and RT . The antitumour efficacy of these treatments , administered alone and in combination for 2 weeks , was assessed in a P15692 -secreting human head and neck tumour cell line , CAL33 that highly expresses P00533 , established as xenografts ( 250 mm(3) ) in nude mice . The median time to reach a tumour volume of 1000 mm(3) was significantly increased for DB04849 or gefitinib alone compared with the control . Greater inhibition of tumour growth was seen with the combination of DB04849 +gefitinib compared with either drug alone , and the triple combination compared with either DB04849 +gefitinib or RT alone . The intensity of endothelial cell staining was slightly reduced by each agent given alone , and markedly diminished by the double or triple combination . The triple combination almost completely abolished cell proliferation . The marked RT-induced enhancement in the DNA-repair enzyme P07992 expression was totally abolished by the triple combination . This observation could help to explain the supra-additive antitumour effect produced by this combination and could provide a basis for future innovative clinical trials . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Specific Biomarkers Are Associated with Docetaxeland Gemcitabine-Resistant NSCLC Cell Lines . Five-year survival rate for lung cancer is limited to 10 % to 15 % . Therefore , the identification of novel therapeutic prognostic factors is an urgent requirement . The aim of this study is thus to highlight specific biomarkers in chemoresistant non-small cell lung cancer cell lines . Therefore , we checked-in the control condition as well as after short-term pharmacological treatment with either docetaxel or gemcitabine-the expression of genes such as tumor suppressor genes ( CDKN2A , P53355 , P49789 , P09211 , P16455 , RARβ2 , RASSF1A , and P35625 ) , genes associated with drug resistance ( P38398 , P35354 , P07992 , P17936 , P23921 , and Q13509 ) , and stemness-related genes ( CD133 , Q01860 , and O43623 ) in two cellular models of squamous carcinoma ( CAEP ) and adenocarcinoma ( RAL ) of the lung originally established . Their promoter methylation profile was also evaluated . Drug-related genes were upregulated . DB00515 resistance matched with high levels of P38398 and P07992 in both cell lines ; docetaxel sensitivity of CAEP cells was associated to levels of Q13509 lower than RAL cells . Although CAEP cells were more sensitive to gemcitabine , both cell lines showed high levels of P23921 . Stemness-related genes were downregulated in the control condition but became upregulated in docetaxel-resistant cells , indicating the selection of a population with stemness features . We did not find an unequivocal correspondence between gene expression and respective DNA promoter methylation status , suggesting the involvement of additional mechanisms of gene expression regulation . These results highlight specific biomarkers consistent with the different responses of the two cell lines to standard pharmacological treatments and indicate specific molecular traits for their chemoresistance . Stress-related gene expression in mice treated with inorganic arsenicals . Arsenic ( As ) is an environmental chemical of high concern for human health . Acute toxicity of arsenic is dependent on its chemical forms and proximity to high local arsenic concentrations is one of the mechanisms for cell death . This study was designed to define acute arsenic-induced stress-related gene expression in vivo . Mice were injected sc with either sodium arsenite [ As(III) , 100 micromol/kg ] , sodium arsenate [ As(V) , 300 micromol/kg ] , or saline . To examine stress-related gene expression , livers were removed 3 h after arsenic injection for RNA and protein extraction . The Atlas Mouse Stress/Toxicology array revealed that the expression of genes related to stress , DNA damage , and metabolism was altered by acute arsenic treatments . Expression of heme oxygenase 1 ( P09601 ) , a hallmark for arsenic-induced stress , was increased 10-fold , along with increases in heat shock protein-60 ( HSP60 ) , DNA damage inducible protein P24522 , and the DNA excision repair protein P07992 . Downregulation of certain cytochrome P450 enzymes occurred with arsenic treatment . Multiprobe RNase protection assay revealed the activation of the c-Jun/AP-1 transcription complex after arsenic treatments . Western blot analysis further confirmed the enhanced production of arsenic-induced stress proteins such as P09601 , HSP70 , HSP90 , metallothionein , the metal-responsive transcription factor MTF-1 , nuclear factor kappa B and c-Jun/AP-1 . Increases in caspase-1 and cytokines such as tumor necrosis factor-alpha ( P01375 ) and macrophage inflammatory protein-2 were also evident . In summary , this study profiled the gene expression pattern in mice treated with inorganic arsenicals , which adds to our understanding of acute arsenic poisoning and toxicity . Peroxisome proliferator-activated receptors ( Q07869 ) agonists affect cell viability , apoptosis and expression of cell cycle related proteins in cell lines of glial brain tumors . The nuclear receptors PPARs ( peroxisome proliferator-activated receptors ) are transcription factors activated by specific ligands . PPARs play an important role in carcinogenesis , inflammation , atherosclerosis , lipid metabolism and diabetes . There is evidence that activation of PPARs by specific ligands is able to suppress the growth of different types of human cancer by mechanisms including the growth arrest , apoptosis and induction of differentiation , although the detailed signalling pathways have not been completely elucidated to date . The aim of our study was to determine whether synthetic ligands of PPARalpha and PPARgamma could affect the viability , proliferation , differentiation , apoptosis and expression of some cell cycle related proteins in glial tumor cell lines . The study was performed on human glioblastoma cell lines U-87 MG , T98G , A172 and U-118 MG . Cell lines were treated by ligands of PPARalpha ( bezafibrate , gemfibrozil ) and PPARgamma ( ciglitazone ) . MTT , flow cytometry , TUNEL assay and immunoblotting were used for detection of changes in cell viability , proliferation , differentiation and apoptosis . DB01393 , ciglitazone and gemfibrozil inhibited viability of glioblastoma cell lines . The synthetic ligands significantly reduced or induced the expression of cyclins , P46527 , p21Waf1/Cip1 , MDM-2 , Bcl-2 , Bax , PARP , Caspase 3 , androgen receptors , etc. and did not affect the expression of the differentiation marker P14136 . Flow cytometry confirmed arrest of the cell cycle although the detection of apoptosis was controversial . Apart from hypolipidemic and hypoglycaemic effects , Q07869 ligands may also have significant cytostatic effects of potential use in anticancer treatment . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect .

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interacts_with DB00790?

multiple_choice

[ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . DB00790 : possible use in cancer therapy . Since angiogenesis is essential for the growth of any solid tumor , emerging efforts are being made to develop antiangiogenic therapy . To date , however , no antiangiogenic agent has become widely available for the clinical setting . Angiotensin I-converting enzyme ( P12821 ) inhibitors are commonly used as antihypertensive agents and it has recently been suggested that they decrease the risk of cancer . Studies have found that an P12821 inhibitor , perindopril , is a potent inhibitor of experimental tumor development and angiogenesis at a clinically comparable dose . The potent angiogenic factor , vascular endothelial growth factor ( P15692 ) , is significantly suppressed by perindopril and also inhibits P15692 -induced tumor growth . In vitro studies showed that perindopril is not cytotoxic to either tumor cells or endothelial cells . Since perindopril is already in widespread clinical use without serious side effects , it may represent a potential new strategy for anticancer therapy . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Dual effect of angiotensin-converting enzyme inhibition on angiogenesis in type 1 diabetic mice . OBJECTIVE : We analyzed the beneficial therapeutic effect of angiotensin converting enzyme inhibitor ( ACEI ) on both retinal and hind limb neovascularization in diabetic mice . METHODS AND RESULTS : Diabetic mice ( streptozotocin , 40 mg/kg ) were treated with or without ACEI ( DB00790 , 3 mg/kg per day ) or AT1 receptor blocker ( DB00796 , 20 mg/kg ) for 4 months . Hind limb ischemia was then induced by right femoral artery ligature for 1 additional month . In the ischemic leg , angiographic score , capillary density , and foot perfusion were increased by 2.7 , 2.0-fold , and 1.6-fold , respectively , in ACEI-treated diabetic mice compared with untreated diabetic animals ( P < 0.01 ) . ACEI also raised vascular endothelial growth factor ( P15692 ) protein level by 1.4-fold in ischemic diabetic leg . This ACEI pro-angiogenic effect was totally blunted in diabetic bradykinin B2 receptor-deficient animals , suggesting that it was mediated by the bradykinin pathway . In the diabetic retina , angiotensinogen and P12821 mRNA levels were increased by 2.8-fold and 4.1-fold , respectively ( P < 0.01 versus nondiabetic mice ) , highlighting a local activation of renin-angiotensin system . Diabetes also raised P15692 protein level by 1.5-fold ( P < 0.05 versus nondiabetic mice ) . Treatments with ACEI and AT1 receptor blocker hampered diabetes-induced P15692 upregulation and retinal neovascularization . CONCLUSIONS : P12821 inhibition improved neovascularization in the diabetic ischemic leg through activation of bradykinin signaling , whereas it reduced vessel growth in the diabetic retina through inhibition of overacting Ang II pathway . DB00563 gamma-hydroxamate derivatives as potential dual target antitumor drugs . A series of new aminopteroyl-based hydroxamate derivatives were synthesized and tested in vitro in cell culture models as potential dual target drugs . These compounds were designed to target two families of enzymes , matrix metalloproteinases ( MMP ) and a folate enzyme , dihydrofolate reductase ( P00374 ) . These enzymes are the components of two unrelated cellular pathways and they are often over-expressed in metastasizing tumors . In addition to the synthesis and full structural characterization of the hybrid molecules , we describe their inhibitory activities against a series of MMPs ( P08253 , P09237 , P14780 , P50281 ) and P00374 , as well as their antiproliferative activity in three cancer cell lines . The new hydroxamate derivatives of MTX proved to be effective inhibitors of MMPs and P00374 in the micromolar and nanomolar range , respectively . Furthermore , they showed strong antiproliferative activity against A549 cells ( non-small cell lung carcinoma ) , and PPC-1 and Tsu-Pr1 prostate cancer cell lines . Therefore , based on the present results , these bi-functional drugs may be good candidates to target specific tumors in animal models due to potential combined effects on two pathways crucial for tumor development . The use of microcalorimetry and HPLC for the determination of degradation kinetics and thermodynamic parameters of DB00790 Erbumine in aqueous solutions . DB00790 Erbumine ( O15534 ) is one of the newly used angiotensin-converting enzyme inhibitors ( P12821 inhibitors ) and is used for the treatment of patients with hypertension and symptomatic heart failure . It has two main degradation pathways , i.e. the degradation by hydrolysis and the degradation by cyclization . An isothermal heat conduction microcalorimetry ( MC ) and high pressure liquid chromatography ( HPLC ) were used for the characterization of aqueous solutions of O15534 and its stability properties . The rates of heat evolved during degradation of perindopril were measured by MC as a function of temperature and pH and from these data rate constant and change in enthalpy of the reactions were determined . With the HPLC method the concentration of perindopril and its degradation products were measured as a function of time in aqueous solutions of different pH that were stored at different temperatures . We demonstrated that reactions of degradation of perindopril at observed conditions follow the first order kinetics . The Arrhenius equation for each pH was determined . At pH 6.8 only one degradation pathway is present , i.e. the degradation by hydrolysis . Degradation constants for this pathway calculated from MC data are in good agreement with those obtained from HPLC . MC as a non-specific technique was shown to be useful in studies of O15534 when one reaction was present in the sample and also when more chemical and physical processes were simultaneously running . Neonatal undernutrition and amygdaloid nuclear complex development : an experimental study in the rat . This study describes the morphology of neurons from the basolateral ( P00519 ) , central ( P12821 ) , and medial ( AM ) nuclei of the amygdaloid complex in neonatally undernourished ( U ) and control ( C ) Wistar strain rats . The cells were impregnated with the Golgi-Cox technique and studied at the ages of 12 , 20 , and 40 days postnatally . In the U-pups the neurons of the three nuclei displayed a reduced somatic area compared to that of the C-group on Days 12 and 20 . However , at 40 days , this difference diminished due to a reduction in the somatic area of the C-group . The dendritic area also appeared reduced on Days 12 and 20 in the U-group , but on Day 40 it reached control values . The neurons from P00519 and P12821 suffered a significant decrease in the number of dendritic branches due to undernutrition , but the AM nucleus did not show this change . The data suggest different vulnerability of these amygdaloid nuclei to neonatal undernutrition . The findings also suggest that the abnormal emotional response characteristic of perinatal undernourished rats could have a morphological cause . Inhibition of the renin-angiotensin system improves physiological outcomes in mice with mild or severe cancer cachexia . Cancer cachexia describes the progressive skeletal muscle wasting and weakness associated with many cancers . Cachexia reduces mobility and quality of life and accounts for 20-30 % of all cancer-related deaths . Activation of the renin-angiotensin system causes skeletal muscle wasting and weakness . We tested the hypothesis that treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , perindopril , would enhance whole body and skeletal muscle function in cachectic mice bearing Colon-26 ( C-26 ) tumors . CD2F1 mice received a subcutaneous injection of phosphate buffered saline or C-26 tumor cells inducing either a mild or severe cachexia . The following day , one cohort of C-26 mice began receiving perindopril in their drinking water ( 4 mg kg(-1) day(-1) ) for 21 days . In mild and severe cachexia , perindopril increased measures of whole body function ( grip strength and rotarod ) and reduced fatigue in isolated contracting diaphragm muscle strips ( p < 0.05 ) . In severely cachectic mice , perindopril reduced tumor growth , improved locomotor activity and reduced fatigue of tibialis anterior muscles in situ ( p < 0.05 ) , which was associated with increased oxidative enzyme capacity ( succinate deyhydrogenase , p < 0.05 ) . DB00790 attenuated the increase in Q969Q1 and P05231 mRNA expression and enhanced Akt phosphorylation in severely cachectic mice but neither body nor muscle mass was increased . These findings support the therapeutic potential of P12821 inhibition for enhancing whole body function and reducing fatigue of respiratory muscles in early and late stage cancer cachexia and should be confirmed in future clinical trials . Since P12821 inhibition alone did not enhance body or muscle mass , co-treatment with an anabolic agent may be required to address these aspects of cancer cachexia . Recombinant P17936 inhibits allergic lung inflammation , P15692 production , and vascular leak in a mouse model of asthma . BACKGROUND : Vascular endothelial growth factor ( P15692 ) plays a pro-inflammatory mediator as well as a vascular permeability factor in bronchial asthma . P01308 -like growth factor ( IGF ) -I is also involved in the inflammatory process associated with bronchial asthma and stimulates P15692 expression . The IGF-binding proteins ( IGFBPs ) , especially P17936 , display distinctive properties and can interfere with various biological processes . METHODS : In this study , an ovalbumin ( OVA ) -induced murine model of allergic airway disease was used to investigate which mechanism is implicated in the preventive and therapeutic actions of P17936 administered exogenously on allergen-induced bronchial inflammation and airway hyper-responsiveness , in particular focusing on the regulation of P15692 expression . RESULTS : Administration of recombinant human P17936 to OVA-inhaled mice substantially attenuated the increases in hypoxia-inducible factor ( HIF ) -α activity , P05019 production , and P15692 protein levels in the lung . In addition , the blockade of P05019 action decreased the OVA-induced P15692 expression , airway inflammation , and bronchial hyper-responsiveness . The administration of recombinant human P17936 or CBO-P11 also reduced significantly increases in inflammatory cells , airway hyper-responsiveness , levels of P05112 , P05113 , P35225 , and vascular permeability in the lung of OVA-inhaled mice . Moreover , when recombinant human P17936 was administered after the completion of OVA inhalation , these therapeutic effects of P17936 were also observed . CONCLUSIONS : These results indicate that P17936 administered exogenously may attenuate antigen-induced airway inflammation and hyper-responsiveness through the modulation of vascular leakage and P15692 expression mediated by HIF-1α/HIF-2α signaling as well as P05019 action in allergic airway disease of mice . Update on the biology and therapy of gastrointestinal stromal tumors . BACKGROUND : Gastrointestinal stromal tumors ( GISTs ) , the most common mesenchymal tumors of the gastrointestinal tract , are an example of a disease with an effective , molecularly targeted therapy . METHODS : Published articles and author experience were used to comprehensively define the clinical features , biology , and state-of-the-art therapy of GISTs . RESULTS : GISTs are thought to originate from the neoplastic transformation of the interstitial cells of Cajal , the intestinal pacemaker cells . GISTs commonly have mutations in the kit gene , resulting in a gain-of-function mutation and ligand-independent constitutive activation of the P10721 receptor tyrosine kinase . Successful tyrosine kinase inhibitors target the aberrant pathways that are critical for tumor cell viability . The development of imatinib mesylate ( formerly DB00619 ) in the treatment of metastatic GISTs represents a therapeutic breakthrough . CONCLUSIONS : Progress in the clinical diagnosis has led to an increased recognition of this disease as a distinct clinical entity . Treatment of metastatic GIST with imatinib has led to unprecedented improvements in progression-free and overall survival . The use of imatinib in the preoperative and postoperative treatment of GISTs is an area of intense investigation . Stimulated secretion of endothelial P04275 is accompanied by rapid redistribution to the cell surface of the intracellular granule membrane protein P16109 . We have examined the cell activation-dependent redistribution of the intracellular granule membrane protein P16109 of human endothelial cells . By dual-label immunofluorescence , the distribution of P16109 within cultured human umbilical vein endothelial cells was found to coincide with the distribution of P04275 ( P04275 ) , suggesting that P16109 is located in the membranes of P04275 -containing storage granules . Stimulation of P04275 secretion resulted in an increase in P16109 on the cell surface , as detected by increased binding of the monoclonal antibody P28222 which recognizes the extracytoplasmic domain of P16109 . For each agonist tested ( histamine , thrombin , phorbol 12-myristate 13-acetate , and the calcium ionophore A23187 ) a dose-dependent redistribution of P16109 to the endothelial surface was observed which closely paralleled the dose-dependent secretion of P04275 into the cell supernatant . When cells were maximally stimulated by histamine in the presence of antibody P28222 , a 4-fold increase in P28222 uptake by the cells was observed . This increase occurred rapidly and reached a plateau by 10 min . In contrast , when histamine-stimulated cells were first fixed with paraformaldehyde or chilled to 4 degrees C before addition of antibody P28222 , only a transient increase in cell surface P16109 was detected . Under these conditions of arrested membrane turnover during antibody binding , cell surface P16109 was maximal 3 min after histamine stimulation and then declined to control levels by 20 min . These data suggest that stimulated secretion of P04275 from endothelial cells entails fusion of P04275 -containing storage granules with the plasma membrane . Once inserted into the plasma membrane , P16109 is subsequently removed from the endothelial surface , most likely by an endocytic mechanism . Red wine polyphenols prevent acceleration of neovascularization by angiotensin II in the ischemic rat hindlimb . Studies in both animals and humans indicate that angiogenesis is implicated in the development of atherosclerotic lesions . Thus , inhibition of angiogenesis may provide a novel therapeutic approach for the treatment of atherosclerosis . Because epidemiological studies have indicated an inverse relation between red wine intake and coronary disease , we determined the antiangiogenic potential of red wine polyphenols ( RWPs ) in the ischemic hindlimb model . Neovascularization was accelerated by the chronic infusion of angiotensin II ( Ang II ; 0.1 mg/kg/day ) . RWPs ( 25 mg/kg/day ) or vehicle were administrated in the drinking water 7 days before the ligation . After 21 days , Ang II potentiated the ischemia-induced neovascularization in the hindlimb , as assessed by microangiography and measurement of microvessel density . This effect was associated with an increased formation of reactive oxygen species ( ROS ) and increased expression of hypoxia-inducible factor ( HIF ) -2alpha , endothelial nitric-oxide synthase ( P29474 ) , and vascular endothelial growth factor ( P15692 ) . RWPs intake significantly prevented the angiogenic process , the formation of ROS and nitrated proteins , and the expression HIF-2alpha , P29474 , and P15692 induced by Ang II . Similar preventive effects were observed with the antioxidant and NADPH oxidase inhibitor apocynin . These findings indicate that RWPs have potent antiangiogenic properties in vivo by preventing the expression of proangiogenic factors , including P15692 and P29474 most likely by inhibiting oxidative stress . Thus , the antiangiogenic properties of red wine polyphenols might contribute to their protective effect against coronary disease . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . Chronic methamphetamine exposure alters immune function in normal and retrovirus-infected mice . Methamphetamine ( MA ) abuse represents a growing problem in the USA with an increase of sudden death . To evaluate the immune function alterations due to chronic methamphetamine use , we examined C57BL/C mice with LP-BM5 retrovirus infection plus methamphetamine exposure . Mice were randomly assigned to the following groups : placebo , placebo retrovirus-infected , uninfected MA treated and retrovirus-infected MA treated . Placebo , MA-treated groups were intraperitoneally injected with saline , MA , respectively , with a gradually increasing dose from 15 to 40 mg/kg for 12 weeks ( 5 days/week ) . Con A- and LPS-induced mitogenesis of splenocytes , cytokine production by splenocytes culture and lipid peroxides in the liver were measured . Heart tissue histopathology was analyzed in all the groups with murine cytomegalovirus ( CMV ) superinfection . Our data showed that MA treatment significantly decreased production of P60568 and interferon gamma ( P01579 ) in uninfected mice but did not further suppress the reduced Th1 cytokines in retrovirus-infected mice . There were no significant effects on cytokines P05112 and P05231 . However , tumor necrosis factor ( P01375 ) was significantly increased in both uninfected and infected mice due to MA treatment . Lipid peroxides in liver were significantly increased both in uninfected and retrovirus-infected mice due to MA exposure . DB00163 levels in liver were significantly decreased in uninfected mice due to MA treatment . CMV superinfection greatly increased the cardiac lesions in retrovirus-infected mice while no significant histopathology changes were detected due to MA treatment . Our data suggest that MA has immunomodulation activity , suppressing Th1 cytokine production and enhancing some Th2 cytokine secretion , as well as increasing lipid peroxides in uninfected mice . The interaction between LP-BM5 and MA remains unclear . DB00790 decreases P wave dispersion in patients with stage 1 hypertension . INTRODUCTION : P12821 inhibitors prevent atrial fibrillation episodes by effective control of blood pressure and improving electrical and structural remodelling in the atria . Increased P wave dispersion ( P35670 ) is a non-invasive electrocardiographic marker for paroxysmal atrial fibrillation . The aim of the study was to evaluate the effect of perindopril treatment on P35670 in hypertensive patients . METHODS : Forty-eight hypertensive patients ( mean age 57.4+/-11.8 years , 18 men ) were included . Blood pressure values were determined and 12-lead electrocardiograms were recorded at the beginning and at the first week , first month , third month and sixth month of the perindopril treatment.The difference between maximum and minimum P wave durations was calculated as P35670 . RESULTS : PWDs were significantly shortened at the first , third and sixth months ( 41.7+/-8.8 ms , Q04695 +/-6.9 ms and 38.3+/-7.1 ms , respectively ) compared with baseline and first-week measurements ( 54.3+/-9.2 ms and 49.0+/-9.1 ms , respectively , p < 0.001 ) . Baseline P35670 was correlated with body mass index ( r=0.32 , p=0.026 ) , while P35670 at the sixth month of treatment was significantly correlated with left atrial volume index ( r=0.30 , p=0.042 ) . Multiple linear regression analysis revealed that P35670 at the sixth month was related to baseline P35670 ( p=0.001 ) . CONCLUSION : DB00790 treatment significantly reduced P35670 in hypertensive patients . Nonlinkage of bipolar illness to tyrosine hydroxylase , tyrosinase , and D2 and D4 dopamine receptor genes on chromosome 11 . OBJECTIVE : Previous linkage and allelic association studies using DNA polymorphisms , cosegregation of cytogenetic abnormalities with psychiatric illness , and assignment of genes involved in neutotransmitter metabolism suggested that chromosome 11 may harbor a gene predisposing to bipolar illness . The authors examined linkage in the families of 14 probands with bipolar illness , with the candidate genes tyrosine hydroxylase ( TH ) , D4 dopamine receptor ( P21917 ) at 11p15 , tyrosinase ( P14679 ) at 11q14-q21 , and D2 dopamine receptor ( P14416 ) at 11q22-q23 , as well as with the c-Harvey-ras oncogene ( P01112 ) and insulin gene ( P01308 ) , both located at 11p15 , a region that previously showed linkage to bipolar illness . METHOD : The genetic data were analyzed with both lod score analysis ( parametric ) and affected-sib-pair analysis ( nonparametric ) ; both narrow and broad definitions of the clinical phenotype were used . Further influences of diagnostic uncertainties were accounted for by using diagnostic probability classes weighing the stability of each phenotype . RESULTS : Two-point linkage results excluded close linkage of bipolar illness to each candidate gene ; negative results were also obtained when the narrow definition of the clinical phenotype was used . Moreover , multipoint linkage analysis of P01112 and P01308 excluded the 11p15 region encompassing both P21917 and TH . In agreement with the negative linkage results , affected-sib-pair analysis did not show preferential sharing of marker alleles at any of the candidate genes . CONCLUSIONS : The negative results obtained under different genetic models exclude a frequent role for P21917 , TH , P14679 , and P14416 in the pathogenesis of bipolar illness . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Effects of PEG-coupled interleukin-2 on rat lung lavage parameters . The recombinant lymphokine interleukin-2 ( P60568 ) has activity in renal cell carcinoma , melanoma , and other cancers . A side effect of P60568 use is a " capillary leak phenomenon " which is purported to be related to endothelial effects of P60568 itself or to cells activated by P60568 . We studied P60568 effects on rat lung lavage parameters to determine whether endothelial damage occurred . The specific endpoints were 125I-albumin extravasation , lavage protein , and lavage angiotensin-converting enzyme ( P12821 ) activity . To ensure sensitivity of these endpoints , we used the known endothelial toxicant thiourea , which increases lung lavage protein and lavage P12821 . We found that both PEG P60568 and thiourea increased the amount of protein and 125-I flux into the lavage . However , although thiourea increased lavage P12821 , PEG P60568 did not . These results suggest that PEG P60568 can increase protein and iodine flux across the endothelium without causing cell injury . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . [ Association and interaction of AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 and P35354 genes on the risk of hypertension in Antioquian population ] . INTRODUCTION : Hypertension is a multifactorial disease influenced by genetic and environmental components , with its prevalence varying across ethnic groups . Manifold studies on blood pressure regulatory system genes have been carried out -such as the renin-angiotensin-aldosterone system , the sympathetic nervous system , endothelial factor , and sodium balance- , but the results yielded were inconsistent among populations . OBJECTIVES : To evaluate the effect of both variants in genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 P35354 , and the result of the individual ancestry component on hypertension and blood pressure levels among population in Antioquia . METHODS AND MATERIALS : 107 cases and 253 controls were genotyped for 12 variants on genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 y P35354 , and for 20 ancestry informative markers . The association of polymorphisms and their interactions , and the association of ancestral genetic composition with hypertension and blood pressure levels were examined . RESULTS : Genes P35612 , rs4852706 ( OR=3.0 ; p=0.023 ) ; P21728 , rs686 ( OR=0.38 ; p=0.012 ) and P07550 , rs1042718 ( OR=10.0 ; p=0.008 ) ; as well as genotypic combinations of P21728 and P30556 ; AGT and P35611 ; and P35611 to P20020 and P35354 were associated to hypertension . The Amerindian ancestry component was associated to some decrease in diastolic blood pressure . CONCLUSION : Variants on genes P35612 , P21728 , P07550 , P30556 , AGT , P35611 , P20020 and P35354 individually or interacting , are associated to hypertension . The Amerindian ancestry component has an effect on blood pressure . P37840 activates microglia by inducing the expressions of matrix metalloproteinases and the subsequent activation of protease-activated receptor-1 . The mutation or overexpression of alpha-synuclein protein plays a pivotal role in the pathogenesis of Parkinson 's disease . In our preliminary experiments , we found that alpha-synuclein induced the expression of matrix metalloproteinases ( MMPs ) ( P03956 , -3 , -8 , and -9 ) in rat primary cultured microglia . Thus , the current study was undertaken to determine the roles of MMPs in alpha-synuclein-induced microglial activation . The inhibition of P08254 , -8 , or -9 significantly reduced NO and reactive oxygen species levels and suppressed the expression of P01375 and IL-1beta . Notably , P22894 inhibitor suppressed P01375 production more efficaciously than P08254 or P14780 inhibitors . Inhibition of P08254 or -9 also suppressed the activities of MAPK , NF-kappaB , and AP-1 . Previously , protease-activated receptor-1 ( P25116 ) has been associated with the actions of MMPs , and thus , we further investigated the role of P25116 in alpha-synuclein-induced inflammatory reactions . A P25116 -specific inhibitor and a P25116 antagonist significantly suppressed cytokine levels , and NO and reactive oxygen species production in alpha-synuclein-treated microglia . Subsequent P25116 cleavage assay revealed that P08254 , -8 , and -9 , but not alpha-synuclein , cleaved the synthetic peptide containing conventional P25116 cleavage sites . These results suggest that MMPs secreted by alpha-synuclein-stimulated microglia activate P25116 and amplify microglial inflammatory signals in an autocrine or paracrine manner . Furthermore , our findings suggest that modulation of the activities of MMPs and/or P25116 may provide a new therapeutic strategy for Parkinson 's disease . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . P12821 activity is involved in the mechanism of increased endogenous nitric oxide synthase inhibitor in patients with type 2 diabetes mellitus . The renin-angiotensin system plays an important role in the elevation of asymmetric dimethylarginine ( DB01686 ) , an endogenous inhibitor of nitric oxide synthase , in hypertensive patients , so the present study was designed to examine whether angiotensin-converting enzyme ( P12821 ) activity is also involved in the mechanism of DB01686 elevation in type 2 diabetes mellitus ( NIDDM ) . A crossover study was performed to determine if P12821 inhibition with perindopril ( 4 mg/day ) for 4 weeks decreases serum DB01686 concentration and plasma P04275 ( P04275 ) level ( a marker of endothelial injury ) in 11 patients with NIDDM . None of the patients was treated with insulin or oral hypoglycemic drugs , and none had major diabetic complications . Before the protocol began , serum DB01686 and plasma P04275 were significantly higher in the 11 NIDDM patients , when compared with 8 control subjects without diabetes . DB00790 did not affect blood pressure or glucose metabolism , but did significantly decrease serum DB01686 and plasma P04275 . These results suggest that endothelial injury associated with DB01686 elevation may be present even in patients with non-complicated NIDDM , and that increased activity of P12821 may be involved in such endothelial dysfunction . Vascular endothelial growth factor expression and glomerular endothelial cell loss in the remnant kidney model . BACKGROUND : Vascular endothelial growth factor ( P15692 ) is constitutively expressed in the glomerulus where it may have a role in the maintenance of capillary endothelial cell integrity . The present study sought to examine changes in P15692 expression in a model of progressive renal disease and to assess the effects of angiotensin converting enzyme ( P12821 ) inhibition . METHODS : Subtotal nephrectomized ( STNx ) rats were randomly assigned to receive vehicle ( n=10 ) or the P12821 inhibitor perindopril ( 8 mg/l drinking water ) for 12 weeks duration ( n=10 ) . Sham-operated rats were used as controls ( n=10 ) . Glomerular capillary endothelial cell density was evaluated by immunostaining for the pan-endothelial cell marker Q06609 -1 and P15692 expression was assessed by quantitative in situ hybridization . RESULTS : In STNx rats glomerular capillary endothelial cell density was reduced to 19 % that of sham rats ( P < 0.01 ) with a concomitant reduction in glomerular P15692 expression , also to 19 % of sham rats ( P < 0.01 ) . DB00790 treatment was associated with normalization of both capillary endothelial cell density and glomerular P15692 mRNA . CONCLUSIONS : Reduction in glomerular P15692 expression is a feature of the renal pathology that follows subtotal nephrectomy . In the context of the known functions of this growth factor , these findings suggest that diminution in P15692 may contribute to the demonstrated loss of glomerular endothelium that develops in this model of progressive renal disease . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . [ An effect of perindopril on the level of tumor necrosis factor-alpha and matrix metalloproteinase-9 in peripheral blood in the acute period of atherothrombotic ischemic stroke and myocardial infarction ] . Twenty-nine patients with acute atherothrombotic ischemic stroke and 36 patients with acute Q-wave myocardial infarction have been studied . Each group has been stratified into 2 subgroups : patients of subgroups A received an P12821 inhibitor perindopril in the complex therapy from the 1st day of disease . Patients of subgroups B were not assigned to this drug . Along with routine tests , the level of tumor necrosis factor-alpha and matrix metalloproteinase-9 ( P14780 ) measured with ELISA using test-systems ( Q02223 Diagnostics , USA ) and reagents ( R & D , England ) have been determined . The administration of perindopril did not cause side-effects , including arterial hypotonia after the first dosage , in patients in the acute period of atherothrombotic ischemic stroke and myocardial infarction . DB00790 may decrease the activity of P14780 in these patients and produces an anticytokine effect . Some similar mechanisms of ischemic lesions of the heart and the brain and a commonness of biochemical " response " to the same medical intervention ( the administration of an P12821 inhibitor perindopril ) in patients of both groups were found . The results support the pathogenetic validity of perindopril therapy in the secondary prevention of ischemic stroke and myocardial infarction . Association of angiotensin-converting enzyme gene I/D polymorphism with steroid responsiveness in childhood nephrotic syndrome . The aim of the study was to study the distribution of angiotensin-converting enzyme ( P12821 ) gene insertion/deletion ( I/D ) polymorphism , and its association with steroid responsiveness in children with idiopathic nephrotic syndrome ( P01308 ) . One hundred twenty-five children with P01308 were classified into two groups : steroid-sensitive nephrotic syndrome ( SSNS : n = 90 ) and steroid-resistant nephrotic syndrome ( SRNS : n=35 ) . The control group consisted of 150 unrelated healthy children . Genomic DNA was extracted from peripheral leucocytes by the standard salting-out method . P12821 genotyping was performed and P12821 genotypes DD , ID , and II were compared between different groups . The frequency distribution of the DD genotype was significantly increased in children with P01308 compared to control subjects ( P = 0.0012 ) while the difference was not significant ( P = 0.071 ) between SSNS and control subjects . The frequency distribution of the DD genotype was significantly high in the SRNS group compared to control subjects ( P < 0.0001 ) . The distribution of the DD genotype was high in SRNS compared to SSNS group patients ( P = 0.016 ) . In conclusion , the presence of the DD genotype may predict risk for steroid resistance in childhood P01308 . In utero exposure to maternal diets containing soy protein isolate , but not genistein alone , protects young adult rat offspring from P48645 -induced mammary tumorigenesis . The linkage of nutrition and cancer prevention is an intriguing concept that is gaining widespread support . Here , we investigated the influence of developmental context on dietary protection against tumorigenesis initiated by the direct-acting carcinogen N-methyl-N-nitrosourea ( P48645 ) , and examined potential mechanisms underlying these effects . Rats were exposed only in utero or for lifetime to American Institute of Nutrition-93G diets made with casein ( CAS ) , soy protein isolate ( SPI ) or CAS supplemented with genistein ( GEN ) . Mammary glands of post-natal day ( P01160 ) 50 rats prior to P48645 administration were examined for apoptotic status , pro-apoptotic gene expression and immunoreactive phosphatase and tensin homolog deleted on chromosome ten ( P60484 ) and epithelial cadherin ( P12830 ) levels , whereas mammary tumor parameters were evaluated 99 days post- P48645 . Animals exposed only in utero to SPI had increased tumor latency , decreased tumor multiplicity and lower higher grade tumors , than those fed CAS . In utero exposure to GEN resulted in similar tumor parameters as the CAS group , whereas lifetime SPI exposure decreased tumor incidence that was not mimicked by in utero exposure alone . Mammary glands of PND50 rats fed lifetime SPI had increased terminal end bud apoptotic status and P60484 expression , than the other diet groups . Rats exposed only in utero to SPI or GEN had higher membrane P12830 in mammary structures than those lifetime-fed CAS or SPI . Thus , limited exposure during gestation to SPI can positively influence resistance to chemically induced mammary tumorigenesis later in life . Preventative strategies against mammary and other types of cancer might be uncovered by refinement of the developmental window for dietary factor exposure . Effects of cytokines on P15692 expression and secretion by human first trimester trophoblast cell line . PROBLEM : The mechanism through which vascular endothelial growth factor ( P15692 ) regulation occurs at the feto-maternal interface is poorly understood . The aim of this study was to investigate the effects of various cytokines on P15692 expression and secretion by trophoblast cells . METHOD OF STUDY : We investigated the effects of cytokines on P15692 expression in human first trimester trophoblast cell line by analyzing P15692 messenger RNA ( mRNA ) by reverse transcription-polymerase chain reaction and P15692 protein secretion by enzyme linked immunosorbent assay . RESULTS : The trophoblast cells expressed P15692 mRNA constitutively and the main subtypes were identified as VEGF121 and VEGF165 . When cultured in the presence of interferon ( IFN ) -gamma , interleukin ( IL ) - 1beta , tumor necrosis factor ( P01375 ) -alpha , P60568 , or P22301 , P15692 mRNA expression was found to be significantly increased by IL-1beta , P01579 and P01375 but to be unaffected by P60568 and P22301 . Moreover , P15692 secretion was most significantly increased by P01579 treatment . CONCLUSION : These results suggest that IL-1beta , P01579 , and P01375 may regulate the production of P15692 in early gestational trophoblasts . Pharmacophore-based virtual screening versus docking-based virtual screening : a benchmark comparison against eight targets . AIM : This study was conducted to compare the efficiencies of two virtual screening approaches , pharmacophore-based virtual screening ( PBVS ) and docking-based virtual screening ( DBVS ) methods . METHODS : All virtual screens were performed on two data sets of small molecules with both actives and decoys against eight structurally diverse protein targets , namely angiotensin converting enzyme ( P12821 ) , acetylcholinesterase ( P22303 ) , androgen receptor ( AR ) , D-alanyl-D-alanine carboxypeptidase ( DacA ) , dihydrofolate reductase ( P00374 ) , estrogen receptors alpha ( ERalpha ) , HIV-1 protease ( HIV-pr ) , and thymidine kinase ( TK ) . Each pharmacophore model was constructed based on several X-ray structures of protein-ligand complexes . Virtual screens were performed using four screening standards , the program Catalyst for PBVS and three docking programs ( DOCK , GOLD and Glide ) for DBVS . RESULTS : Of the sixteen sets of virtual screens ( one target versus two testing databases ) , the enrichment factors of fourteen cases using the PBVS method were higher than those using DBVS methods . The average hit rates over the eight targets at 2 % and 5 % of the highest ranks of the entire databases for PBVS are much higher than those for DBVS . CONCLUSION : The PBVS method outperformed DBVS methods in retrieving actives from the databases in our tested targets , and is a powerful method in drug discovery . P37840 A30P point-mutation generates age-dependent nigrostriatal deficiency in mice . Lewy bodies are mainly composed of alpha-synuclein ( P37840 ) and specific mutations in P37840 gene are related to familial forms of Parkinson 's disease ( PD ) . The purpose of our study was to generate a mouse line with A30P knock-in point mutation in P37840 gene and to test if a single point-mutation is able to turn otherwise normal P37840 into a toxic form . The behavioral profile of P37840 A30P mice was followed for 16 months . Generally , these mice are healthy and viable without any obvious abnormalities . Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function ( ink-test and beam walk ) . In other tests ( motility boxes , rotarod ) mice continuously performed normally . Moreover , P37840 A30P mice expressed the altered sensitivity to Q05940 inhibitor reserpine , possibly reflecting a functional deficiency of dopamine . Indeed , mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum , and reduced levels of dopamine in the mesolimbic system . The present study confirms that P37840 plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance . The generated mouse line has a potential to become a model for PD with comparable time course and phenotype . Bacterial translocation in cirrhotic rats stimulates P29474 -derived NO production and impairs mesenteric vascular contractility . DB00435 ( NO ) has been implicated in the arterial vasodilation and associated vascular hyporesponsiveness to vasoconstrictors observed in liver cirrhosis . Bacteria , potent activators of NO and P01375 synthesis , are found in the mesenteric lymph nodes ( MLNs ) of ascitic cirrhotic rats . Here , we investigated the impact of bacterial translocation ( BT ) to MLNs on P01375 production , vascular NO release , and contractility in the mesenteric vasculature of ascitic cirrhotic rats . Vascular response to the alpha-adrenoagonist methoxamine , which is diminished in the superior mesenteric arterial beds of cirrhotic rats , is further blunted in the presence of BT . BT promoted vascular NO release in cirrhotic rats , an effect that depended on pressure-induced shear stress and was blocked by the NO inhibitor N(omega)-nitro-L-arginine . Removing the endothelium had the same effect . Endothelial NO synthase ( P29474 ) , but not the inducible isoform ( P35228 ) , was present in mesenteric vasculature of cirrhotic rats with and without BT , and its expression was enhanced compared with controls . P01375 was induced in MLNs by BT and accumulated in parallel in the serum . This P01375 production was associated with elevated levels of tetrahydrobiopterin ( BH(4) ) , a P01375 -stimulated cofactor and enhancer of P29474 -derived NO biosynthesis and NOS activity in mesenteric vasculature . These findings establish a link between BT to MLNs and increased P01375 production and elevated BH(4) levels enhancing P29474 -derived NO overproduction , further impairing contractility in the cirrhotic mesenteric vasculature . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Cellular mechanisms of the hemostatic effects of desmopressin ( DB00035 ) . The synthetic analog of vasopressin desmopressin ( DB00035 ) is widely used for the treatment of patients with von Willebrand disease ( VWD ) , hemophilia A , several platelet disorders , and uremic bleeding . DB00035 induces an increase in plasma levels of P04275 ( P04275 ) , coagulation factor VIII ( FVIII ) , and tissue plasminogen activator ( t-PA ) . It also has a vasodilatory action . In spite of its extensive clinical use , its cellular mechanism of action remains incompletely understood . Its effect on P04275 and t-PA as well as its vasodilatory effect are likely explained by a direct action on the endothelium , via activation of endothelial vasopressin P30518 receptor and DB02527 -mediated signaling . This leads to exocytosis from Weibel Palade bodies where both P04275 and t-PA are stored , as well as to nitric oxide ( NO ) production via activation of endothelial NO synthase . The mechanism of action of DB00035 on FVIII plasma levels remains to be elucidated . The hemostatic effect of DB00035 likely involves additional cellular effects that remain to be discovered . Role of angiotensin II in the remodeling induced by a chronic increase in flow in rat mesenteric resistance arteries . Angiotensin II is a potent growth factor involved in arterial wall homeostasis . In resistance arteries , chronic increases in blood flow induce a rise in diameter associated with arterial wall hypertrophy . Nevertheless , the role of angiotensin II in this remodeling is unknown . We investigated the effect of blocking angiotensin II production or receptor activation on flow-induced remodeling of mesenteric resistance arteries . Arteries were ligated in vivo to generate high-flow arteries compared with normal flow ( control ) vessels located at a distance . Arteries were isolated after 1 week for in vitro analysis . Arterial diameter , media surface , endothelial NO synthase expression , superoxide production , and extracellular signal-regulated kinase 1/2 phosphorylation were higher in high-flow than in control arteries . P12821 inhibition ( perindopril ) and angiotensin II type 1 receptor blockade ( candesartan ) prevented arterial wall hypertrophy without affecting diameter enlargement . The nonselective vasodilator hydralazine had no effect on remodeling . Although perindopril and candesartan increased endothelial NO synthase expression in high-flow arteries , hypertrophy remained in rats treated with N(G)-nitro-l-arginine methyl ester and mice lacking endothelial NO synthase . DB00790 and candesartan reduced oxidative stress in high-flow arteries , but superoxide scavenging did not prevent hypertrophy . Both Tempol and the absence of endothelial NO synthase prevented the rise in diameter in high-flow vessels . P27361 /2 activation in high-flow arteries was prevented by perindopril and candesartan and not by hydralazine . P27361 /2 inhibition in vivo ( U0126 ) prevented hypertrophy in high-flow arteries . Thus , a chronic rise in blood flow in resistance arteries induces a diameter enlargement involving NO and superoxide , whereas hypertrophy was associated with extracellular signal-regulated kinase 1/2 activation by angiotensin II . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . The use of PEGT/ P10721 as a dermal scaffold for skin tissue engineering . Human skin equivalents ( HSEs ) were engineered using biodegradable-segmented copolymer PEGT/ P10721 as a dermal scaffold . As control groups , fibroblast-populated de-epidermized dermis , collagen , fibrin and hybrid PEGT/ P10721 -collagen matrices were used . Two different approaches were used to generate full-thickness HSE . In the 1-step approach , keratinocytes were seeded onto the fibroblast-populated scaffolds and cultured at the air-liquid ( A/L ) interface . In the 2-step approach , fully differentiated epidermal sheets were transferred onto fibroblast-populated scaffolds and cultured at the A/L . In a 1-step procedure , keratinocytes migrated into the porous PEGT/ P10721 scaffold . This was prevented by incorporating fibroblast-populated collagen into the pores of the PEGT/ P10721 matrix or using the 2-step procedure . Under all experimental conditions , fully differentiated stratified epidermis and basement membrane was formed . Differences in K6 , P08779 , Q04695 , collagen type VII , laminin 5 and nidogen staining were observed . In HSE generated with PEGT/ P10721 , the expression of these keratins was higher , and the deposition of collagen type VII , laminin 5 and nidogen at the epidermal/matrix junction was retarded compared to control HSEs . Our results illustrate that the copolymer PEGT/ P10721 is a suitable scaffold for the 2-step procedure , whereas the incorporation of fibroblast-populated collagen or fibrin into the pores of the scaffold is required for the 1-step procedure . P00797 -angiotensin system modulation : the weight of evidence . Modulation of the renin-angiotensin system is considered to be the most complete way to manage high-risk patients including those with hypertension . P12821 ( P12821 ) inhibitors are effective at reducing the morbidity and mortality of patients with overt clinical heart failure , asymptomatic left ventricular dysfunction , and uncomplicated myocardial infarction . Furthermore , recent trials like the Heart Outcomes Prevention Evaluations ( HOPE ) study and the EUropean trial on Reduction Of cardiac events with DB00790 in stable coronary Artery disease ( EUROPA ) support extending the use of P12821 inhibitors to the routine/first-line treatment of patients with an increased global cardiovascular risk . Although some investigators have seen the development of angiotensin II receptor blockers ( ARBs ) as a more effective and tolerable way of reproducing the benefits of P12821 inhibition , there remain important concerns regarding the distinct pharmacologic profiles and modes of action of these two classes of drugs . Careful evaluation of data from recent large-scale studies revealed that , unlike P12821 inhibitors , ARBs are either neutral or may actually increase rates of myocardial infarction despite similar levels of blood pressure reduction . The fact that this effect is most apparent when ARBs are compared with placebo in the absence of concomitant P12821 inhibitors suggests that differential effects on the angiotensin II type 2 ( AT(2) ) receptors may be important . Other important pharmacologic differences are also known to be present and may be of direct relevance . The weight of available evidence therefore supports the use of appropriate P12821 inhibitor regimens , although not ARBs , in the treatment of global cardiovascular risk . Evaluation of antileukaemic effects of rapamycin in patients with imatinib-resistant chronic myeloid leukaemia . BACKGROUND : Recent data suggest that the mammalian target of rapamycin ( P42345 ) is involved in the regulation of growth of neoplastic cells in chronic myeloid leukaemia ( CML ) . PATIENTS AND METHODS : We treated six patients with imatinib-resistant CML in haematological relapse ( leukocytes > 20,000 microL(-1) ) with rapamycin at 2 mg per os daily for 14 consecutive days , with dose-adjustment allowed to reach a target rapamycin serum concentration of 10-20 pg mL(-1) . RESULTS : A major leukocyte response with decrease to less than 10,000 microL(-1) was obtained in two patients , and a minor transient response was seen in two other patients . In responding patients , we also observed a decrease in vascular endothelial growth factor ( P15692 ) mRNA levels in circulating leukaemic cells . Side effects during rapamycin treatment were mild in most patients . In one patient , pneumonia developed . DB00877 was also found to counteract growth of CML cells in vitro as determined by (3)H-thymidine incorporation . Moreover , rapamycin inhibited the growth of Ba/ P13726 cells exhibiting various imatinib-resistant mutants of P11274 / P00519 , including the T315I variant that exhibits resistance against most currently available P11274 / P00519 kinase inhibitors . CONCLUSIONS : DB00877 shows antileukaemic effects in imatinib-resistant CML in vitro and in vivo . Larger trials with rapamycin or rapamycin-derivatives in combination with other targeted drugs are warranted to further determine clinical efficacy in CML . REV-ERBα inhibits the P35354 expression in bovine uterus endometrium stromal and epithelial cells exposed to ovarian steroids . The nuclear receptor REV-ERBα ( encoded by P20393 ) has a critical role in metabolism and physiology as well as circadian rhythm . Here , we investigated the possible contribution of clock genes including P20393 to the secretion of prostaglandin F2α ( PGF2α ) from bovine uterine stromal ( USCs ) and epithelial cells ( UECs ) by modulating the expression of P35354 . The circadian oscillation of clock genes in the cells was weak compared with that reported in rodents , but the expression of O00327 , O15534 , and P20393 was changed temporally by treatment with ovarian steroids . Significant expression of clock genes including P20393 was detected in USCs exposed to progesterone . P20393 was also significantly expressed in UECs exposed to estradiol . The expression of P35354 was suppressed in USCs exposed to progesterone , while the expression was initially suppressed in UECs exposed to estradiol and then increased after long-term exposure to estradiol . O00327 knockdown with specific siRNA caused a significant decrease in the transcript levels of P20393 and P35354 in USCs , but not in UECs . The production of PGF2α also decreased in USCs after O00327 knockdown , while its level did not significantly change in UECs . The transcript level of P35354 was increased by treatment with the antagonist of REV-ERBα in both cell types , but the agonist was ineffective . In these two cell types treated with the agonist or antagonist , the PGF2α production coincided well with the P35354 expression . Collectively , these results indicate that REV-ERBα plays an inhibitory role in the expression of P35354 in both bovine USCs and UECs treated with ovarian steroids . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . Activation of P00533 promotes squamous carcinoma SCC10A cell migration and invasion via inducing EMT-like phenotype change and P14780 -mediated degradation of P12830 . P00533 is a potent stimulator of invasion and metastasis in head and neck squamous cell carcinomas ( HNSCC ) . However , the mechanism by which P00533 may stimulate tumor cell invasion and metastasis still need to be elucidated . In this study , we showed that activation of P00533 by P01133 in HNSCC cell line SCC10A enhanced cell migration and invasion , and induced loss of epitheloid phenotype in parallel with downregulation of P12830 and upregulation of P19022 and vimentin , indicating that P00533 promoted SCC10A cell migration and invasion possibly by an epithelial to mesenchymal transition ( EMT ) -like phenotype change . Interestingly , activation of P00533 by P01133 induced production of matrix metalloproteinase-9 ( P14780 ) and soluble P12830 ( sE-cad ) , and knockdown of P14780 by siRNA inhibited sE-cad production induced by P01133 in SCC10A . Moreover , both P14780 knockdown and P12830 overexpression inhibited cell migration and invasion induced by P01133 in SCC10A . The results indicate that P00533 activation promoted cell migration and invasion through inducing P14780 -mediated degradation of P12830 into sE-cad . Pharmacologic inhibition of P00533 , MEK , and PI3K kinase activity in SCC10A reduced phosphorylated levels of P27361 /2 and AKT , production of P14780 and sE-cad , cell migration and invasion , and expressional changes of EMT markers ( P12830 and P19022 ) induced by P01133 , indicating that P00533 activation promotes cell migration and invasion via P27361 /2 and PI3K-regulated P14780 / P12830 signaling pathways . Taken together , the data suggest that P00533 activation promotes HNSCC SCC10A cell migration and invasion by inducing EMT-like phenotype change and P14780 -mediated degradation of P12830 into sE-cad related to activation of P27361 /2 and PI3K signaling pathways . Different effects of perindopril and enalapril on monocyte cytokine release in coronary artery disease patients with normal blood pressure . BACKGROUND : Favorable effects of angiotensin-converting enzyme ( P12821 ) inhibitor treatment on the incidence of cardiovascular and cerebrovascular mortality and morbidity are not limited to patients with elevated blood pressure . As suggested by our previous results , the physicochemical and pharmacokinetic differences between drugs may markedly contribute to the strength of pleiotropic effects of P12821 inhibitors . METHODS : The present study was aimed at comparing the effects of serum- and tissue-type P12821 inhibitors on monocyte release of proinflammatory cytokines in normotensive patients with stable coronary artery disease . The participants were randomized to 90-day treatment with enalapril ( 20 mg daily , n = 29 ) , perindopril ( 4 mg daily , n = 27 ) or placebo ( n = 28 ) . Plasma levels of lipids , glucose , insulin and high sensitivity P02741 ( hsCRP ) , as well as monocyte release of proinflammatory cytokines were determined before and after 30 days of therapy , and at the end of the treatment . RESULTS : Lipopolysaccharide-stimulated monocytes from normotensive patients with stable coronary artery disease released significantly more P01375 -α , interleukin-1β and monocyte chemoattractant protein-1 in comparison with monocytes from 23 matched control subjects . Their baseline hsCRP levels were also higher . DB00790 reversed the disease-induced changes in cytokine release and reduced plasma hsCRP , while the effect of enalapril was much more limited . The effect on both drugs on cytokine release was stronger in insulin-resistant than insulin-sensitive subjects . CONCLUSIONS : Our results indicate that perindopril is superior to enalapril in producing monocyte-suppressing and systemic anti-inflammatory effects in normotensive patients with coronary artery disease . This action may contribute to the clinical effectiveness of tissue P12821 inhibitors in the therapy of atherosclerosis-related disorders , particularly in insulin-resistant subjects . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . DB00184 induces cell proliferation , invasion and epithelial-mesenchymal transition in a variety of human cancer cell lines . Cigarette smoking is strongly correlated with the onset of nonsmall cell lung cancer ( NSCLC ) . DB00184 , an active component of cigarettes , has been found to induce proliferation of lung cancer cell lines . In addition , nicotine can induce angiogenesis and confer resistance to apoptosis . All these events are mediated through the nicotinic acetylcholine receptors ( nAChRs ) on lung cancer cells . In this study , we demonstrate that nicotine can promote anchorage-independent growth in NSCLCs . In addition , nicotine also induces morphological changes characteristic of a migratory , invasive phenotype in NSCLCs on collagen gel . These morphological changes were similar to those induced by the promigratory growth factor P15692 . The proinvasive effects of nicotine were mediated by alpha7-nAChRs on NSCLCs . RT-PCR analysis showed that the alpha7-nAChRs were also expressed on human breast cancer and pancreatic cancer cell lines . DB00184 was found to promote proliferation and invasion in human breast cancer . The proinvasive effects of nicotine were mediated via a nAChR , Src and calcium-dependent signaling pathway in breast cancer cells . In a similar fashion , nicotine could also induce proliferation and invasion of Aspc1 pancreatic cancer cells . Most importantly , nicotine could induce changes in gene expression consistent with epithelial to mesenchymal transition ( EMT ) , characterized by reduction of epithelial markers like P12830 expression , ZO-1 staining and concomitant increase in levels of mesenchymal proteins like vimentin and fibronectin in human breast and lung cancer cells . Therefore , it is probable that the ability of nicotine to induce invasion and EMT may contribute to the progression of breast and lung cancers . P00797 -angiotensin system is involved in the mechanism of increased serum asymmetric dimethylarginine in essential hypertension . Endothelium-dependent/nitric oxide ( NO ) -mediated vasodilation is impaired in hypertensive individuals . DB01686 ( DB01686 ) , an endogenous inhibitor of NO synthase , is synthesized by many types of cells including vascular endothelial cells . The serum level of DB01686 is elevated in patients with essential hypertension , but the mechanism for this increase is unknown . Therefore , the present study examined whether the renin-angiotensin system ( DB01367 ) is involved . Patients with essential hypertension [ systolic blood pressure ( BP ) > 160 mmHg and/or diastolic BP > 95 mmHg ] were randomized to an angiotensin-converting enzyme ( P12821 ) inhibitor treatment group ( perindopril , 4mg/day for 4 weeks , n = 7 ) , an angiotensin II type 1 ( AT1 ) receptor antagonist treatment group ( losartan , 50 mg/day for 4 weeks , n = 7 ) or a beta-blocker treatment group ( bisoprolol , 5 mg/day for 4 weeks , n = 7 ) . Before and after the treatment , BP , serum concentration of DB01686 and plasma concentration of P04275 ( P04275 , a biological marker of endothelial injury ) were measured . DB00790 , losartan and bisoprolol decreased BP to a similar extent , and either perindopril or losartan , but not bisoprolol , significantly decreased serum DB01686 and plasma P04275 . These findings suggest that the DB01367 may contribute to the mechanism of increased serum DB01686 as well as to the endothelial injury observed in hypertensive patients . The vasculoprotective actions of P12821 inhibitors or AT1 receptor antagonists may be explained at least in part by amelioration of the endothelial injury through a decrease in the serum DB01686 concentration . Cardiac protective effect of Astragalus on viral myocarditis mice : comparison with DB00790 . In clinical practice , Astragali Radix ( Astragalus ) , the root of Astragalus membranaceus Bunge , has been widely applied to treat patients with viral diseases , including viral myocarditis in China . The present study was designed to evaluate the protective effects of Astragalus on the function of sarcoplasmic reticulum calcium ATPase ( P16615 ) activity and endothelin system at acute and chronic periods of myocarditis mice induced by CVB(3) infection . Astragalus feeding ( 2.2 mg/kg/day ) could significantly increase the survival rate , alleviate pathological alterations and serum cardiac troponin I ( cTnI ) , as well as restore impaired SERCA activity at the acute stage . Low affinity and capacity of ETR were reversed with Astragalus after the first CVB(3) inoculation up to 7 days and after the second virus inoculation up to 150 days . In the meantime , the contents of cardiac ET-1 and P01160 were reduced . Comparison the myocarditis mice treated with DB00790 ( 0.44 mg/kg/day ) , an P12821 inhibitor , shows that Astragalus achieved a similar effect on survival rate , P16615 and ET system . These results indicated that the beneficial effects of Astragalus and DB00790 for treating viral myocarditis might be partly mediated by preserving the functions of SERCA 2 activity and ET system . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Soluble epoxide hydrolase inhibitor , TUPS , protects against isoprenaline-induced cardiac hypertrophy . BACKGROUND AND PURPOSE : We have previously shown that isoprenaline-induced cardiac hypertrophy causes significant changes in the expression of cytochromes P450 ( CYP ) and soluble epoxide hydrolase ( sEH ) genes . Therefore , it is important to examine whether the inhibition of sEH by 1-(1-methanesulfonyl-piperidin-4-yl)-3-(4-trifluoromethoxy-phenyl)-urea ( TUPS ) will protect against isoprenaline-induced cardiac hypertrophy . EXPERIMENTAL APPROACH : Male Sprague-Dawley rats were treated with TUPS ( 0.65 mg kg(-1) day(-1) , p.o. ) , isoprenaline ( 5 mg kg(-1) day(-1) , i.p. ) or the combination of both . In vitro H9c2 cells were treated with isoprenaline ( 100 μM ) in the presence and absence of either TUPS ( 1 μM ) or 11,12 EET ( 1 μM ) . The expression of hypertrophic , fibrotic markers and different CYP genes were determined by real-time PCR . KEY RESULTS : Isoprenaline significantly induced the hypertrophic , fibrotic markers as well as the heart to body weight ratio , which was significantly reversed by TUPS . Isoprenaline also caused an induction of P04798 , Q16678 , CYP2B1 , CYP2B2 , CYP4A3 and CYP4F4 gene expression and TUPS significantly inhibited this isoprenaline-mediated effect . Moreover , isoprenaline significantly reduced 5,6- , 8,9- , 11,12- and 14,15-EET and increased their corresponding 8,9- , 11,12- and 14,15-dihydroxyeicosatrienoic acid ( DHET ) and the 20-HETE metabolites . TUPS abolished these isoprenaline-mediated changes in arachidonic acid ( AA ) metabolites . In H9c2 cells , isoprenaline caused a significant induction of P01160 , DB04899 and P34913 mRNA levels . Both TUPS and 11,12-EET significantly decreased this isoprenaline-mediated induction of P01160 , DB04899 and P34913 . CONCLUSIONS AND IMPLICATIONS : TUPS partially protects against isoprenaline-induced cardiac hypertrophy , which confirms the role of sEH and CYP enzymes in the development of cardiac hypertrophy . Analgesic and Anti-Inflammatory Activities of Methanol Extract of Cissus repens in Mice . The aim of this study was to investigate possible analgesic and anti-inflammatory mechanisms of the CR(MeOH) . Analgesic effect was evaluated in two models including acetic acid-induced writhing response and formalin-induced paw licking . The anti-inflammatory effect was evaluated by λ-carrageenan-induced mouse paw edema and histopathologic analyses . The results showed that CR(MeOH) ( 500 mg/kg ) decreased writhing response in the acetic acid assay and licking time in the formalin test . CR(MeOH) ( 100 and 500 mg/kg ) significantly decreased edema paw volume at 4th to 5th hours after λ-carrageenan had been injected . Histopathologically , CR(MeOH) abated the level of tissue destruction and swelling of the edema paws . These results were indicated that anti-inflammatory mechanism of CR(MeOH) may be due to declined levels of NO and MDA in the edema paw through increasing the activities of SOD , GPx , and GRd in the liver . Additionally , CR(MeOH) also decreased IL-1β , P05231 , NFκB , P01375 -α , P35354 , and P35228 levels . The contents of two active ingredients , ursolic acid and lupeol , were quantitatively determined . This paper demonstrated possible mechanisms for the analgesic and anti-inflammatory effects of CR(MeOH) and provided evidence for the classical treatment of Cissus repens in inflammatory diseases . Bayesian meta-analysis of tissue angiotensin-converting enzyme inhibitors for reduction of adverse cardiovascular events in patients with diabetes mellitus and preserved left ventricular function . The role of angiotensin-converting enzyme ( P12821 ) inhibitors in diabetic patients with preserved ventricular function is uncertain . Tissue P12821 inhibitors have been defined by increased lipophilicity and structural characteristics that result in greater tissue-specific P12821 binding when compared with plasma P12821 inhibitors . A Bayesian meta-analysis of randomized trials was conducted to evaluate tissue P12821 inhibitors in prevention of cardiovascular disease among patients with diabetes mellitus and preserved left ventricular function . Four trials were selected that evaluated 2 different P12821 inhibitors and included 10,328 patients ( 43,517 patient-years ) . The DB00790 Substudy in Coronary Artery Disease and Diabetes ( PERSUADE ) and the DB00790 Protection Against Recurrent Stroke Study ( PROGRESS ) compared the effects of perindopril vs a placebo , and the Heart Outcomes Prevention Evaluation ( HOPE ) and the Non- P01308 -Dependent Diabetes , Hypertension , Microalbuminuria , Proteinuria , Cardiovascular Events , and Ramipril ( DIABHYCAR ) study investigated the impact of ramipril vs a placebo . Bayesian meta-analysis of sequential trials and sensitivity analysis of therapeutic response were subsequently computed . Bayesian meta-analysis determined reduced risk of cardiovascular mortality ( PB=.991 ) , myocardial infarction ( PB=.999 ) , and the need for invasive coronary revascularization ( PB=.995 ) when compared with placebo . Total mortality was also decreased ( PB=.967 ) , while the risk of stroke ( PB=.907 ) and hospitalization for heart failure ( PB=.923 ) were impacted . Bayesian meta-analysis of randomized trials suggests that tissue P12821 inhibitors decrease the probability that diabetic patients with preserved left ventricular function will experience myocardial infarctions and cardiovascular death and reduce overall mortality . Inhibition of central angiotensin converting enzyme ameliorates scopolamine induced memory impairment in mice : role of cholinergic neurotransmission , cerebral blood flow and brain energy metabolism . Evidences indicate that inhibition of central P00797 angiotensin system ( DB01367 ) ameliorates memory impairment in animals and humans . Earlier we have reported involvement of central angiotensin converting enzyme ( P12821 ) in streptozotocin induced neurodegeneration and memory impairment . The present study investigated the role of central P12821 in cholinergic neurotransmission , brain energy metabolism and cerebral blood flow ( Q03701 ) in model of memory impairment induced by injection of scopolamine in mice . DB00790 ( 0.05 and 0.1 mg/kg , PO ) was given orally for one week before administration of scopolamine ( 3mg/kg , IP ) . Then , memory function was evaluated by Morris water maze and passive avoidance tests . Q03701 was measured by laser Doppler flowmetry . Biochemical and molecular parameters were estimated after the completion of behavioral studies . DB00747 caused impairment in memory which was associated with reduced Q03701 , acetylcholine ( ACh ) level and elevated acetylcholinesterase ( P22303 ) activity and malondialdehyde ( MDA ) level . DB00790 ameliorated scopolamine induced amnesia in both the behavioral paradigms . Further , perindopril prevented elevation of P22303 and MDA level in mice brain . There was a significant increase in Q03701 and ACh level in perindopril treated mice . However , scopolamine had no significant effect on DB00171 level and mRNA expression of angiotensin receptors and P12821 in cortex and hippocampus . But , perindopril significantly decreased P12821 activity in brain without affecting its mRNA expression . The study clearly showed the interaction between P12821 and cholinergic neurotransmission and beneficial effect of perindopril can be attributed to improvement in central cholinergic neurotransmission and Q03701 .

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Glomerular mRNA expression of prothrombotic and antithrombotic factors in renal transplants with thrombotic microangiopathy . BACKGROUND : Thrombotic microangiopathy ( TMA ) in renal transplants ( rTx-TMA ) is a serious complication and is usually either recurrent TMA ( RecTMA ) due to humoral rejection ( HR-TMA ) or due to calcineurin inhibitor toxicity ( CNI-TMA ) . Although the triggers are known , our knowledge about the thrombogenic transcriptome changes in the microvessels is rudimentary . METHODS : We examined the expression of several prothrombotic and antithrombotic genes in 25 biopsies with rTx-TMA ( 6 RecTMA , 9 HR-TMA , and 10 CNI-TMA ) and 8 controls . RNA from microdissected glomeruli of paraffin-embedded tissue was isolated and mRNA transcripts were quantified with real-time polymerase chain reaction after preamplification . Results were correlated with clinicopathologic parameters . RESULTS : Glomerular mRNA expression of Q9Y5W3 , O43474 , and tPA was lower and that of P05121 was higher in rTx-TMA than in the controls . Glomerular mRNA expression of Q9Y5W3 and O43474 correlated with that of tPA and inversely with that of P05121 in rTx-TMA . The mRNA expression of complement regulators P15529 and P13987 were higher in rTx-TMA than in the controls . Only in HR-TMA were glomerular Q76LX8 and P08174 down-regulated . CONCLUSIONS : The glomerular capillary bed seems to contribute to all subtypes of rTx-TMA by down-regulation of the endothelial transcription factors Q9Y5W3 and O43474 , indicating dedifferentiation with subsequent up-regulation of P05121 and down-regulation of tPA , resulting in inhibition of local fibrinolysis . Decreased glomerular expression of Q76LX8 and P08174 could be an additional pathway toward microthrombosis exclusively in HR-TMA . Allogeneic stem cell transplantation using alemtuzumab-containing regimens in severe aplastic anemia . DB00087 , a humanized anti- P31358 , IgG1 monoclonal antibody , is used to reduce graft-versus- host disease ( GVHD ) and aid engraftment after allogeneic haemopoietic stem cell transplant ( HSCT ) . Its associated low incidence of GVHD makes it an attractive alternative to anti-thymocyte globulin ( ATG ) in transplant conditioning regimen for severe aplastic anaemia ( P0DJI8 ) . We have reviewed the use of alemtuzumab-based conditioning regimen for HSCT in P0DJI8 and show that it results in sustained haematological engraftment , a very low incidence of chronic GVHD without an increase in viral infections . Intriguingly , alemtuzumab appears to induce tolerance post-HSCT with the findings of stable mixed T cell chimerism with full donor myeloid chimerism and the absence of chronic GVHD , and which persist on withdrawal of post-graft immunosuppression . Finally , its low toxicity profile may permit future application of HSCT to older patients with P0DJI8 who fail to respond to immunosuppressive therapy . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Immunity 12 years after alemtuzumab in RA : CD5⁺ B-cell depletion , thymus-dependent T-cell reconstitution and normal vaccine responses . OBJECTIVES : Lymphocyte depleting therapies have been used to treat refractory autoimmune disease , including RA , but treatment may be associated with long-term lymphopenia . It is unclear whether delayed reconstitution preferentially affects lymphocyte subsets , how this modulates immune challenges and whether thymic function influences the outcome . These questions are now addressed in a detailed analysis of RA patients 12 years after alemtuzumab ( anti- P31358 ) treatment . METHODS : Blood was obtained from 20 RA patients 12 years after alemtuzumab treatment . Lymphocyte subsets were enumerated by flow cytometry . T-cell receptor excision circles ( TRECs ) /ml were determined to quantify thymic function , and serological responses to neoantigens and recall antigens were assessed . RESULTS : RA patients remained lymphopenic 12 years after their first dose of alemtuzumab . P06127 (+) B cells , which may be associated with autoantibody production , were significantly reduced in alemtuzumab-treated patients compared with age-matched disease controls . In addition , naïve and memory P01730 (+) T-cell subsets were present in altered proportions in patients who had received alemtuzumab , with increased effector memory P01730 (+) T cells , and decreased naïve and central memory P01730 (+) T cells . TRECs were detectable in alemtuzumab-treated patients and correlated with P01730 (+) lymphocyte counts . Vaccine responses to neoantigens and recall antigens fell within the normal range for an ageing population . CONCLUSIONS : DB00087 therapy resulted in long-term alterations in lymphocyte subsets . The significance of these changes remains uncertain but patients respond normally to antigenic challenges . Thymic function remains an important determinant of T-cell reconstitution even several years after lymphocytotoxic therapy . Concomitant targeting of tumor cells and induction of T-cell response synergizes to effectively inhibit trastuzumab-resistant breast cancer . DB00072 is an iconic rationally designed targeted therapy for P04626 -positive breast cancers . However , the low response rate and development of resistance call for novel approaches for the treatment of patients . Here , we report that concurrent targeting of tumor cells and activation of T cells in the tumor microenvironment results in a synergistic inhibitory effect on tumor growth and overcomes resistance in two distinct P60484 loss-mediated trastuzumab-resistant mammary tumor mouse models . In vivo combination treatment with P04626 /Neu antibody and Akt inhibitor triciribine effectively inhibited tumor growth in both models via inhibiting PI3K/AKT and mitogen-activated protein kinase signaling accompanied by increased T-cell infiltration in the tumor microenvironment . We showed that both CD8(+) and P01730 (+) T cells were essential to the optimal antitumor effect of this combination treatment in an IFN-γ-dependent manner . Importantly , the antitumor activities of P04626 /Neu antibody and triciribine combination treatment were further improved when coinhibitory receptor cytotoxic T-lymphocyte-associated antigen 4 was blocked to enhance the T-cell response . Our data indicate that multitargeted combinatorial therapies targeting tumor cells and concomitantly enhancing T-cell response in the tumor microenvironment could cooperate to exert maximal therapeutic activity , suggesting a promising clinical strategy for treating trastuzumab-resistant breast cancers and other advanced malignancies . Association between high interleukin-6 levels and adverse outcome after autologous haemopoietic stem cell transplantation . We studied interleukin-6 ( P05231 ) levels on the day of transplantation in 31 patients undergoing autologous haemopoietic stem cell transplantation ( P09683 ) ( either peripheral blood stem cell transplantation ( PBSCT ) or bone marrow transplantation ( BMT ) ) for neoplastic diseases to determine if there was a relationship between P05231 level and rate of haemopoietic recovery , length of stay in hospital , and survival . There was no apparent delay in post-transplant recovery associated with elevated P05231 levels . However , increased values of P05231 tended to be associated with an increased length of stay in hospital ( P = 0.083 ) . There was a highly significant adverse association between higher P05231 levels and survival following transplantation ( P = 0.0001 ) . This association remained significant ( P = 0.013 ) in the uniform subgroup of patients with malignant lymphoma with chemosensitive disease who had undergone BMT ( that is , excluding patients who had undergone PBSCT ) ( n = 13 ) . Knowledge of P05231 levels on the day of transplant has the potential to provide valuable prognostic information in patients undergoing autologous haemopoietic P09683 . Pharmacokinetics of alemtuzumab used for in vivo and in vitro T-cell depletion in allogeneic transplantations : relevance for early adoptive immunotherapy and infectious complications . Persistence of alemtuzumab at lympholytic concentrations after reduced-intensity conditioning allogeneic stem cell transplantations ( RITs ) could impair immune reconstitution and reduce donor T-cell-mediated graft-versus-leukemia/lymphoma ( GVL ) effects , derived from the graft or subsequent adoptive immunotherapy . We have studied the pharmacokinetics of alemtuzumab in 2 different groups : Q92963 ( 100 mg alemtuzumab in vivo over 5 days ) and myeloablative allografts ( 20 mg alemtuzumab added in vitro to the stem cells prior to return ) . DB00087 concentrations in Q92963 patients were in excess of that required to kill infused donor P31358 + cells at the time of transplantation and remained at potentially lympholytic levels ( > 0.1 microg/mL ) for approximately 56 days after transplantation , 26 days longer than for the myeloablative group . Total lymphocyte counts were significantly lower in the Q92963 group persisting beyond 6 months after transplantation ( P =.005 ) , and median absolute P01730 counts higher than 200 x 106/L were delayed until 9 months after transplantation . In vivo alemtuzumab enables haploidentical human leukocyte antigen-mismatched hematopoietic stem-cell transplantation without ex vivo graft manipulation . BACKGROUND : DB00087 , a humanized monoclonal antibody directed against human P31358 , has a strong lympholytic effect . This study evaluates the safety of unmanipulated peripheral blood stem-cell transplantation from two or three loci-mismatched related donors using alemtuzumab in vivo . METHODS : A total body irradiation-based regimen was used in young patients , whereas those 50 years or older received fludarabine-based conditioning . DB00087 was added to these regimens by intravenous infusion at 0.2 mg/kg per day for 6 days ( days -8 to -3 ) . RESULTS : We treated 12 patients with a median age of 49.5 years . Eight patients demonstrated active disease , and four patients demonstrated acute leukemia in high-risk remission . All achieved neutrophil engraftment a median of 17.5 days after transplantation with complete donor-type chimerism . The cumulative incidence of grades III to IV acute graft-versus-host disease was only 9 % . Infection-related deaths were not observed . CD3+/ P01730 + and CD3+/CD8+ T cells were strongly suppressed within 2 months after transplantation , but recovered on day 90 . Relapse was observed in five of eight patients who underwent transplantation for active disease , whereas none of the three patients who underwent transplantation in first remission had a relapse . CONCLUSIONS : We conclude that in vivo alemtuzumab enables haploidentical hematopoietic stem-cell transplantation without ex vivo graft manipulation . Gene transfer of GM- P04141 , P33681 and CD154 cDNA enhances survival in a murine model of acute leukemia with persistence of a minimal residual disease . Gene transfer of various cytokines and co-stimulatory molecules has been reported to induce a potent antileukemic immunity in murine models , however , the relative efficiency and possible synergistic effects between candidate genes have not been extensively investigated . We analyzed in a murine model of P11274 / P00519 acute leukemia whether gene transfer of CD154 , P33681 or GM- P04141 as a single agent or combination of CD154 + GM- P04141 , P33681 + CD154 and GM- P04141 + P33681 in leukemic cells could enhance survival . We observed that CD154 gene transfer induced a marked inhibition of leukemogenicity , and also that CD154 and combination of GM- P04141 and P33681 gene transfer protected mice against subsequent challenge with leukemic cells and had a therapeutic effect for a pre-established leukemia disease . We also found minimal residual leukemic disease by RT-PCR for 6 to 12 months in 0 to 25 % of animals injected with transduced leukemic cells and surviving the challenge without evidence of disease , except in the control empty plasmid group where very few mice survived the challenge but all of those were positive by RT-PCR . These findings suggest that leukemic cell vaccination by gene transfer can induce a tumor dormancy phenomenon compatible with long-term survival . Expression of membrane complement regulators , P15529 , P08174 and P13987 , in mesothelial cells of patients on peritoneal dialysis therapy . We investigated the expression of membrane complement regulators ( CRegs ) , P15529 , P08174 and P13987 in human mesothelial cells , and correlated with clinical background and level of complement ( C ) activation products in peritoneal dialysis ( PD ) fluids ( PDF ) to clarify influence of the C activation system in PD patients . Expression of CRegs was assessed on primary cultures of mesothelial cells ( HPMC ) harvested from PD fluid of 31 PD patients . Because expression of P08174 but not P15529 and P13987 in mesothelial cells was significantly correlated to value of dialysate-to-plasma creatinine concentration ratio ( D/P Cre ) ( p < 0.005 ) as an indicator of peritoneal function , we focused on analysis of P08174 expression of HPMCs in comparison with levels of C activation products in the PDF of the PD patients , and their background factors . When comparing expression of the CRegs between systemic neutrophils and HPMC , no correlation was observed , supporting that change of CRegs ' expression in HPMC was independently occurring in the peritoneum . Expression of P08174 protein in HPMC was closely correlated with expression at the mRNA level ( p < 0.0001 ) and was inversely correlated with levels of sC5b-9 ( p < 0.05 ) , but not P01024 , C4 , P05231 and Q8WXI7 in the PDF . Complications of diabetes , usage of icodextrin and residual renal function were not correlated with change of P08174 expression in HPMCs . Our data show that the process of PD therapy modifies expression of P08174 on peritoneal mesothelium and triggers local C activation . These findings support efforts to modify PD therapy to limit effects on activation and regulation of the C system . P11021 / P11021 / P11021 /Dna K is a universal therapeutic target for human disease . The chaperone P11021 /Dna K is conserved throughout evolution down to prokaryotes . The P11021 inhibitor OSU-03012 ( AR-12 ) interacted with sildenafil ( Viagra ) or tadalafil ( DB00820 ) to rapidly reduce P11021 levels in eukaryotes and as a single agent reduce Dna K levels in prokaryotes . Similar data with the drug combination were obtained for : HSP70 , HSP90 , P14625 , P30101 , HSP27 , P25685 and HSP60 . OSU-03012/sildenafil treatment killed brain cancer stem cells and decreased the expression of : O15118 and TIM1 ; P11279 ; and NTCP1 , receptors for Ebola/Marburg/Hepatitis A , Lassa fever , and Hepatitis B viruses , respectively . Pre-treatment with OSU-03012/sildenafil reduced expression of the coxsakie and adenovirus receptor in parallel with it also reducing the ability of a serotype 5 adenovirus or coxsakie virus B4 to infect and to reproduce . Similar data were obtained using Chikungunya , Mumps , Measles , Rubella , RSV , CMV , and Influenza viruses . OSU-03012 as a single agent at clinically relevant concentrations killed laboratory generated antibiotic resistant E. coli and clinical isolate multi-drug resistant N. gonorrhoeae and MRSE which was in bacteria associated with reduced Dna K and Q9UIJ5 A expression . The O76074 inhibitors sildenafil or tadalafil enhanced OSU-03012 killing in N. gonorrhoeae and MRSE and low marginally toxic doses of OSU-03012 could restore bacterial sensitivity in N. gonorrhoeae to multiple antibiotics . Thus , Dna K and bacterial phosphodiesterases are novel antibiotic targets , and inhibition of P11021 is of therapeutic utility for cancer and also for bacterial and viral infections . New antibody conjugates in cancer therapy . Targeting of radiation , drugs , and protein toxins to cancers selectively with monoclonal antibodies ( MAbs ) has been a topic of considerable interest and an area of continued development . Radioimmunotherapy ( RAIT ) of lymphoma using directly labeled MAbs is of current interest after approval of two radiolabeled anti- P11836 MAbs , as illustrated with the near 100 % overall response rate obtained in a recent clinical trial using an investigational radiolabeled anti- P20273 MAb , 90Y-epratuzumab . The advantage of pretargeted RAIT over directly labeled MAbs is continuing to be validated in preclinical models of lymphoma and solid tumors . Importantly , the advantages of combining RAIT with radiation sensitizers , with immunotherapy , or a drug conjugate targeting a different antigen are being studied clinically and preclinically . The area of drug-conjugated antibodies is progressing with encouraging data published for the trastuzumab-DM1 conjugate in a phase I clinical trial in P04626 -positive breast cancer . The Dock-and-Lock platform technology has contributed to the design and the evaluation of complex antibody-cytokine and antibody-toxin conjugates . This review describes the advances made in these areas , with illustrations taken from advances made in the authors ' institutions . Bloodstream infections in organ transplant recipients receiving alemtuzumab : no evidence of occurrence of organisms typically associated with profound T cell depletion . BACKGROUND : DB00087 is a humanized monoclonal antibody directed against P31358 , a cell surface antigen expressed on B and T lymphocytes , monocytes and NK cells . Its use results in a profound decrease in P01730 positive T lymphocytes . DB00087 is used as induction immunosuppression and therapy for rejection in organ transplant recipients in some centers . We followed a cohort of 449 consecutive transplant recipients who received alemtuzumab to determine the occurrence of bloodstream infections , particularly those previously associated with decrease in P01730 positive T lymphocytes . PATIENTS AND METHODS : Fifteen percent ( 69/449 ) patients had at least one episode of bloodstream infection . However , no patient had bacteremia with Streptococcus pneumoniae , Listeria monocytogenes , non-typhoidal Salmonella or Mycobacterium avium complex . Fungaemia was rare , occurring in 1.5 % of patients . The most common organisms isolated from the blood were Staphylococcus aureus ( 21 episodes ) , coagulase negative Staphylococcus ( 14 episodes ) , Klebsiella pneumoniae ( 12 episodes ) , Enterococcus faecium ( 11 episodes ) , Pseudomonas aeruginosa ( 10 episodes ) , Enterococcus faecalis ( 9 episodes ) and Escherichia coli ( 7 episodes ) . DISCUSSION : We conclude that although alemtuzumab use is associated with profound P01730 positive T lymphocyte depletion , alemtuzumab does not seem to be associated with an increased risk of bloodstream infection with pathogens typically seen in other disorders of P01730 cell depletion , such as acquired immunodeficiency syndrome . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . DB00087 use in relapsed and refractory chronic lymphocytic leukemia : a history and discussion of future rational use . In this review , we outline the clinical experience with single-agent alemtuzumab as a treatment for relapsed and refractory chronic lymphocytic leukemia ( CLL ) in both prospective and retrospective trials and describe the multiagent use of the drug with the goal of updating clinicians on recent developments and possible future rational combinations . DB00087 , an antibody targeting the lymphocyte-specific surface marker P31358 , is an approved agent for the treatment of CLL . Despite its demonstrated efficacy , likely secondary to concerns regarding infectious complications , it is most commonly used in the relapsed and refractory setting . Given alemtuzumab 's unique mechanism of action it has been demonstrated to have activity in disease that is refractory to both alkylating agents and purine analogs . Furthermore , it has activity in P04637 -mutated disease , which has the worst prognosis of any subset of CLL . DB00087 has greater efficacy on circulating disease relative to nodal disease . Rational combinations are attempting to use these attributes to increase response rates in patients with relapsed and refractory disease . Inhibitory effect of fluvastatin , an P04035 inhibitor , on the expression of adhesion molecules on human monocyte cell line . The effect of fluvastatin , an P04035 inhibitor , was investigated on the adhesive interaction between U937 cells , the human monocyte cell line , and human umbilical vein endothelial cells ( HUVEC ) , focusing on the expression of adhesion molecules . U937 treated with fluvastatin lowered the capacity for binding to HUVEC . DB01095 at 0.1 microM or more inhibited the expression of lymphocyte function associated antigen-1 ( LFA-1 ) on U937 and intercellular adhesion molecule-1 ( P05362 ) on U937 . The expression of P05362 on HUVEC was not inhibited by fluvastatin . The inhibitory effects of fluvastatin on the expression of adhesion molecules on U937 were completely reversed by the addition of mevalonate . Because fluvastatin did not affect the expression of other cell surface markers , P01730 and CD71 , the inhibitory effects of fluvastatin on adhesion molecule expression could not be attributed to the non-specific suppression of the cell . It is conceivable that cellular interaction between monocytes and endothelial cells is inhibited by fluvastatin , mediated via reducing the expression of adhesion molecules , particularly in the side of monocyte . Dissecting the complement pathway in hepatic injury and regeneration with a novel protective strategy . Liver resection is commonly performed under ischemic conditions , resulting in two types of insult to the remnant liver : ischemia reperfusion injury ( IRI ) and loss of liver mass . Complement inhibition is recognized as a potential therapeutic modality for IRI , but early complement activation products are also essential for liver regeneration . We describe a novel site-targeted murine complement inhibitor , P20023 - P13987 , which specifically inhibits the terminal membrane attack complex ( MAC ) , and we use this protein to investigate the complement-dependent balance between liver injury and regeneration in a clinical setting of pharmacological inhibition . P20023 - P13987 did not impact in vivo generation of P01024 and P01031 activation products but was as effective as the P01024 activation inhibitor P20023 -Crry at ameliorating hepatic IRI , indicating that the MAC is the principle mediator of hepatic IRI . Furthermore , unlike P01024 or P01031 inhibition , P20023 - P13987 was not only protective but significantly enhanced hepatocyte proliferation after partial hepatectomy , including when combined with ischemia and reperfusion . Remarkably , P20023 - P13987 also enhanced regeneration after 90 % hepatectomy and improved long-term survival from 0 to 70 % . P20023 - P13987 functioned by increasing hepatic P01375 and P05231 levels with associated P40763 and Akt activation , and by preventing mitochondrial depolarization and allowing recovery of DB00171 stores . Cucurbitacin B induced Q13315 -mediated DNA damage causes G2/M cell cycle arrest in a ROS-dependent manner . Cucurbitacins are a class of triterpenoids widely distributed in plant kingdom with potent anti-cancer activities both in vitro and in vivo by inducing cycle arrest , autophagy , and apoptosis . Cucurbitacin B ( Cuc B ) , could induce S or G2/M cell cycle arrest in cancer cells while the detailed mechanisms remain to be clear . This study was designed to precisely dissect the signaling pathway(s) responsible for Cuc B induced cell cycle arrest in human lung adenocarcinoma epithelial A549 cells . We demonstrated that low concentrations of Cuc B dramatically induced G2/M phase arrest in A549 cells . Cuc B treatment caused DNA double-strand breaks ( DSBs ) without affecting the signal transducer and activator of transcription 3 ( P40763 ) , the potential molecular target for Cuc B . Cuc B triggers Q13315 -activated Chk1-Cdc25C-Cdk1 , which could be reversed by both Q13315 siRNA and Chk1 siRNA . Cuc B also triggers Q13315 -activated p53-14-3-3-σ pathways , which could be reversed by Q13315 siRNA . Cuc B treatment also led to increased intracellular reactive oxygen species ( ROS ) formation , which was inhibited by N-acetyl-l-cysteine ( Q9C000 ) pretreatment . Furthermore , Q9C000 pretreatment inhibited Cuc B induced DNA damage and G2/M phase arrest . Taken together , these results suggested that Cuc B induces DNA damage in A549 cells mediated by increasing intracellular ROS formation , which lead to G2/M cell phase arrest through Q13315 -activated Chk1-Cdc25C-Cdk1 and p53-14-3-3-σ parallel branches . These observations provide novel mechanisms and potential targets for better understanding of the anti-cancer mechanisms of cucurbitacins . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . DB00087 in the treatment of chronic lymphocytic lymphoma . DB00087 was the first monoclonal antibody to be humanized , a process which embeds rodent sequence fragments in a human IgG framework . The antibody target is P31358 , an antigen expressed on normal lymphocytes as well as many T- and B-cell neoplasms . It therefore has a potential broad application across a spectrum of B- and T-cell malignancies as well as use as an immunosuppressant drug in , for example , bone marrow transplantation . The original licensing in the USA and Europe was for the treatment of fludarabine-refractory chronic lymphocytic leukemia ( CLL ) . However , recent trials using alemtuzumab as a first-line agent for CLL have shown superior response rates compared with traditional alkylator therapy and this has led to US FDA approval for first-line treatment for CLL . It seems to be particularly useful in patients with CLL who have deletion of the P04637 tumor suppressor gene , a subset of disease that responds poorly to other currently available chemotherapeutics . Targetting esophageal and gastric cancers with monoclonal antibodies . Target therapies and notably monoclonal antibodies are currently being considered for esophageal , gastric , and gastroesophageal junction cancers . P00533 was found to be overexpressed in 60-86 % of gastric or gastroesophageal tumors and in 50-70 % of esophageal cancers . Cetuximab was shown to be a radiosensitizing agent in the treatment of ENT neoplasia . These results led to several phase II encouraging therapeutic trials evaluating the combination of cetuximab with radiochemotherapy in locally advanced esophageal cancers . Numerous encouraging phase II trials evaluating cetuximab combined with chemotherapy in patients with gastric adenocarcinoma or gastroesophageal junction cancer were reported . These promising results are still to be confirmed by the ongoing phase III trials . Several studies reported P04626 overexpression in gastric cancer ( 7-34 % ) , which appeared to be associated with poorer prognosis . DB00072 is a monoclonal antibody directed against the extracellular P04626 domain . The international phase III trial known as ToGA ( DB00072 for Gastric Cancer ) aimed to determine the clinical efficacy and acceptable toxicity profile of trastuzumab in combination with first-line chemotherapy in P04626 -overexpressing gastric or gastroesophageal cancer . Angiogenesis is an essential step in the initial phase of tumorigenesis , and it is normally absent from healthy tissues except for particular physiological situations , such as wound healing . P15692 plays a role in endothelial growth and angiogenesis . DB00112 , a humanized monoclonal anti- P15692 antibody , is currently being studied for gastric cancer . The phase III AVAGAST study , evaluating bevacizumab in association with chemotherapy in advanced gastric adenocarcinoma , did not achieve its primary aim of improved OS in bevacizumab-treated patients . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . P03372 -immunoreactive neurons contain calcitonin gene-related peptide , methionine-enkephalin or tyrosine hydroxylase in the female rat preoptic area . We have shown in our previous studies that estrogen treatment selectively influences calcitonin gene-related peptide ( P80511 ) - , methionine-enkephalin ( DB00134 -Enk ) - and tyrosine hydroxylase ( TH ) -immunoreactive ( IR ) intensities in the neurons of the periventricular preoptic nucleus ( Q9H237 ) and the medial preoptic area ( DB00603 ) of the female rat . In the present study , we examined whether estrogen receptor ( ER ) -IR neurons in the Q9H237 and DB00603 contain P80511 , DB00134 -Enk , or TH using a double-labeling immunohistochemical method and investigated changes in the number of double-labeling cells upon treatment with estrogen . Brain sections of ovariectomized rats and ovariectomized and estrogen-treated rat were stained using the avidin-biotin-peroxidase complex method followed by the peroxidase-anti-peroxidase method . The sections were first incubated with an anti-ER antibody in conjunction with nickel diaminobenzidine which produces a dark blue reaction product in the nucleus . Subsequently , P80511 , DB00134 -Enk or TH antisera were applied to these sections and the resulting brown diaminobenzidine reaction product in the cytoplasm was examined . Neurons that were double-labeled for ER and P80511 , DB00134 -Enk or TH were investigated in the Q9H237 and DB00603 . The number of doubly labeled ER/ P80511 - and ER/TH-IR neurons was large , whereas the number of ER/ DB00134 -Enk-IR neurons was small . These results suggest that ER in the Q9H237 and DB00603 may be more closely related to the mechanism of changes in P80511 - and TH-IR intensities upon estrogen treatment than that in DB00134 -Enk-IR intensity . Novel and emerging drugs for rarer chronic lymphoid leukaemias . Rarer chronic lymphoid leukaemias represent a challenge to the clinicians due to the limited information on their pathogenesis , difficulties on setting up prospective clinical trials and to their refractoriness to drugs used in the most common form of chronic lymphocytic leukaemia ( CLL ) . In this review all these issues are addressed in three B-cell leukaemias : B-cell prolymphocytic leukaemia ( B-PLL ) , hairy cell leukaemia ( HCL ) and HCL-variant and three T-cell leukaemias : T-cell prolymphocytic leukaemia ( T-PLL ) , T-cell large granular lymphocytic leukaemia ( T-cell LGLL ) and adult T-cell leukaemia lymphoma ( ATLL ) . Data will be presented on the natural history , current therapies and emerging drugs potentially useful in the treatment of patients with these leukaemias . Emphasis is made on : 1- the novel agents targeting a variety of B and T-cell antigens expressed on the surface of the leukaemic cells ; these are either unconjugated monoclonal antibodies ( McAb ) such as DB00073 ( anti- P11836 ) , the second and third generation of anti- P11836 McAbs , DB00087 ( anti- P31358 ) , Siplizumab ( anti- P06729 ) , DB00111 ( anti-CD25 ) and KW-0761 , an anti-chemokine receptor 4 ( CCR4 ) or McAbs conjugated to toxins such as P20273 linked to the pseudomonas exotoxin or radiolabelled McAb ; 2- the use of new purine nucleosides such as nelarabine and 3- agents targeting deregulated genes in the leukaemic cells from these diseases such as the Poly ( ADP-ribose ) polymerase ( PARP ) Olarapib in T-PLL with deregulation of the ataxia telangiectasia mutated ( Q13315 ) gene . Data of phase I and II clinical studies with these agents as well as the potential and current use of other drugs are outlined . DB00087 in the treatment of refractory acute rejection and bronchiolitis obliterans syndrome after human lung transplantation . Despite substantial improvements in early survival after lung transplantation , refractory acute rejection ( RAR ) and bronchiolitis obliterans syndrome ( BOS ) remain major contributors to transplant-related morbidity and mortality . We have utilized alemtuzumab , a humanized anti- P31358 antibody which results in potent lymphocyte depletion , in consecutive patients with RAR ( n = 12 ) or BOS ( n = 10 ) . All patients failed conventional treatment with methylprednisolone and antithymocyte globulin and received strict infection prophylaxis . DB00087 significantly improved histological rejection scores in RAR . Total rejection grade/biopsy was 1.98 +/- 0.25 preceding alemtuzumab versus 0.33 +/- 0.14 posttreatment , p-value < 0.0001 ( with a similar number of biopsies/patient per respective time interval ) . Freedom from BOS was observed in 65 % of RAR patients 2 years after alemtuzumab treatment . Although there was no statistically significant change in forced expiratory volume in 1 second ( FEV1 ) before and after alemtuzumab treatment in patients with BOS , a stabilization or improvement in BOS grade occurred in 70 % of patients . Patient survival 2 years after alemtuzumab for BOS was 69 % . Despite a dramatic decline in P01730 counts in alemtuzumab-treated patients , only one patient developed a lethal infection . Thus , we provide the first evidence that alemtuzumab is a potentially useful therapy in lung transplant recipients with RAR or BOS . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . P80511 expression increases in the ventral horn rostral to spinal cord transection . Rats underwent spinal cord transection ( P09683 ) at the mid-thoracic level , and calcitonin gene-related peptide-like immunoreactivity ( P80511 -LI ) was analysed in the ventral horn rostral and caudal to the injury . P80511 expression decreased in motor neurones caudal to the lesion , as previously described . Rostral to the transection , however , the area of P80511 -LI increased in the ventral horn 1 and 2 weeks following P09683 , although the number of neurones expressing P80511 did not differ from controls . Immunofluorescent fibres were seen in the ventral horn rostral to the lesion only after P09683 . Thus , we report the novel observation that P80511 expression in motor neurones is upregulated rostral to a spinal cord lesion , perhaps due to an imbalance of descending and intraspinal inputs . Preimplantation genetic diagnosis for cancer predisposition syndromes . OBJECTIVES : Mutations in the P25054 , P35240 and P38398 genes cause adult-onset cancer predisposition syndromes . Prenatal diagnosis ( P01160 ) and selective pregnancy termination for adult-onset disorders is emotionally difficult and , in some cases , socially not well accepted . Preimplantation genetic diagnosis ( P52209 ) appears as an attractive alternative to P01160 , as it ensures the establishment of a pregnancy free of the mutation from the onset , circumventing the potentially difficult decision of termination of pregnancy . METHODS : Development of single-cell PCRs using Epstein-Barr virus transformed lymphoblasts as single-cell model , followed by clinical application in P52209 . RESULTS : A total of five duplex-PCRs were developed , three for adenomatous polyposis of the colon ( P25054 ) , one for neurofibromatosis type 2 ( P35240 ) and one for inherited breast and ovarian cancer caused by P38398 mutations . Eleven clinical cycles were performed , resulting in the birth of an unaffected girl . For one of the couples undergoing P52209 for P35240 , a spontaneous pregnancy ensued after five unsuccessful P52209 cycles . The couple underwent chorionic villus sampling ( CVS ) and the application of the same protocol as used during P52209 showed an unaffected fetus . CONCLUSION : In this work , we present the development and clinical application of P52209 for three cancer predisposition syndromes . Enhancement of retroviral infection in vitro by anti-Le(y) IgG : reversal by humanization of monoclonal mouse antibody . Monoclonal mouse IgG3 antibody ( P00519 364 ) against the carbohydrate Le(y) antigen enhanced infection in vitro with HTLV-1 and with HIV-1 when propagated in both transformed and normal lymphocytes . Enhancement was independent of complement , occurred with both lymphocytes and monocytes as target cells , and did not use either L(ey) epitopes on target cells for cross-linkage of virus to the cell or the Fc part of the antibody as a ligand for any cellular receptor . For enhancement to occur , binding of anti-Le(y) antibody to virus was required to take place before virus binding to its specific receptor with no indication of any alternative pathway of infection , as evidenced by abrogation of enhancement by anti- P01730 MAb or soluble recombinant P01730 , and also the inability of anti-Le(y) MAb to mediate HIV infection of DB01040 -2 cells in which HTLV-1/HIV pseudovirus infection was enhanced . While F(ab)2 fragments of P00519 364 also enhanced infection , a human/mouse chimeric antibody and a fully humanized antibody had no enhancing effect on free virus infection . We suggest that binding of anti-Le(y) P00519 364 or its F(ab)2 fragment induced a conformational change in the gp120 oligomers facilitating the process of infection , and that this function was abrogated by the IgG1 Fc of the chimeric and the humanized antibodies . The observations indicate that the non-paratope domains of antiviral antibodies can influence their function as neutralizing or enhancing for infection . [ New therapeutic strategy for autoimmune and chronic inflammatory disease based on clinical results using P05231 blocking therapy with a humanized anti- P05231 receptor antibody ] . Remarkable clinical effects were observed by P05231 blockage with a humanized anti P05231 receptor antibody in patients with Castleman 's disease , rheumatoid arthritis , and juvenile inflammatory arthritis . This evidence suggests that the hyper-function of P05231 is an essential key cytokine in the pathogenesis of the above diseases , in which many cytokines , chemokines , and inflammatory molecules are activated . We found , for example , P01375 blocking therapy showed a reduction of acute phase proteins , such as CRP and P0DJI8 , however , the P05231 blockade induced not only reduction but also normalization of CRP and P0DJI8 serum levels . To elucidate this in vivo phenomenon , we analyzed the expression of cytokine inducing CRP and P0DJI8 mRNA with the intracellular signal transduction mechanism in vitro . The results , indicated that the P05231 signal was essential though the activation of P40763 for the induction and augmentation of CRP or P0DJI8 by the associated stimulation with P01375 or IL-1 . Recently , it is now known that P05231 is a regulatory molecule in the induction of Th17 cells with TGF-beta . Therefore , P05231 blockage may potentially improve autoimmune diseases , beginning with the pathogenic initiation phase . We believe that unknown pathogenic inflammatory phenomena can be clarified using this analytical strategy and cytokine blocking therapy . Furthermore , in the future we hope to induce complete remission of autoimmune diseases by using cytokine blockage freely . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Elevated serum levels of soluble P78423 in patients with microscopic polyangiitis . OBJECTIVES : To test the hypothesis that P78423 contributes to the pathogenesis of microscopic polyangiitis . METHODS : Serum samples from 18 patients with microscopic polyangiitis ( DB00603 ) , who fulfilled the revised criteria of the American College of Rheumatology ( P10323 ) , were collected during both the newly diagnosed , untreated active disease states and inactive disease states . Also serum was from patients with large vessel vasculitis ( LVV ) , including giant cell arteritis ( n=4 ) and Takayasu arteritis ( n=3 ) , and from 52 healthy individuals . Soluble (s) P78423 levels in serum were measured using an enzyme-linked immunosorbent assay . Disease activity was assessed using Birmingham vasculitis activity scores ( BVAS ) . Expression of P49238 was examined by flow cytometry . RESULTS : Serum sCX3CL1 levels were significantly higher in DB00603 patients than in either LVV group or healthy individuals . The elevated sCX3CL1 levels seen in DB00603 patients correlated positively with BVAS , as well as with CRP levels and P03372 , and similarly increased expression of cell-surface P49238 was seen on peripheral blood P01730 and CD8 T cells from patients with DB00603 . Notably , sCX3CL1 levels and P49238 expression were diminished during clinical remission following treatment . CONCLUSION : Our findings suggest that P78423 may be involved in the pathogenesis of DB00603 , and may serve as a useful serologic marker of disease activity in systemic vasculitis . Mapping of 13 horse genes by fluorescence in-situ hybridization ( Q5TCZ1 ) and somatic cell hybrid analysis . We report fluorescence in-situ hybridization ( Q5TCZ1 ) and somatic cell hybrid mapping data for 13 different horse genes ( P01160 , P06729 , P10909 , P54108 , P05093 , P02679 , P18510 , P22301 , P45452 , PRM1 , P35354 , P01375 and P04637 ) . Primers for PCR amplification of intronic or untranslated regions were designed from horse-specific DNA or mRNA sequences in GenBank . Two different horse bacterial artificial chromosome ( BAC ) libraries were screened with PCR for clones containing these 13 Type I loci , nine of which were found in the libraries . BAC clones were used as probes in dual colour Q5TCZ1 to confirm their precise chromosomal origin . The remaining four genes were mapped in a somatic cell hybrid panel . All chromosomal assignments except one were in agreement with human-horse ZOO- Q5TCZ1 data and revealed new and more detailed information on the equine comparative map . P10909 was mapped by synteny to ECA2 while human-horse ZOO- Q5TCZ1 data predicted that P10909 would be located on ECA9 . The assignment of P18510 permitted analysis of gene order conservation between HSA2 and ECA15 , which identified that an event of inversion had occurred during the evolution of these two homologous chromosomes . Use of the ImmuKnow assay to evaluate the effect of alemtuzumab-depleting induction therapy on cell-mediated immune function after renal transplantation . BACKGROUND : Good outcomes after renal transplantation are dependent on effective immunosuppression while minimizing infection . DB00087 ( Campath or Campath-1H ) is an anti- P31358 humanized monoclonal IgG1 antibody which induces rapid and sustained depletion of circulating lymphocytes and has been effectively used as an immunosuppressant in post-transplant induction therapy . METHODS : We used the ImmuKnow assay to compare cell-mediated immune function in renal transplant patients treated with alemtuzumab or with conventional immunosuppressive tri-therapy . The ImmuKnow method determines the levels of adenosine triphosphate ( DB00171 ) released from P01730 cells following stimulation with a mitogen . RESULTS : We showed a statistically significant difference in the distribution of outcome after transplantation between the conventional and the Campath groups ( P = 0.010 ) . A significantly higher number of patients treated with alemtuzumab induction therapy were stable after transplantation compared to those treated with conventional immunosuppressive tri-therapy ( 96.6 vs. 75.7 % ) . DB00171 values were significantly higher in the conventional group compared to the Campath group at 180 days after transplantation ( P < 0.001 ) . DB00171 levels did not change significantly over time in clinically stable kidney recipients treated with alemtuzumab induction therapy ( P = 0.554 ) . CONCLUSIONS : The ImmuKnow assay is a useful tool for evaluating the global immune response in alemtuzumab-treated renal transplant patients . DB00087 -depleting induction therapy remains effective for at least 180 days . Long-term follow-up of symptomatic patients with lymphoplasmacytic lymphoma/Waldenström macroglobulinemia treated with the anti- P31358 monoclonal antibody alemtuzumab . P31358 is expressed on malignant cells in lymphoplasmacytic lymphoma ( P06858 ) , including IgM-secreting Waldenström macroglobulinemia ( WM ) . We examined the activity of alemtuzumab in 28 symptomatic P06858 ( 27 IgM and 1 IgA ) patients . The median prior number of therapies for these patients was 2 ( range , 0-5 ) and 43 % had refractory disease . Patients received alemtuzumab at 30 mg IV 3 times weekly for up to 12 weeks after test dosing , and also received hydrocortisone , acyclovir , and Bactrim or equivalent prophylaxis . Patients had a complete response ( n = 1 ) , a partial response ( n = 9 ) , or a MR ( n = 11 ) for an overall and major response rate of 75 % and 36 % , respectively . Median serum Ig decreased from 3510 to 1460 mg/dL ( P < .001 at best response ) . With a median follow-up of 64 months , the median time to progression was 14.5 months . Hematologic and infectious complications , including CMV reactivation , were more common in previously treated patients and were indirectly associated with 3 deaths . Long-term follow-up revealed late-onset autoimmune thrombocytopenia ( AITP ) in 4 patients at a median of 13.6 months after therapy , which contributed to 1 death . DB00087 is an active therapy in patients with P06858 , but short- and long-term toxicities need to be carefully weighed against other available treatment options . Late AITP is a newly recognized complication of alemtuzumab in this patient population . This study is registered at www.clinicaltrials.gov as NCT00142181 . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . Fatal rhabdomyolysis in a patient with liver cirrhosis after switching from simvastatin to fluvastatin . P04035 inhibitors ( statins ) are widely used to treat hypercholesterolemia . Among the adverse effects associated with these drugs are statin-associated myopathies , ranging from asymptomatic elevation of serum creatine kinase to fatal rhabdomyolysis . DB01095 -induced fatal rhabdomyolysis has not been previously reported . We describe here a patient with liver cirrhosis who experienced fluvastatin-induced fatal rhabdomyolysis . This patient had been treated with simvastatin ( 20 mg/day ) for coronary artery disease and was switched to fluvastatin ( 20 mg/day ) 10 days before admission . He was also taking aspirin , betaxolol , candesartan , lactulose , and entecavir . Rhabdomyolysis was complicated and continued to progress . He was treated with massive hydration , urine alkalization , intravenous furosemide , and continuous renal replacement therapy for acute renal failure , but eventually died due to rhabdomyolysis complicated by hepatic failure . In conclusion , fluvastatin should be used with caution in patients with liver cirrhosis , especially with other medications metabolized with P11712 . DB00087 induction of intracellular signaling and apoptosis in malignant B lymphocytes . The molecular changes induced by alemtuzumab following binding of P31358 on B tumor cells were investigated . DB00087 alone had no detectable impact on cell signaling but cross-linking of alemtuzumab on the surface of B tumor lines with anti-human Fc antibodies induced a transient Ca(2+) flux followed by phosphorylation of several kinases involved in stress and survival pathways , and expression of associated proteins including P01375 -α . Cross-linking of alemtuzumab also induced capping and caspase-dependent apoptosis of the tumor lines . When using primary cells from B-CLL patients , alemtuzumab alone was capable of inducing protein phosphorylation and apoptosis through the cross-linking of alemtuzumab by FcγRIIb receptors on B-CLL cells . Apoptosis was prevented by blocking of FcγRIIb receptors with anti-CD32 antibody . Overall , our results indicate that cross-linking of alemtuzumab on B tumor cells can occur naturally through Fc receptor interaction and leads to the activation of specific cellular pathways and induction of apoptosis . Informative gene network for chemotherapy-induced peripheral neuropathy . BACKGROUND : Host genetic variability has been implicated in chemotherapy-induced peripheral neuropathy ( CIPN ) . A dose-limiting toxicity for chemotherapy agents , CIPN is also a debilitating condition that may progress to chronic neuropathic pain . We utilized a bioinformatics approach , which captures the complexity of intracellular and intercellular interactions , to identify genes for CIPN . METHODS : Using genes pooled from the literature as a starting point , we used Ingenuity Pathway Analysis ( IPA ) to generate gene networks for CIPN . RESULTS : We performed IPA core analysis for genes associated with platinum- , taxane- and platinum-taxane-induced neuropathy . We found that P05231 , P01375 , P10145 , P01584 and P27361 /2 were the top genes in terms of the number of connections in platinum-induced neuropathy and P04637 , MYC , P09874 , O75791 MAPK and P01375 for combined taxane-platinum-induced neuropathy . CONCLUSION : Neurotoxicity is common in cancer patients treated with platinum compounds and anti-microtubule agents and CIPN is one of the debilitating sequela . The bioinformatic approach helped identify genes associated with CIPN in cancer patients . Extrapulmonary small cell : a novel case of small cell carcinoma of the thyroid gland . Neuroendocrine tumors comprise a large group of malignancies which share unique morphological features and are characterized by the presence of neuroendocrine markers such as synaptophysin , chromogranin-A , and CD56 ( N- P62158 ) , ranging from indolent tumors , such as carcinoid tumors , to aggressive tumors , such as small cell carcinoma . The lung is the most common site for primary neuroendocrine tumors . Extrapulmonary primary sites of small cell carcinoma are rare but have been documented arising from various sites including esophagus , stomach , colon and rectum , gallbladder , thymus , salivary gland , ovary , cervix , bladder , prostate , and skin . We present a case of small cell carcinoma arising from the thyroid gland , a site not previously described in the literature . A 59-year-old woman presented with a thyroid mass , which , after resection , showed small cell morphology and positive immunostains for Q15669 -1 , synaptophysin , chromogranin-A , CD56 , etc . Five months after diagnosis , she had widely metastatic disease . After a near-complete response to the first chemo-treatment , her disease progressed . Following local radiation and more rounds of chemotherapy , she succumbed to the disease , 15 months after diagnosis . Our patient had no pulmonary lesions at the time of diagnosis to suggest metastasis from the lung . Much like its pulmonary counterparts , this small cell carcinoma of primary thyroid origin displayed an aggressive clinical course and poor outcome . Although it shows early sensitivity to chemotherapy , small cell carcinoma remains a difficult-to-treat cancer with a poor prognosis and can rarely be seen originating in organs outside of the lung . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . t(1;3)(p36; P38936 ) is a recurring therapy-related translocation . Chromosome bands 1p36 and 3p21 are known to be recurring breakpoints in therapy-related ( t- ) leukemia . We identified a recurring translocation , t(1;3)(p36; P38936 ) , in eight patients with various hematologic malignancies : three patients with ALL , one with chronic myelogenous leukemia ( CML ) in accelerated phase ( AP ) , two with P43034 , and two with AML(M3) . Five of the eight patients had a history of chemotherapy , including alkylating agents in three , before the translocation was detected . In two of these five patients , the t(1;3)(p36; P38936 ) emerged only at relapse or in the accelerated phase of CML . The karyotypes of the patients were complex , including -7 and structural abnormalities of 5q , 6q , 7q , 9p , and 11q23 . Survival time varied among patients ( 25 days to more than 16 years ) . Using Q5TCZ1 with 13 1p35-36 cosmid probes ( tel-FB12- P35218 -G7-FD2- P21554 -ED8-FD9-G32- P48751 -G50-AD8-GG4-G43-cen ) , we delineated the 1p36 breakpoint in two patients with P43034 and ALL as lying between FB12 and FD2 ( between BAC47P3 and PAC963K15 ) , with a small deletion near the breakpoint in both cases . In the patient with P43034 , there was also a deletion at 3p21.3 , as detected with the cosmid probe cosNRL9 . The results of the present study suggest that t(1;3)(p36; P38936 ) in hematologic diseases is associated with prior exposure to mutagens , including alkylating agents . Involvement of neutrophils and natural killer cells in the anti-tumor activity of alemtuzumab in xenograft tumor models . DB00087 is a recombinant humanized IgG1 monoclonal antibody directed against P31358 , an antigen expressed on the surface of normal and malignant B and T lymphocytes . DB00087 is approved for the treatment of B-cell chronic lymphocytic leukemia ( B-CLL ) , but the exact mechanism by which the antibody depletes malignant lymphocytes in vivo is not clearly defined . To address this issue , the anti-tumor activity of alemtuzumab was studied in disseminated and subcutaneous xenograft tumor models . The density of P31358 target antigen on the surface of tumor cells appeared to correlate with the anti-tumor activity of alemtuzumab . Deglycosylation of alemtuzumab resulted in a loss of cytotoxicity in vitro and was found to abolish anti-tumor activity in vivo . Individual inactivation of effector mechanisms in tumor-bearing mice indicated that the protective activity of alemtuzumab in vivo was primarily dependent on ADCC mediated by neutrophils and to a lesser extent NK cells . Increasing the number of circulating neutrophils by treatment with G- P04141 enhanced the anti-tumor activity of the antibody , thus providing further evidence for the involvement of neutrophils as effector cells in the activity of alemtuzumab . DB00087 in peripheral T-cell malignancies . The humanized monoclonal antibody CAMPATH-1H ( alemtuzumab ) binds to the P31358 antigen , a glycoprotein that is widely expressed on normal and malignant B- and T-lymphocytes . Over the past 5 years , a number of trials have demonstrated that alemtuzumab has clinical activity in mature T-cell diseases such as T-cell prolymphocytic leukemia ( T-PLL ) and cutaneous T-cell lymphoma ( CTCL ) . In heavily pretreated relapsed/refractory patients alemtuzumab induced responses in more than two thirds of T-PLL and more than 50 % of CTCL patients . Responding patients had improved survival compared to nonresponders . DB00087 is particularly effective in clearing malignant lymphocytes from peripheral blood and bone marrow and may therefore facilitate stem-cell transplantation ( P09683 ) in selected patients . The toxicity profile for the antibody is acceptable ; the major complications are infusional reactions , which generally subside after the first 1-2 weeks of therapy , and prolonged lymphopenia associated with reactivation of viruses . These can be minimized by careful monitoring and the use of prophylactic therapy . Future studies will be directed toward : alternative routes ( subcutaneous ) and schedules of administration ; use as first-line therapy ; combination strategies with conventional chemotherapy ; and use of alemtuzumab to purge minimal residual bone-marrow disease prior to P09683 . Serotonin skews human macrophage polarization through P41595 and P34969 . Besides its role as a neurotransmitter , serotonin ( 5-hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT(1-7) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 production , upregulated the expression of M2 polarization-associated genes ( P05120 , P07996 , Q9NY15 , Q86Y22 ) , and reduced the expression of M1-associated genes ( P08476 , P41597 , P39900 , P05121 , P29016 , O94788 ) . Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2( P09603 ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7) , whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies . ER stress is associated with dedifferentiation and an epithelial-to-mesenchymal transition-like phenotype in PC Cl3 thyroid cells . Conditions perturbing the homeostasis of the endoplasmic reticulum ( ER ) cause accumulation of unfolded proteins and trigger ER stress . In PC Cl3 thyroid cells , thapsigargin and tunicamycin interfered with the folding of thyroglobulin , causing accumulation of this very large secretory glycoprotein in the ER . Consequently , mRNAs encoding P11021 and P17861 were induced and spliced , respectively . In the absence of apoptosis , differentiation of PC Cl3 cells was inhibited . mRNA and protein levels of the thyroid-specific genes encoding thyroglobulin , thyroperoxidase and the sodium/iodide symporter and of the genes encoding the thyroid transcription factors Q15669 -1 , O00358 and Pax-8 were dramatically downregulated . These effects were , at least in part , transcriptional . Moreover , they were selective and temporally distinct from the general and transient Q9NZJ5 -dependent translational inhibition . Thyroid dedifferentiation was accompanied by changes in the organization of the polarized epithelial monolayer . Downregulation of the mRNA encoding P12830 , and upregulation of the mRNAs encoding vimentin , alpha-smooth muscle actin , alpha(1)(I) collagen and O95863 /SIP1 , together with formation of actin stress fibers and loss of trans-epithelial resistance were found , confirming an epithelial-mesenchymal transition ( EMT ) . The thyroid-specific and epithelial dedifferentiation by thapsigargin or tunicamycin were completely prevented by the Q99463 inhibitor of Src-family kinases and by stable expression of a dominant-negative Src . Together , these data indicate that ER stress induces dedifferentiation and an EMT-like phenotype in thyroid cells through a Src-mediated signaling pathway . Resistance to diet-induced adiposity in cannabinoid receptor-1 deficient mice is not due to impaired adipocyte function . BACKGROUND : Overactivity and/or dysregulation of the endocannabinoid system ( ECS ) contribute to development of obesity . In vitro studies indicate a regulatory role for the cannabinoid receptor 1 ( P21554 ) in adipocyte function and P21554 -receptor deficient ( P21554 -/- ) mice are resistant to high fat diet-induced obesity . Whether this phenotype of P21554 -/- mice is related to altered fat metabolism in adipose tissue is unknown . METHODS : We evaluated adipose tissue differentiation/proliferation markers and quantified lipogenic and lipolytic activities in fat tissues of P21554 -/- and P21554 +/+ mice fed a high-fat ( HF ) or a high-fat/fish oil ( HF/FO ) diet as compared to animals receiving a low-fat chow diet . Comparison between HF diet and HF/FO diet allowed to investigate the influence of dietary fat quality on adipose tissue biology in relation to P21554 functioning . RESULTS : The adiposity-resistant phenotype of the P21554 -/- mice was characterized by reduced fat mass and adipocyte size in HF and HF/FO-fed P21554 -/- mice in parallel to a significant increase in energy expenditure as compared to P21554 +/+ mice . The expression levels of adipocyte differentiation and proliferation markers were however maintained in these animals . Consistent with unaltered lipogenic gene expression , the fatty acid synthesis rates in adipose tissues from P21554 -/- and P21554 +/+ mice were unchanged . Whole-body and adipose-specific lipoprotein lipase ( P06858 ) activities were also not altered in P21554 -/- mice . CONCLUSIONS : These findings indicate that protection against diet-induced adiposity in P21554 -deficient mice is not related to changes in adipocyte function per se , but rather results from increased energy dissipation by oxidative and non-oxidative pathways . DB00087 ( Campath-1H ) induction therapy and dendritic cells : Impact on peripheral dendritic cell repertoire in renal allograft recipients . Dendritic cells ( DC ) are the most potent antigen-presenting cells ( P25054 ) and are pivotal for initiating allograft immunity . Recently , particular DC subsets have been implicated also in allogeneic T cell hyporesponsiveness . DB00087 ( anti- P31358 , Campath-1H ) is a novel T cell depleting antibody that is currently under investigation for the use in allogeneic organ transplantation . While recent studies demonstrated a conspicuous effect of alemtuzumab on peripheral DC in clinical graft-versus-host disease , its efficiency in patients receiving allogeneic organ transplants is still undefined . In the present study we assessed the peripheral DC repertoire in kidney transplant recipients after either alemtuzumab induction therapy followed by FK506 monotherapy or after conventional immunosuppression ( FK506 , mycophenolate mofetil and steroids ) without any induction agent . Induction with alemtuzumab caused a strong and sustained reduction of the total number of peripheral DC and a significant shift from myeloid to plasmacytoid DC subsets ( mDC/pDC ratio ) as early as 1 month post-transplantation . These data show that alemtuzumab induction targets the peripheral DC repertoire , which might add another mechanism allowing immunosuppressive drug minimization . Further studies are warranted to further elucidate the functional significance of these finding in the setting of allogeneic organ transplantation . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 , CD8 and forkhead box P09131 ( Foxp3 ) , respectively . P14136 , Iba1 and P34810 -immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed . DB00087 as a bridge to allogeneic P09683 in atypical hemophagocytic lymphohistiocytosis . BACKGROUND : A 39-year-old woman with no relevant medical or family history was admitted to hospital with episodic fever , which persisted despite antibiotic therapy . Other notable findings at admission were splenomegaly , pancytopenia , hyponatremia , elevated levels of liver enzymes , hyperferritinemia and hypofibrinogenemia . INVESTIGATIONS : Physical examination , laboratory tests , rheumatic marker serology , pathogen detection assays , complete blood counts , measurement of levels of ferritin , fibrinogen , triglycerides and soluble CD25 , natural killer cell functional studies , P14222 mutation analysis , renal biopsy , bone marrow biopsy , CT imaging of the chest and abdomen . DIAGNOSIS : Idiopathic , atypical hemophagocytic lymphohistiocytosis . MANAGEMENT : Initial treatment with antibiotics was followed by immunosuppressive therapy ( including intravenous immunoglobulin , ciclosporin , infliximab , corticosteroids and etoposide ) . Remission was achieved by treatment with the anti- P31358 monoclonal antibody , alemtuzumab , after which allogeneic stem-cell transplantation ( with reduced-intensity conditioning treatment and graft-versus-host disease prophylaxis ) resulted in a definitive cure . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Ex vivo-expanded cynomolgus macaque regulatory T cells are resistant to alemtuzumab-mediated cytotoxicity . DB00087 ( Campath-1H ) is a humanized monoclonal antibody ( Ab ) directed against P31358 that depletes lymphocytes and other leukocytes , mainly by complement-dependent mechanisms . We investigated the influence of alemtuzumab ( i ) on ex vivo-expanded cynomolgus monkey regulatory T cells ( Treg ) generated for prospective use in adoptive cell therapy and ( ii ) on naturally occurring Treg following alemtuzumab infusion . Treg were isolated from PBMC and lymph nodes and expanded for two rounds . P31358 expression , binding of alemtuzumab and both complement-mediated killing and Ab-dependent cell-mediated cytotoxicity ( ADCC ) were compared between freshly isolated and expanded Treg and effector T cells . Monkeys undergoing allogeneic heart transplantation given alemtuzumab were monitored for Treg and serum alemtuzumab activity . Ex vivo-expanded Treg showed progressive downregulation of P31358 expression , absence of alemtuzumab binding , minimal change in complement inhibitory protein ( P15529 ) expression and no complement-dependent killing or ADCC . Infusion of alemtuzumab caused potent depletion of all lymphocytes , but a transient increase in the incidence of circulating Treg . After infusion of alemtuzumab , monkey serum killed fresh PBMC , but not expanded Treg . Thus , expanded cynomolgus monkey Treg are resistant to alemtuzumab-mediated , complement-dependent cytotoxicity . Furthermore , our data suggest that these expanded monkey Treg can be infused into graft recipients given alemtuzumab without risk of complement-mediated killing . Statin decreases endothelial microparticle release from human coronary artery endothelial cells : implication for the Rho-kinase pathway . OBJECTIVE : Elevated plasma levels of endothelial microparticles ( EMPs ) are associated with the presence of clinical atherosclerosis . Considering the anti-inflammatory properties of P04035 inhibitors on the endothelium , we studied the effect of fluvastatin on the release of EMPs in cultured human coronary artery endothelial cells ( HCAEC ) . METHODS AND RESULTS : EMPs were generated in P01375 -activated HCAECs . The absolute number of EMPs was enumerated using a novel two-color flow cytometric immunostaining technique with TruCount beads as an internal reference . EMPs are defined as EC membrane vesicles ( 1-2 microm in size ) with a characteristic immunophenotype . The addition of fluvastatin to P01375 -activated HCAECs significantly suppressed Q7L5Y9 release . DB01095 suppressed P01375 -induced Rho activation . The Rho-kinase inhibitor , Y-27632 , reproduced the effect of statin . CONCLUSION : Q7L5Y9 release from P01375 -activated HCAECs is suppressed by fluvastatin . In addition , the Rho/Rho-kinase may play an important role in modulating Q7L5Y9 release . Real life experience with alemtuzumab treatment of patients with lower-risk P43034 and a hypocellular bone marrow . Immunosuppressive therapy is a therapeutic option for selected low-risk myelodysplastic syndromes ( P43034 ) patients . Besides standard treatment protocols that include ATG and Q13216 , the humanized P31358 antibody alemtuzumab has been shown to have efficacy in P43034 treatment . We report our experience with alemtuzumab in nine P43034 RCMD patients . All patients had a hypocellular bone marrow with a blast count < 5 % and were classified as intermediate-1 according to the IPSS . We found a response in five patients ( 60 % ) ; three patients achieved a complete remission 3 and 6 months after the treatment with alemtuzumab , and two patients showed a haematological improvement . DB00087 was administered in a 10-mg dosage for 10 days . Treatment was well tolerated , and no severe side effects were observed . We could confirm the finding that the alemtuzumab is effective and save selected P43034 patients . Due to the promising results , further studies , especially with regard to long-term survival and risk of leucemic progression should be initiated . DB00087 ( anti- P31358 monoclonal antibody ) as single-agent therapy in patients with relapsed/refractory chronic lymphocytic leukaemia ( CLL ) -a single region experience on consecutive patients . DB00087 , a humanized anti- P31358 monoclonal antibody , is used in patients with refractory chronic lymphocytic leukaemia ( CLL ) . We report results in health care with alemtuzumab on consecutive , advanced-stage patients from a well-defined geographical region . Records from 1,301 patients ( Stockholm-Cancer-Registry 1991-2010 ) identified 56 relapsed/refractory patients treated with alemtuzumab . Median age was 69 years , 88 % had advanced Rai-stage with median 3 prior therapies . One fourth had bulky lymphadenopathy and 73 % were refractory to purine analogues . Median treatment length was 11.6 weeks . Median cumulative dose was 930 mg , significantly higher ( p = 0.0277 ) for responders . Overall response-rate ( ORR ) was 43 % ; 32.5 % , 50 % and 87.5 % in the Refractory , Purine analogue relapsed and Relapsed/Other subgroup , respectively . Response rate was significantly associated with subgroup ( p = 0.0104 ) . Good performance status ( PS ) was associated with better response rate ( p = 0.0227 ) . Median time-to-treatment-failure ( Q15669 ) ( months ) was 7.8 months , significantly ( p < 0.0001 ) longer for responders ( 13.4 ) Major infections occurred in 36 % . Median overall survival was 22.5 months ( range 0.4-74.3 ) . Positive predictive factors were good PS ( p < 0.0001 ) and fewer previous therapies ( p = 0.0038 ) . Twenty percent were retreated with alemtuzumab with an ORR of 54.5 % , and a Q15669 of 7.1 months . A high cumulative dose/longer duration of therapy and a relatively high response rate was observed compared to previous reports . Optimal patient identification and management may result in avoidance of early discontinuation and possibly better outcomes . Circulating endothelial progenitor cells decreased in patients with sclerodermatous chronic graft-versus-host disease . Chronic graft-versus-host disease ( cGVHD ) is a common late complication of allogeneic stem cell transplantation ( allo- P09683 ) . Some cGVHD patients develop skin lesions , and the skin lesions in sclerodermatous cGVHD ( s-cGVHD ) patients resemble those in progressive systemic sclerosis ( PSS ) , which is characterized by impaired production of circulating endothelial progenitor cells ( EPCs ) . We investigated , retrospectively , whether low EPC production may promote the development of sclerodermatous lesions in cGVHD . Peripheral blood ( PB ) was obtained from 14 healthy volunteers and 27 allo- P09683 patients . Five patients developed s-cGVHD . P28906 (+) cells were purified by using the magnetic cell-sorting separation system , and the P28906 (+)/CD133(+)/vascular endothelial growth factor ( P15692 ) receptor-2(+) EPCs were quantified . The endothelial cell colony-formation potential was evaluated . Serum P15692 and basic fibroblast growth factor ( b-FGF ) concentrations were measured by ELISA . The s-cGVHD patients had significantly lower median circulating EPCs frequencies than non-s-cGVHD patients or control ( 145 of 20 mL [ interquartial range-IQR 107-193 ] versus 1083.5 [ IQR 669.3-2151 ] ; P = .0023 , and versus 1530.5 [ IQR 961.3-2158 ] ; P = .0012 , respectively ) . They also had impaired median endothelial-forming ability compared to non-s-cGVHD patients or controls ( 3.8 [ IQR 1.0-4.3 ] versus 12.8 [ IQR 8.8-28.8 ] , and versus 26.4 [ IQR 23.6-30.6 ] , respectively ; P = .0012 ) . Their P15692 and b-FGF serum levels were also higher than in controls . In conclusion , s-cGVHD patients show findings consistent with those seen in PSS with impaired vasculogenesis that may limit blood perfusion and may contribute to the development of sclerodermatous lesions . A mechanistic rationale for combining alemtuzumab and rituximab in the treatment of ALL . B-lineage acute lymphoblastic leukemia ( ALL ) may express P31358 and P11836 . DB00087 ( ALM ) and rituximab ( RTX ) are therapeutic antibodies directed against P31358 and P11836 , respectively , but showed limited activity against ALL in clinical trials . The mechanisms for the impaired responses remained unclear . We studied expression of P31358 and P11836 on ALL cells and found that most cases coexpressed P31358 and P11836 . However , distinct P31358 -negative ( P31358 (-) ) subpopulations were detected in most cases as the result of defective glycophosphatidyl-inositol anchoring . Although ALM efficiently eradicated P31358 -positive ( P31358 (+) ) cells in NOD/scid mice engrafted with primary human ALL , P31358 (-) subclones escaped therapy . In the same model , RTX showed limited activity resulting from occurrence of P11836 down-modulation . However , P31358 (-) cells concurrently lacked the glycophosphatidyl-inositol-anchored complement regulators P08174 and P13987 and showed increased susceptibility to RTX-mediated complement-dependent cytotoxicity in vitro . At the same time , ALM was shown to inhibit down-modulation of P11836 in response to RTX by depleting the trogocytic capacity of phagocytic cells . Probably because of these complementary mechanisms , combined administration of ALM and RTX induced complete responses in vivo . Based on these data , we propose a mechanistic rationale for combined application of RTX and ALM in ALL . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Antibody-based therapy of non-Hodgkin 's lymphoma . Monoclonal antibodies ( mAb ) have dramatically advanced our ability to treat non-Hodgkin 's lymphoma ( Q9NZ71 ) , and there has been a virtual explosion of clinical data regarding their use . DB00073 is a humanized anti- P11836 mAb and has significant single agent activity in follicular lymphoma , and to a lesser extent in mantle-cell and diffuse large B-cell lymphoma ( DLCL ) . DB00073 appears to have synergistic activity with cytotoxic chemotherapy and the combination has recently demonstrated improved rates of complete remission ( CR ) and overall survival in older patients with DLCL . DB00087 ( Campath-1H ) is a humanized mAb targeting P31358 and has recently been approved in the USA for the treatment of fludarabine-refractory B-cell chronic lymphocytic leukaemia . Impressive activity has also been demonstrated in T-cell prolymphocytic leukaemia and mycosis fungoides . The radioconjugated anti- P11836 mAbs ibritumomab tiuxetan and I131-tositumomab also have impressive clinical activity in low-grade B-cell Q9NZ71 , and the former has demonstrated superior CR rates to rituximab . Myelosuppression is more significant however , and their place in the treatment algorithm remains to be clearly defined . Other immunotoxins ( e.g. BL22 ) and mAb against alternate targets ( e.g. epratuzumab , humanized anti- P20273 ) are in development . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation .

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Modulation of sympathetic nerve activity by microinjection of the P08908 receptor agonist 8-OH-DPAT into the rostroventrolateral medulla . In the present study , renal sympathetic nerve activity was recorded simultaneously with sympathetic nerve activity to skeletal muscle vasculature to determine if the sympatho-inhibition evoked by microinjection of the P08908 receptor agonist 8-hydroxy-2-(di-n-propylamino)teralin ( 8-OH-DPAT ) into the rostroventrolateral medulla ( RVLM ) was uniform or regional . Three patterns of sympatho-inhibition were observed in these sympathetic outflows and the type of response depended upon location of microinjection within the subretrofacial nucleus ( P11831 ) . Inhibition of renal nerve activity only was elicited by microinjections at rostral sites at the caudal pole of the facial nucleus . In contrast , inhibition of muscle sympathetic nerve activity was evoked from more caudal injections at the rostral pole of the inferior olives . Microinjection in the area between these two regions produced inhibition of both sympathetic outflows . This study demonstrates that differential inhibition of regional sympathetic outflows can be elicited by microinjection of the P08908 receptor agonist 8-OH-DPAT into the RVLM . These data suggests that this modulation is due to differences in anatomical arrangement of the medullary neurons rather than differences in neuron sensitivity to the serotonergic agonist . Loss of the candidate tumor suppressor Q14201 triggers acute cellular senescence via the P29323 - O15054 -p16(INK4a) signaling axis . The B-cell translocation gene 3 ( Q14201 ) is a member of the antiproliferative BTG gene family and a downstream target of p53 . Q14201 also binds and inhibits Q01094 . Although it connects functionally two major growth-regulatory pathways , the physiological role of Q14201 remains largely uncharacterized . Here , we present evidence that loss of Q14201 in normal cells induced cellular senescence , which was correlated with enhanced P29323 - P05412 signaling and elevated expression of the histone H3K27me3 demethylase O15054 / O15054 , leading to acute induction of p16(INK4a) . Importantly , we also found that Q14201 expression is specifically downregulated in prostate cancer , thus providing a physiological link with human cancers . Our data suggest that Q14201 may have a fail-safe role against tumorigenic progression . Vascular changes after cardiac surgery : role of NOS , P36551 , kinases , and growth factors . Cardiovascular disease remains the leading cause of mortality in the industrialized world . Despite advances in pharmacotherapy and catheter based interventions , coronary artery bypass grafting remains an essential therapeutic modality . The majority of coronary artery bypass operations , as well as other cardiac surgical procedures require the use of ischemic cardioplegic arrest and cardiopulmonary bypass , both of which result in iatrogenic injury to the vasculature and microcirculation . This injury can manifest as impaired vasorelaxation or vasoconstriction , depending upon the organ system involved , resulting in impaired tissue perfusion and the development of edema . Key to this dysfunction are changes in the following : nitric oxide signaling secondary to changes in P29474 and P35228 expression and activity , cyclooxygenase function with increases in pro-inflammatory P35354 activity , alterations in Protein Kinase C and Mitogen Activated Protein Kinase signaling , and an increase in Vascular Endothelial Growth Factor expression increasing vascular permeability and dilatation . This review discusses our current understanding of cardioplegia and cardiopulmonary bypass induced changes in the vasculature , and therapeutic interventions aimed at modulating the altered signaling pathways . Compound FLZ inhibits lipopolysaccharide-induced inflammatory effects via down-regulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways in RAW264.7 macrophages . AIM : The aim of this study was to investigate the effect of the squamosamide derivative FLZ ( N-2-(4-hydroxy-phenyl)-ethyl-2-(2,5-dimethoxy-phenyl)-3-(3-methoxy-4-hydroxy-phenyl)-acrylamide ) on lipopolysaccharide ( LPS ) -induced inflammatory mediator production and the underlying mechanism in RAW264.7 macrophages . METHODS : RAW264.7 cells were preincubated with non-toxic concentrations of compound FLZ ( 1 , 5 , and 10 micromol/L ) for 30 min and then stimulated with 10 microg/L LPS . The production of nitric oxide ( NO ) , the expression of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase 2 ( P35354 ) , and the activation of nuclear factor kappa-B ( NF-kappaB ) and mitogen-activated protein kinase ( MAPK ) pathways were examined . RESULTS : FLZ significantly inhibited the LPS-induced production of NO , as well as the expression of P35228 and P35354 at both the RNA and the protein levels in RAW264.7 cells . The LPS-induced increase in the DNA binding activity of NF-kappaB and activator protein 1 ( AP-1 ) , the nuclear translocation of NF-kappaB p65 , the degradation of the inhibitory kappaBalpha protein ( P25963 ) and the phosphorylation of P25963 , O15111 ( IKK ) alpha/beta , c-Jun NH(2)-terminal kinase ( JNK ) and p38 MAPKs were all suppressed by FLZ . However , the phosphorylation of extracellular signal-regulated kinase ( P29323 ) was not affected . Further study revealed that FLZ inhibited the phosphorylation of transforming growth factor-beta ( TGF-beta ) -activated kinase 1 ( TAK1 ) , which is an upstream signaling molecule required for IKKalpha/beta , JNK and p38 activation . CONCLUSION : FLZ inhibited the LPS-induced production of inflammatory mediators at least partly through the downregulation of the P50750 -IKK and P50750 -JNK/p38MAPK pathways . Pulmonary arterial dysfunction in insulin resistant obese Zucker rats . BACKGROUND : P01308 resistance and obesity are strongly associated with systemic cardiovascular diseases . Recent reports have also suggested a link between insulin resistance with pulmonary arterial hypertension . The aim of this study was to analyze pulmonary vascular function in the insulin resistant obese Zucker rat . METHODS : Large and small pulmonary arteries from obese Zucker rat and their lean counterparts were mounted for isometric tension recording . mRNA and protein expression was measured by RT-PCR or Western blot , respectively . KV currents were recorded in isolated pulmonary artery smooth muscle cells using the patch clamp technique . RESULTS : Right ventricular wall thickness was similar in obese and lean Zucker rats . Lung Q13873 , KV1.5 and 5- Q13049 receptor mRNA and protein expression and KV current density were also similar in the two rat strains . In conductance and resistance pulmonary arteries , the similar relaxant responses to acetylcholine and nitroprusside and unchanged lung P29474 expression revealed a preserved endothelial function . However , in resistance ( but not in conductance ) pulmonary arteries from obese rats a reduced response to several vasoconstrictor agents ( hypoxia , phenylephrine and 5-HT ) was observed . The hyporesponsiveness to vasoconstrictors was reversed by L-NAME and prevented by the P35228 inhibitor 1400W . CONCLUSIONS : In contrast to rat models of type 1 diabetes or other mice models of insulin resistance , the obese Zucker rats did not show any of the characteristic features of pulmonary hypertension but rather a reduced vasoconstrictor response which could be prevented by inhibition of P35228 . 17 DB00783 overcomes a P55008 block induced by P04035 inhibitors and fosters cell cycle progression without inducing P27361 and -2 Q96HU1 kinases activation . P04035 inhibitors , such as DB00227 and Simvastatin , cause cell cycle arrest by interfering with the mitogenic activity of mitogens present in culture media . Cells are induced to pause in P55008 and can readily resume growth upon removal of the enzymatic block . DB00286 , acting via their nuclear receptor , are mitogens for different normal and transformed cell types , where they foster cell cycle progression and cell division . In estrogen-responsive MCF-7 human breast cancer cells , but not in non responsive cells , 17 beta-estradiol ( E2 ) induces cells arrested with DB00227 or Simvastatin to proliferate in the presence of inhibitor , without restoring P04035 activity or affecting the protein prenylation pattern . Mitogenic stimulation of P55008 -arrested MCF-7 cells with E2 includes primary transcriptional activation of c-fos , accompanied by transient binding in vivo of the estrogen receptor and/or other factors to the ERE and the estrogen-responsive DNA region of this proto-oncogene , as detected by dimethylsulphate genomic footprinting analysis . Mitogenic stimulation of growth-arrested MCF-7 cells by E2 occurs , under these conditions , without evident activation of P27361 and -2 kinases , and thus independently from the mitogen-responsive signal transduction pathways that converge on these enzymes . Histone deacetylase inhibitors suppress the induction of c-Jun and its target genes including P35354 . P35354 ( P35354 ) is considered to be a target for anticancer therapy . Histone deacetylase ( HDAC ) inhibitors exhibit antitumor activity , but the mechanisms of action are incompletely understood . We investigated whether HDAC inhibitors blocked AP-1-mediated activation of P35354 transcription . Trichostatin A and suberoylanilide hydroxamic acid , two structurally related inhibitors of HDAC activity , blocked AP-1-mediated induction of P35354 expression and prostaglandin E2 biosynthesis . Chromatin immunoprecipitation assays indicated that HDAC inhibitors suppressed c-Jun binding to the P35354 promoter and thereby blocked transcription . The observed reduction in binding reflected reduced levels of c-Jun . HDAC inhibitors suppressed the induction of c-jun transcription by blocking the recruitment of the preinitiation complex ( RNA polymerase II and Q00403 ) to the c-jun promoter . O15379 but not Q13547 or Q92769 was required for AP-1-mediated stimulation of c-jun expression . Because HDAC inhibitors suppressed the induction of c-jun gene expression , resulting in reduced P35354 transcription , it was important to determine whether other known AP-1 target genes were also modulated . P12004 D1 and collagenase-1 are AP-1-dependent genes that have been implicated in carcinogenesis . HDAC inhibitors suppressed the induction of both cyclin D1 and collagenase-1 transcription by inhibiting the binding of c-Jun to the respective promoters . Taken together , these results suggest that HDAC inhibitors block the induction of c-jun transcription by inhibiting the recruitment of the preinitiation complex to the c-jun promoter . This led , in turn , to reduced expression of several activator protein-1-dependent genes ( P35354 , cyclin D1 , collagenase-1 ) . These findings provide new insights into the mechanisms underlying the antitumor activity of HDAC inhibitors . 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 . Effects of dutasteride on the expression of genes related to androgen metabolism and related pathway in human prostate cancer cell lines . Androgens play an important role in controlling the growth of the normal prostate gland and in the pathogenesis of benign prostate hyperplasia , and prostate cancer . Although testosterone is the main androgen secreted from the testes , dihydrotestosterone ( DB02901 ) , a more potent androgen converted from testosterone by 5alpha-reductase isozymes , type I and II , is the major androgen in the prostate cells . The aim of this study is to investigate the cellular and molecular effects of dutasteride , a potent inhibitor of 5alpha-reductase type I and type II , in androgen-responsive ( LNCaP ) and androgen-unresponsive ( DU145 ) human prostate cancer(PCa) cell lines . The expression pattern of 190 genes , selected on the basis of their proved or potential role in prostate cancerogenesis related to androgen signalling , were analysed using a low density home-made oligoarray ( AndroChip 2 ) . Our results show that dutasteride reduces cell viability and cell proliferation in both cell lines tested . AndroChip 2 gene signature identified in LNCaP a total of 11 genes differentially expressed ( FC > or= +/-1.5 ) . Eight of these genes , were overexpressed and three were underexpressed . Overexpressed genes included genes encoding for proteins involved in biosynthesis and metabolism of androgen ( P14061 ; P37058 ; P19099 ) , androgen receptor and androgen receptor co-regulators ( AR; P24385 ) , and signal transduction ( P04626 ; V- P62158 ; Q07889 ) whereas , underexpressed genes ( KLK3 ; P20151 ; Q15392 ) were androgen-regulated genes ( ARGs ) . No differentially expressed genes were scored in DU145 . Microarray data were confirmed by quantitative real-time PCR assay ( QRT-PCR ) . These data offer a selective genomic signature for dutasteride treatment in prostate epithelial cells and provide important insights in prostate cancer pathophysiology . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . Restriction of adenoviral replication to the transcriptional intersection of two different promoters for colorectal and pancreatic cancer treatment . In our current study , we developed oncolytic adenoviruses which preferentially lyse pancreatic and colon cancer cells by replacing viral E1 and/or E4 promoter with the tumor/tissue-specific promoters , cyclooxygenase-2 ( P35354 ) , midkine ( MK ) , or the cell cycle-dependent promoter , Q01094 . We generated three sets of recombinant adenoviral vectors . In the first set , only the native E1A promoter was replaced by the P35354 , MK , or Q01094 promoter , respectively . In the second set , the viral E4 promoter was substituted by these heterologous promoters and the viral E1A promoter was substituted by the ubiquitously active cytomegalovirus-IE promoter . In the third set , we substituted the viral E1A and E4 promoters with the P35354 , MK , or Q01094 promoter , respectively . In our system , transcriptional targeting of solitary viral E1A resulted in 50 % enhanced restricted vector replication when compared with an unrestricted replication-competent adenovirus . Furthermore , a targeted expression of the viral E1A gene products had a greater effect on restricted adenoviral replication than that of the E4 region . With our vectors , Ad. P36551 .MK and Ad.MK. P36551 , using two different heterologous promoters to control E1A and E4 expression , we showed enhanced viral replication specificity when compared with Ad. P36551 . P36551 or Ad.MK.MK , respectively . In a s.c. xenograft tumor model , there was no significant difference in the antineoplastic efficacy of the double heterologous promoter-controlled vectors when compared with our unrestricted replication-competent control adenovirus or vectors with only E1A transcriptionally driven by a heterologous promoter . Use of a cyclo-oxygenase 2 inhibitor for prophylaxis of cystoid macular oedema following cataract surgery : a randomized placebo-controlled trial . BACKGROUND : To assess the efficacy of Celecoxib , a cyclo-oxygenase 2 ( P35354 ) inhibitor , as prophylaxis for cystoid macular oedema after routine cataract surgery . METHODS : A prospective , randomized , double-blind placebo-controlled trial of 69 hospital patients undergoing cataract surgery . Celecoxib 200 mg twice daily or placebo was given immediately after surgery for 14 days . Optical coherence tomography was used to quantify macular thickness before surgery and on day 1 , week 2 and week 6 after surgery . RESULTS : Sixty-nine patients were enrolled , of which 33 received placebo and 36 received active drug . Clinically apparent cystoid macular oedema occurred in four of the treatment group and two of the placebo group ( P = 0.68 ) . No difference in best-corrected visual acuity was seen at 6 weeks ( P = 0.37 ) . Covariate analysis of the results at 2 weeks and 6 weeks showed a macular thickness of 3 % less in the treatment group compared with placebo ( P = 0.050 ) . CONCLUSION : Celecoxib may decrease macular thickening following routine cataract surgery at 2 and 6 weeks after surgery as measured by Stratus O75051 III . No difference in best-corrected visual acuity or clinically apparent cystoid macular oedema was seen . Further investigation of P35354 inhibitors in a larger prospective randomized trial is required . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . P11831 depletion affects the proliferation of the hepatocellular carcinoma cells HepG2 and JHH6 . For hepatocellular carcinoma ( HCC ) , a leading cause of cancer death world-wide , there is no effective therapy especially for the advanced stage of the disease . Thus , we started the investigations about a novel anti HCC approach based on the depletion of the transcription factor serum response factor ( P11831 ) in HCC cell lines ; P11831 choice was based on its recently proposed contribution to HCC tissue development and on its important role in cell proliferation . P11831 depletion , obtained by a siRNA ( siSRF797 ) , was studied in two HCC cell lines , i.e. HepG2 and JHH6 assigned to high and low hepatocytic differentiation grade on the base of the capacity to synthesize albumin . In the HCC cell lines examined , siSRF797 reduced both the mRNA and protein levels of P11831 without inducing unspecific interferon response or cytotoxicity . Moreover , P11831 depletion induced the reduction of S-phase cells and a decrease in cell number and vitality . Particularly in HepG2 , cell growth impairment was paralleled by the decrease of the levels of the transcription factor Q01094 together with some of its regulated genes . In HepG2 but not in JHH6 , P11831 depletion was associated with apoptosis . Finally , in both HepG2 and JHH6 , the combined administration of siSRF797 and bortezomib , a proteasome inhibitor whose therapeutic potential for HCC is considered attractive , further reduced cell viability compared to either siSRF797 or bortezomib treatment alone . In conclusion , P11831 depletion affects the expansion of the high and low differentiation grade HCC cells HepG2 and JHH6 . These results can pave the way to understand the role of P11831 in HCC development and possibly to identify novel anti HCC therapeutic strategies . 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 . Q96RP3 is expressed in human pregnant myometrial cells and regulates myosin light chain phosphorylation : potential role of the type-2 corticotropin-releasing hormone receptor in the control of myometrial contractility . The family of P06850 -related peptides are suggested to play important roles in the control of myometrial contractility during pregnancy and labor . In this study we investigated the expression of urocortin II ( P55089 II ) in human myometrium and its ability to phosphorylate intracellular components that can be involved in modulating myometrial contractility . Using RT-PCR and fluorescent in situ hybridization , we demonstrated that P55089 II and type-2 P06850 receptor ( Q13324 ) mRNAs were expressed in human nonpregnant and pregnant myometrium . Immunofluorescent studies confirmed protein expression of P55089 II in human pregnant myometrial cells , whereas chemical cross-linking studies with radiolabeled P55089 II confirmed the presence of Q13324 sites with an apparent molecular mass of 50 kDa . Treatment of primary human myometrial cells with P55089 II to specifically activate Q13324 resulted in a dose-dependent increase of myosin light chain ( MLC(20) ) phosphorylation . Activation of protein kinase C ( PKC ) and P27361 /2 was required for the P55089 II-induced activation of MLC(20) , because treatment of myometrial cells with inhibitors of MAPK kinase 1 ( U0126 ) and PKC ( bisindolylmaleimide ) inhibited the P55089 II-induced phosphorylation of MLC(20) . Furthermore , the P55089 II effect on MLC(20) was dependent on RhoA translocation to the membrane and subsequent activation of RhoA-associated kinase , as shown by the use of the specific inhibitors exoenzyme P01024 and Y27632 . Collectively , our data suggest a distinctive role for Q13324 - specific agonists like P55089 II in the control of myometrial contractility during human pregnancy involving sequential activation of PKC , MAPK kinase 1 , P27361 /2 , RhoA , and RhoA-associated kinase , leading to the MLC(20) phosphorylation . Distinct effects of inflammation on gliosis , osmohomeostasis , and vascular integrity during amyloid beta-induced retinal degeneration . In normal retinas , amyloid-β ( Aβ ) accumulates in the subretinal space , at the interface of the retinal pigment epithelium , and the photoreceptor outer segments . However , the molecular and cellular effects of subretinal Aβ remain inadequately elucidated . We previously showed that subretinal injection of Aβ(1-42) induces retinal inflammation , followed by photoreceptor cell death . The retinal Müller glial ( RMG ) cells , which are the principal retinal glial cells , are metabolically coupled to photoreceptors . Their role in the maintenance of retinal water/potassium and glutamate homeostasis makes them important players in photoreceptor survival . This study investigated the effects of subretinal Aβ(1-42) on RMG cells and of Aβ(1-42)-induced inflammation on retinal homeostasis . RMG cell gliosis ( upregulation of P14136 , vimentin , and nestin ) on day 1 postinjection and a proinflammatory phenotype were the first signs of retinal alteration induced by Aβ(1-42) . On day 3 , we detected modifications in the protein expression patterns of cyclooxygenase 2 ( P35354 ) , glutamine synthetase ( GS ) , Kir4.1 [ the inwardly rectifying potassium ( Kir ) channel ] , and aquaporin ( AQP ) -4 water channels in RMG cells and of the photoreceptor-associated P29972 . The integrity of the blood-retina barrier was compromised and retinal edema developed . Aβ(1-42) induced endoplasmic reticulum stress associated with sustained upregulation of the proapoptotic factors of the unfolded protein response and persistent photoreceptor apoptosis . Indomethacin treatment decreased inflammation and reversed the Aβ(1-42)-induced gliosis and modifications in the expression patterns of P35354 , Kir4.1 , and P29972 , but not of P55087 or GS . Nor did it improve edema . Our study pinpoints the adaptive response to Aβ of specific RMG cell functions . Expression of hypothalamic peptides in mice lacking neuronal nitric oxide synthase : reduced beta- P17813 immunoreactivity in the arcuate nucleus . The gas nitric oxide ( NO ) is an important messenger in brain signaling . Along with many other functions , NO is thought to influence the expression and/or release of various hypothalamic hormones ( corticotropin-releasing hormone ( P06850 ) , gonadotropin-releasing hormone ( DB00644 ) and vasopressin ) . To learn more about the role of NO in neuroendocrine mechanisms , we studied in mutant mice lacking neuronal isoform of NO synthase ( P29475 ) the cellular expression of P06850 , neurophysin ( the carrier protein of vasopressin/oxytocin ) and pro-opiomelanocortin ( P01189 ) , as well as of the P01189 -derived peptides beta-endorphin ( beta- P17813 ) , alpha-melanocyte-stimulating hormone ( alpha-MSH ) and corticotropin ( DB01285 ) by use of immunohistochemistry and in situ hybridization . Additionally , the remaining NO-generating capacities of the P29475 minus mice were investigated by NADPH-diaphorase histochemistry and citrulline immunohistochemistry as well as by immunohistochemical localization and Western blot analysis of endothelial NOS ( P29474 ) and P29475 isoforms . Amongst all hypothalamic peptides under investigation , only beta- P17813 was found to be altered in mutant mice . A morphometric analysis of beta- P17813 producing neurons of the arcuate nucleus revealed that significantly less cells were immunoreactive in mutant mice , whereas the expression of the precursor P01189 as well as of other P01189 -derived peptides was found to be unchanged . In addition to that , fewer beta- P17813 -immunoreactive fibers were found in the paraventricular nucleus of P29475 minus mice in comparison to wild-type animals . Hence , the reduction of hypothalamic beta- P17813 is probably a posttranslational event that might reflect a disturbed endorphinergic innervation of those hypothalamic neurons which normally express P29475 . In vivo magnetomotive optical molecular imaging using targeted magnetic nanoprobes . Dynamic magnetomotion of magnetic nanoparticles ( MNPs ) detected with magnetomotive optical coherence tomography ( MM- O75051 ) represents a new methodology for contrast enhancement and therapeutic interventions in molecular imaging . In this study , we demonstrate in vivo imaging of dynamic functionalized iron oxide MNPs using MM- O75051 in a preclinical mammary tumor model . Using targeted MNPs , in vivo MM- O75051 images exhibit strong magnetomotive signals in mammary tumor , and no significant signals were measured from tumors of rats injected with nontargeted MNPs or saline . The results of in vivo MM- O75051 are validated by Q9BWK5 , ex vivo MM- O75051 , DB06783 staining of histological sections , and immunohistochemical analysis of excised tumors and internal organs . The MNPs are antibody functionalized to target the human epidermal growth factor receptor 2 ( P04626 neu ) protein . Fc-directed conjugation of the antibody to the MNPs aids in reducing uptake by macrophages in the reticulo-endothelial system , thereby increasing the circulation time in the blood . These engineered magnetic nanoprobes have multifunctional capabilities enabling them to be used as dynamic contrast agents in MM- O75051 and Q9BWK5 . DB04630 stimulates vascular smooth muscle cell proliferation via big mitogen-activated protein kinase 1 activation . The nongenomic effects of aldosterone have been implicated in the pathogenesis of various cardiovascular diseases . DB04630 -induced nongenomic effects are attributable in part to the activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) , a classical mitogen-activated protein ( Q96HU1 ) kinase . Q13164 ( Q13164 ) , a newly identified Q96HU1 kinase , has been shown to be involved in cell proliferation , differentiation , and survival . We examined whether aldosterone stimulates Q13164 -mediated proliferation of cultured rat aortic smooth muscle cells ( RASMCs ) . P08235 ( MR ) expression and localization were evaluated by Western blotting analysis and fluorolabeling methods . P27361 /2 and Q13164 activities were measured by Western blotting analysis with the respective phosphospecific antibodies . Cell proliferation was determined by Alamar Blue colorimetric assay . DB04630 ( 0.1 to 100 nmol/L ) dose-dependently activated Q13164 in RASMCs , with a peak at 30 minutes . To clarify whether aldosterone-induced Q13164 activation is an MR-mediated phenomenon , we examined the effect of eplerenone , a selective MR antagonist , on aldosterone-induced Q13164 activation . DB00700 ( 0.1 to 10 micromol/L ) dose-dependently inhibited aldosterone-induced Q13164 activation in RASMCs . DB04630 also stimulated RASMC proliferation , which was inhibited by eplerenone . DB04630 -mediated phenomena were concluded to be attributable to a nongenomic effect because cycloheximide failed to inhibit aldosterone-induced Q13164 activation . Transfection of dominant-negative Q96HU1 kinase/ P29323 kinase 5 ( Q13163 ) , which is an upstream regulator of Q13164 , partially inhibited aldosterone-induced RASMC proliferation , which was almost completely inhibited by MEK inhibitor PD98059 . In addition to the classical steroid activity , rapid nongenomic effects induced by aldosterone may represent an alternative etiology for vascular diseases such as hypertension . Role of cardiovascular aldosterone in hypertension . DB04630 plays an important role in the pathogenesis of cardiovascular disease . We have reported that aldosterone is synthesized in cardiovascular tissues and local aldosterone synthesis plays important roles for hypertension and cardiac hypertrophy . High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP . Plasma aldosterone concentrations and P06703 were decreased by high salt intake in SHRSP . DB04630 production , the expression of P19099 mRNA and angiotensin II receptor AT1R mRNA in blood vessels were significantly increased by high salt intake . These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the vascular tissue in SHRSP , which may contribute to the development of malignant hypertension in salt-loaded SHRSP . However , there are several reports of conflicting data . P08235 ( MR ) binding is tightly regulated by the enzyme 11beta-hydroxysteroid dehydrogenase type 2 ( 11beta-HSD2 ) which selectively metabolizes glucocorticoids to inactive metabolites , thus allowing for MR activation by aldosterone . We have reported that decreased 11beta-HSD2 in blood vessels in Dahl salt-sensitive ( DS ) rats , a model for salt-sensitive hypertension . Local aldosterone excess may play a significant role in the salt sensitivity and development of hypertension . High sodium intake decreased circulating rennin-angiotensin-aldosterone system and increased blood pressure and cardiac hypertrophy in DS rats , which were prevented by the treatment with a selective MR antagonist , eplerenone . DB00700 also improved endothelial nitric oxide synthase ( P29474 ) activity and P29474 mRNA expression in blood vessels in DS rats . These results further suggest that not only circulating aldosterone but also local aldosterone is of critical importance in the pathophysiology of cardiovascular disorders . Cell fate determination factor Q9UI36 inhibits c-Jun-induced contact-independent growth . The cell fate determination factor Q9UI36 plays a key role in cellular differentiation in metazoans . Q9UI36 is engaged in multiple context-dependent complexes that activate or repress transcription . Q9UI36 can be recruited to DNA via the Six1/Eya bipartite transcription ( DNA binding/coactivator ) complex . c-Jun is a critical component of the activator protein ( AP ) -1 transcription factor complex and can promote contact-independent growth . Herein , Q9UI36 inhibited c-Jun-induced DNA synthesis and cellular proliferation . Excision of c-Jun with Cre recombinase , in c-jun(f1/f1) 3T3 cells , abrogated Q9UI36 -mediated inhibition of DNA synthesis . c-Jun expression rescued Q9UI36 -mediated inhibition of cellular proliferation . Q9UI36 inhibited induction of c-Jun by physiological stimuli and repressed c-jun target genes ( cyclin A , beta-PAK , and stathmin ) . Q9UI36 bound c-Jun and inhibited AP-1 transcriptional activity . c-jun and c-fos were transcriptionally repressed by Q9UI36 , requiring the conserved N-terminal ( dac and ski/sno [ DS ] ) domain . c-fos transcriptional repression by Q9UI36 requires the P11831 site of the c-fos promoter . Q9UI36 inhibited c-Jun transactivation through the delta domain of c-Jun . Q9UI36 coprecipitated the histone deacetylase proteins ( Q13547 , Q92769 , and NCoR ) , providing a mechanism by which Q9UI36 represses c-Jun activity through the conserved delta domain . An oncogenic v-Jun deleted of the delta domain was resistant to Q9UI36 repression . Collectively , these studies demonstrate a novel mechanism by which Q9UI36 blocks c-Jun-mediated contact-independent growth through repressing the c-Jun delta domain . P08235 antagonism in the treatment of chronic central serous chorioretinopathy : a pilot study . PURPOSE : Based on experimental data showing that central serous chorioretinopathy could result from overactivation of mineralocorticoid receptor pathway in choroid vessels , the authors studied eplerenone , a mineralocorticoid receptor antagonist , as a potential treatment for chronic central serous chorioretinopathy . METHODS : This nonrandomized pilot study included 13 patients with central serous chorioretinopathy of at least 4-month duration , treated with 25 mg/day of oral eplerenone for a week followed by 50 mg/day for 1 or 3 months . The primary outcome measure was the changes in central macular thickness recorded by optical coherence tomography , and the secondary outcomes included changes in foveal subretinal fluid ( P11831 ) measured by O75051 , in best-corrected visual acuity ( BCVA ) and the percentage of eyes achieving complete resolution of subretinal fluid during the treatment period . RESULTS : Central macular thickness decreased significantly from 352 ± 139 μm at baseline to 246 ± 113 μm and 189 ± 99 μm at 1 and 3 months under eplerenone treatment ( P < 0.05 and P < 0.01 , respectively ) . At 3 months , the subretinal fluid significantly decreased compared with baseline subretinal fluid ( P < 0.01 ) and best-corrected visual acuity significantly improved compared with baseline best-corrected visual acuity ( P < 0.001 ) . CONCLUSION : DB00700 treatment was associated with a significant reduction in central macular thickness , subretinal fluid level , and an improvement in visual acuity . Randomized controlled trials are needed to confirm these encouraging results . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . DB00104 is the favorable alternative for cisplatin resistance reversal of ovarian cancer in vitro and in nude mice in vivo . This study aimed to observe the effects of octreotide ( O75051 ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo . MTT method and flow cytometry were used to investigate the effect of cisplatin , O75051 or the combination of these two compounds on the proliferation and apoptosis of SKOV3- O60220 cells . The size and weight of xenograft tumors from the nude mice model were measured . Real-time PCR was used to detect the mRNA expression of P30874 , P08183 , Q92887 , Q86UG4 -pi and P00533 in SKOV3/ O60220 cells following the different treatment . At the concentration of 2.5-20 g/ml , O75051 significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3- O60220 cells ' response to cisplatin . Unchanged expression was found in P30874 on the SKOV3/ O60220 cell in vitro after O75051 treatment , but increased expression in vivo ( p < 0.05 ) . O75051 increased Q86UG4 -pi expression ( p < 0.05 ) and reduced Q92887 and P00533 expression ( p < 0.05 ) in a dose-dependent manner . The similar results were obtained in mice in vivo experiment , except the reduced expression of Q86UG4 -pi . It is suggested that O75051 could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface P30874 , and reverse cisplatin resistance through inhibition of Q92887 , P00533 , and even Q86UG4 -pi expressions . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . P04406 mediates nitrosylation of nuclear proteins . S-nitrosylation of proteins by nitric oxide is a major mode of signalling in cells . S-nitrosylation can mediate the regulation of a range of proteins , including prominent nuclear proteins , such as Q92769 ( ref. 2 ) and P09874 ( ref. 3 ) . The high reactivity of the nitric oxide group with protein thiols , but the selective nature of nitrosylation within the cell , implies the existence of targeting mechanisms . Specificity of nitric oxide signalling is often achieved by the binding of nitric oxide synthase ( NOS ) to target proteins , either directly or through scaffolding proteins such as P78352 ( ref. 5 ) and O75052 . As the three principal isoforms of NOS -- neuronal NOS ( P29475 ) , endothelial NOS ( P29474 ) and inducible NOS ( P35228 ) -- are primarily non-nuclear , the mechanisms by which nuclear proteins are selectively nitrosylated have been elusive . P04406 ( P04406 ) is physiologically nitrosylated at its DB00151 150 residue . Nitrosylated P04406 ( SNO- P04406 ) binds to Siah1 , which possesses a nuclear localization signal , and is transported to the nucleus . Here , we show that SNO- P04406 physiologically transnitrosylates nuclear proteins , including the deacetylating enzyme sirtuin-1 ( Q96EB6 ) , histone deacetylase-2 ( Q92769 ) and DNA-activated protein kinase ( DNA-PK ) . Our findings reveal a novel mechanism for targeted nitrosylation of nuclear proteins and suggest that protein-protein transfer of nitric oxide groups may be a general mechanism in cellular signal transduction . Dynamic simulations of pathways downstream of P00533 -family , including mutations and treatments : concordance with experimental results . The pathways downstream of ErbB-family proteins are very important in BC , especially when considering treatment with onco-protein inhibitors . We studied and implemented dynamic simulations of four downstream pathways and described the fragment of the signaling network we evaluated as a Molecular Interaction Map . Our simulations , enacted using Ordinary Differential Equations , involved 242 modified species and complexes , 279 reversible reactions and 111 catalytic reactions . Mutations within a single pathway tended to be mutually exclusive ; only inhibitors acting at , or downstream ( not upstream ) , of a given mutation were active . A double alteration along two distinct pathways required the inhibition of both pathways . We started an analysis of sensitivity/robustness of our network , and we systematically introduced several individual fluctuations of total concentrations of independent molecular species . Only very few cases showed significant sensitivity . We transduced the ErbB2 over-expressing BC line , BT474 , with the P01112 ( V12 ) mutant , then treated it with ErbB-family and phosphorylated MEK ( MEKPP ) inhibitors , DB01259 and U0126 , respectively . Experimental and simulation results were highly concordant , showing statistical significance for both pathways and for two respective endpoints , i.e. phosphorylated active forms of P29323 and Akt , p one tailed = .0072 and = .0022 , respectively . Working with a complex 39 basic species signaling network region , this technology facilitates both comprehension and effective , efficient and accurate modeling and data interpretation . Dynamic network simulations we performed proved to be both practical and valuable for a posteriori comprehension of biological networks and signaling , thereby greatly facilitating handling , and thus complete exploitation , of biological data . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Epigenetic mechanisms in the dopamine D2 receptor-dependent inhibition of the prolactin gene . Transcription of the prolactin gene is dynamically controlled by positive and negative hormone signals that target the regulatory promoter region . Based on the inducibility of prolactin gene expression by inhibitors of histone deacetylases ( HDACs ) , we examined the role of histone acetylation at the genomic prolactin promoter as a late step in transcriptional regulation . Chromatin immunoprecipitation analysis of GH4 cells revealed elevated levels of acetylated histones in the promoter and enhancer regions of the gene , compared with downstream intron sequences . 17beta-Estradiol stimulated histone H4 acetylation in the promoter region by 2- to 3-fold within 30 min . Dopamine inhibited histone H4 acetylation by 2-fold in 30 min , an effect mimicked by the MAPK kinase ( Q02750 ) inhibitor U0126 . In contrast , the synthetic glucocorticoid dexamethasone , which inhibits prolactin transcription , failed to alter histone acetylation over the same time frame . Association of transcription activator Pit-1 with the prolactin promoter was unchanged by hormone treatment . However , in response to dopamine , histone deacetylase Q92769 and corepressor mSin3A were rapidly recruited to the prolactin promoter , and association was sustained above basal levels over a 1-h period . Consistent with this corepressor function , depletion of endogenous mSin3A by small interfering RNA was sufficient to enhance prolactin gene expression by 70 % , comparable to the induction by the HDAC inhibitor , trichostatin A . These studies demonstrate that dopamine D2 receptor activation and inhibition of MAPK ( P27361 /2 ) signaling lead to rapid deacetylation of histones at the genomic prolactin promoter . Recruitment of specific HDAC/ corepressor complexes may be an important mechanism for repression of target gene transcription by Gi/o-coupled receptors . P29972 in the peritoneal membrane : implications for peritoneal dialysis and endothelial cell function . PD ( peritoneal dialysis ) is an established mode of renal replacement therapy , based on the exchange of fluid and solutes between blood in peritoneal capillaries and a dialysate that has been introduced into the peritoneal cavity . The dialysis process involves diffusive and convective transports and osmosis through the PM ( peritoneal membrane ) . Computer simulations predicted that the PM contains ultrasmall pores ( radius < 3 A , 1 A=10(-10) m ) , responsible for up to 50 % of UF ( ultrafiltration ) , i.e. the osmotically driven water movement during PD . Several lines of evidence suggest that P29972 ( aquaporin-1 ) is the ultrasmall pore responsible for transcellular water permeability during PD . Treatment with corticosteroids induces the expression of P29972 in the PM and improves water permeability and UF in rats without affecting the osmotic gradient and permeability for small solutes . Studies in knockout mice provided further evidence that osmotically driven water transport across the PM is mediated by P29972 . P29972 and P29474 ( endothelial nitric oxide synthase ) show a distinct regulation within the endothelium lining the peritoneal capillaries . In acute peritonitis , the up-regulation of P29474 and increased release of nitric oxide dissipate the osmotic gradient and prevent UF , whereas P29972 expression is unchanged . These results illustrate the usefulness of the PM to investigate the role and regulation of P29972 in the endothelium . The results also emphasize the critical role of P29972 during PD and suggest that manipulation of P29972 expression may be used to increase water permeability across the PM . DB01259 -mediated cyclooxygenase-2 expression via epidermal growth factor receptor/ Q15717 interaction enhances the aggressiveness of triple-negative breast cancer cells . DB01259 , a dual epidermal growth factor receptor ( P00533 ) /human epidermal growth factor receptor 2 ( P04626 ) kinase inhibitor , showed clinical benefits in advanced P04626 -positive breast cancer patients . Because some triple-negative breast cancers ( TNBCs ) frequently overexpress P00533 , the antitumor activity of lapatinib in such diseases was also tested . However , the results showed a worse event-free survival rate . It remains unknown whether and how lapatinib elicits the aggressiveness of such cancer cells . In this study , our results demonstrated that lapatinib facilitated axillary and lung metastases of triple-negative MDA-MB-231 breast cancer cells without affecting their viability , leading to worse survival in orthotopic xenograft mice . The lapatinib-increased motility was attributed by the elevation of P00533 through the downregulation of microRNA-7 and by the subsequent overexpression of cyclooxygenase-2 ( P35354 ) . Strikingly , independent of its kinase activity , the elevated P00533 at least partly stabilized P35354 expression by enhancing the binding of Q15717 to P35354 mRNA . Our results suggest that lapatinib may increase the migration and invasion of MDA-MB-231 cells by upregulating P00533 and P35354 through the downregulation of microRNA-7 , providing a potential explanation for the worse clinical outcome of TNBC patients who receive lapatinib-based treatment . These findings also shed new light on the molecular mechanism of P35354 mRNA stabilization by P00533 in a kinase-independent manner . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . 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 . Effect of cyclooxygenase inhibitors on the P06850 -induced pituitary-adrenocortical activity during crowding stress . The aim of the present study was to determine the effect of social stress and significance of prostaglandins ( PG ) generated by constitutive and inducible cyclooxygenase ( P23219 and P35354 ) in the stimulation of hypothalamic-pituitary-adrenal ( Q9Y251 ) axis by corticotropin releasing hormone ( P06850 ) under basal and social crowding stress conditions . The stressed rats were crowded in groups of 24 to a cage for 3 or 7 days , whereas the control animals were haused in groups of 7 to a cage of the same size . The activity of Q9Y251 axis was determined by measuring plasma DB01285 and serum corticosterone levels 1 h after i.p . P06850 administration . Inhibitors of P23219 , piroxicam ( 0.2 , 2.0 , and 5.0 mg/kg ) , and P35354 , compound NS-398 ( 0.2 and 2.0 mg/kg ) , were administered i.p . 15 min prior to P06850 ( 0.1 microg/kg i.p. ) to control or crowded rats . The obtained results indicate that social stress for 3 and 7 days markedly intensifies the stimulatory action of P06850 on DB01285 secretion . Neither piroxicam nor NS-398 induce any significant effect on the P06850 -elicited DB01285 and corticosterone secretion in non-stressed or crowded rats . Therefore , PG generated by P23219 or P35354 do not participate to a significant extent in the stimulation of Q9Y251 axis by P06850 under either basal conditions or during crowding stress . These results also indicate that the stimulatory action of P06850 on DB01285 secretion is not only completely resistant to desensitization but is sensitized during social crowding stress . The results contrast with a significant involvement of PG in the vasopressin-induced stimulation of Q9Y251 response during crowding stress . Interleukin-1β causes excitotoxic neurodegeneration and multiple sclerosis disease progression by activating the apoptotic protein p53 . BACKGROUND : Understanding how inflammation causes neuronal damage is of paramount importance in multiple sclerosis ( MS ) and in other neurodegenerative diseases . Here we addressed the role of the apoptotic cascade in the synaptic abnormalities and neuronal loss caused by the proinflammatory cytokines interleukin-1β ( IL-1β ) and tumor necrosis factor ( P01375 -α ) in brain tissues , and disease progression caused by inflammation in relapsing-remitting MS ( RRMS ) patients . RESULTS : The effect of IL-1β , but not of P01375 -α , on glutamate-mediated excitatory postsynaptic currents was blocked by pifithrin-α ( DB00522 ) , inhibitor of p53 . The protein kinase C ( PKC ) /transient receptor potential vanilloid 1 ( Q8NER1 ) pathway was involved in IL-1β-p53 interaction at glutamatergic synapses , as pharmacological modulation of this inflammation-relevant molecular pathway affected DB00522 effects on the synaptic action of IL-1β . IL-1β-induced neuronal swelling was also blocked by DB00522 , and IL-1β increased the expression of P38936 , a canonical downstream target of activated p53.Consistent with these in vitro results , the Pro/Pro genotype of p53 , associated with low efficiency of transcription of p53-regulated genes , abrogated the association between IL-1β cerebrospinal fluid ( P04141 ) levels and disability progression in RRMS patients . The interaction between p53 and P04141 IL-1β was also evaluated at the optical coherence tomography ( O75051 ) , showing that IL-1β-driven neurodegenerative damage , causing alterations of macular volume and of retinal nerve fibre layer thickness , was modulated by the p53 genotype . CONCLUSIONS : Inflammatory synaptopathy and neurodegeneration caused by IL-1β in RRMS patients involve the apoptotic cascade . Targeting IL-1β-p53 interaction might result in significant neuroprotection in MS . P01375 inhibits flow and insulin signaling leading to NO production in aortic endothelial cells . Endothelial cells release nitric oxide ( NO ) acutely in response to increased " flow " or fluid shear stress ( FSS ) , and the increase in NO production is correlated with enhanced phosphorylation and activation of endothelial nitric oxide synthase ( P29474 ) . Both vascular endothelial growth factor and FSS activate endothelial protein kinase B ( P31749 ) by way of incompletely understood pathway(s) , and , in turn , P31749 phosphorylates P29474 at DB00133 -1179 , causing its activation . In this study , we found that either FSS or insulin stimulated insulin receptor substrate-1 ( P35568 ) tyrosine and serine phosphorylation and increased P35568 -associated phosphatidylinositol 3-kinase activity , phosphorylation of P31749 DB00133 -473 , phosphorylation of P29474 DB00133 -1179 , and NO production . Brief pretreatment of bovine aortic endothelial cells with tumor necrosis factor-alpha ( P01375 ) inhibited the above described FSS- or insulin-stimulated protein phosphorylation events and almost totally inhibited FSS- or insulin-stimulated NO production . These data indicate that FSS and insulin regulate P29474 phosphorylation and NO production by overlapping mechanisms . This study suggests one potential mechanism for the development of endothelial dysfunction in disease states with alterations in insulin regulation and increased P01375 levels . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . DB00227 induces the expression of bradykinin type 2 receptors in cultured human coronary artery endothelial cells . Cardioprotective bradykinin type-2 receptors ( BK-2Rs ) are downregulated in the myocardial endothelium of both human and rat failing hearts . Statins are cardioprotective drugs that reduce the level of plasma cholesterol but also exert cholesterol-independent pleiotropic effects . Here we examined the effect of lovastatin on BK-2R expression in cultured human coronary artery endothelial cells . The effect of lovastatin on the expression of BK receptors in human coronary artery endothelial cells ( HCAECs ) was examined by real-time PCR , Western blot analysis and immunocytochemistry . DB00227 induced a time- and concentration-dependent increase in both BK-2R and BK-1R mRNA expression in the cultured HCAECs . Also , the number of functional BK-2Rs capable of inducing BK-mediated NO production and cGMP signaling was increased in the lovastatin-treated HCAECs . Mevalonate , the direct metabolite of P04035 , reversed the effect of lovastatin . Furthermore , lovastatin inhibited Rho activation and a selective inhibitor of Rho-associated kinases , Y-27632 , induced a similar increase in BK-2R expression as lovastatin . In contrast , a specific inhibitor of P35354 , NS398 , significantly inhibited the lovastatin-induced expression of BK-2Rs . Here we show for the first time that lovastatin induces the expression of BK-2Rs in cultured human coronary artery endothelial cells through a novel cholesterol-independent pleiotropic mechanism that involves RhoA kinase inhibition and P35354 activation . Thus , reported beneficial effects of statins in cardiovascular diseases may be partly mediated by an increased expression of cardioprotective BK-2Rs in the endothelial cells of the coronary tree . Moreover , the use of P35354 inhibitors may affect the level of endothelial BK-2Rs in a negative fashion . Expression of aquaporin1 , 3 , and 4 , P55011 , and Q13621 in the human endolymphatic sac . OBJECTIVE : To locate aquaporin ( AQP ) 1 , 3 , and 4 , Na-K-2Cl cotransporter ( NKCC ) 1 and 2 in the human endolymphatic sac ( ES ) . METHODS : A sample of human ES was harvested during the removal of vestibular schwannoma via the translabyrinthine approach . The sample was immediately fixed in 4 % paraformaldehyde and embedded in O75051 compound . Immunohistochemistry was performed with P29972 , 3 , and 4 , P55011 , and Q13621 polyclonal antibodies . RESULTS : P29972 , Q92482 , and Q13621 were strongly expressed in the epithelial layer of the ES . P55087 and P55011 were weakly expressed in the epithelial layer of the ES . CONCLUSIONS : As it is impossible to perform quantitative analysis based on the fluorescence intensity of each immunoreactivity , we have presented the existence of P29972 , 3 , and 4 , P55011 , and Q13621 in the ES . The expression of P55011 and 2 indicated that the ES may have both secretory and adsorptive functions to maintain the homeostasis of endolymph . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Hyperosmolar mannitol simulates expression of aquaporins 4 and 9 through a p38 mitogen-activated protein kinase-dependent pathway in rat astrocytes . The membrane pore proteins , aquaporins ( AQPs ) , facilitate the osmotically driven passage of water and , in some instances , small solutes . Under hyperosmotic conditions , the expression of some AQPs changes , and some studies have shown that the expression of P29972 and P55064 is regulated by MAPKs . However , the mechanisms regulating the expression of P55087 and AQP9 induced by hyperosmotic stress are poorly understood . In this study , we observed that hyperosmotic stress induced by mannitol increased the expression of P55087 and AQP9 in cultured rat astrocytes , and intraperitoneal infusion of mannitol increased P55087 and AQP9 in the rat brain cortex . In addition , a p38 MAPK inhibitor , but not P29323 and JNK inhibitors , suppressed their expression in cultured astrocytes . AQPs play important roles in maintaining brain homeostasis . The expression of P55087 and AQP9 in astrocytes changes after brain ischemia or traumatic injury , and some studies have shown that p38 MAPK in astrocytes is activated under similar conditions . Since mannitol is commonly used to reduce brain edema , understanding the regulation of AQPs and p38 MAPK in astrocytes under hyperosmotic conditions induced with mannitol may lead to a control of water movements and a new treatment for brain edema .

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Absence of p21Waf1/Cip1/Sdi1 modulates macrophage differentiation and inflammatory response and protects against atherosclerosis . BACKGROUND : The tumor suppressor p53 protects against atherosclerosis progression in several different mouse models . A major target of p53 is P38936 , the cyclin-dependent kinase inhibitor that regulates entry into the cell cycle of different types of cells , including stem cells . P38936 is also involved in the maturation and differentiation of monocytes into macrophages . METHODS AND RESULTS : We studied the effect of p21Waf1 inactivation on atherosclerosis development in apolipoprotein E-deficient mice ( apoE-/- ) . Contrary to previous data suggesting a protective role for P38936 , we found that absence of P38936 , either globally or in bone marrow-derived cells , protects against atherosclerosis . Atherosclerotic lesions of P38936 -/-/apoE-/- mice exhibit a more stable phenotype , with increased apoptosis and reduced inflammatory vascular cell adhesion molecule-1 immunostaining but no difference in cellular proliferation compared with lesions of P38936 +/+/apoE-/- mice . Because bone marrow-derived cells mediate many of the effects of P38936 , we examined the expression profile of 23 genes in macrophages using real-time polymerase chain reaction . Compared with their P38936 +/+ counterparts , peritoneal macrophages of P38936 -/- mice express lower levels of proinflammatory markers , including macrophage inflammatory proteins 1 and 2 and interleukin-1alpha , and higher levels of putative protective genes , such as scavenger receptor type B-I and P01130 -related protein . Furthermore , we found that , in comparison with P38936 +/+ macrophages , P38936 -/- macrophages displayed increased phagocytic activity toward fluorescent latex microspheres as well as apoptotic cells , thus uncovering a novel mechanism of the antiinflammatory activity of P38936 -/- macrophages . CONCLUSIONS : Loss of P38936 protects against atherosclerosis in apoE-/- mice . The data underscore the important role of P38936 in macrophage function and inflammation and provide insight into the mechanism of the proatherogenic effect of P38936 . 3-Hydroxy-3-methylglutaryl coenzyme a reductase and isoprenylation inhibitors induce apoptosis of vascular smooth muscle cells in culture . Recent evidence suggests that apoptosis may be involved in the control of vascular smooth muscle cell ( VSMC ) number in atherosclerotic lesions . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors have been reported to induce apoptosis in a variety of tumor cell lines . To evaluate whether these agents also induce apoptosis of VSMCs , cultured rat VSMCs were treated with increasing doses of atorvastatin in the presence of FBS as a survival factor . The presence of apoptosis was evaluated by morphological criteria , annexin V binding , and DNA fragmentation and quantified as the proportion of hypodiploid cells by flow cytometry . DB01076 induced apoptosis in a dose-dependent manner , an effect also seen with simvastatin and lovastatin , but not with the hydrophilic drug pravastatin . The proapoptotic effect of statins was seen only when the inhibition of acetate incorporation into sterols was > 95 % and was fully reversed by mevalonate , farnesyl pyrophosphate , and geranylgeranyl pyrophosphate but not by isopentenyl adenosine , ubiquinone , or squalene , suggesting a role for prenylated proteins in the regulation of VSMC apoptosis . To further assess the role of protein prenylation , VSMCs were exposed to the prenyl transferase inhibitors perillic acid and manumycin A . Both agents induced VSMC apoptosis as evaluated by the above-mentioned criteria . Finally , VSMC treatment with lipophilic statins was associated with decreased prenylation of P38936 -Rho B , further supporting the role of protein prenylation inhibition in statin-induced VSMC apoptosis . The present data suggest that interference with protein prenylation by P04035 inhibitors or other agents may provide new strategies for the prevention of neointimal thickening . P01308 generates free radicals in human fibroblasts ex vivo by a protein kinase C-dependent mechanism , which is inhibited by pravastatin . P01308 can generate oxygen free radicals . Statins , P04035 inhibitors , exert a powerful antioxidant effect . The present study aimed to clarify the mechanisms through which insulin generates free radicals and to assess whether pravastatin modulates such effects . In cultured skin fibroblasts from human volunteers exposed to high insulin concentration , either in the presence or in the absence of pravastatin , insulin induced translocation of the p47(phox) subunit of NAD(P)H oxidase from the cytosol to the membrane and generation of radical oxygen species through a PKC delta-dependent mechanism . The insulin-induced translocation of p47(phox) was PKC delta dependent and attenuated by pravastatin , but independent of the activation of Akt and Rac1 . P01308 -induced Akt phosphorylation was increased by pravastatin and P27361 /2 phosphorylation attenuated . The present study demonstrates a novel mechanism by which insulin stimulates the generation of free radicals in human fibroblasts , ex vivo . It involves phosphatidylinositol 3-kinase , PKC delta , and p47(phox) translocation and promotes P27361 /2 phosphorylation . DB00175 inhibited radical oxygen species production by inhibiting PKC delta . These observations offer a robust explanation for the positive effects of pravastatin treatment in patients with insulin resistance syndrome . The natriuretic peptide neurohormonal system modulation by vasopeptidase inhibitors -- the novel therapeutical approach of hypertension treatment . Vasopeptidase inhibitors ( VPI ) are a new promising class of drugs , that simultaneously inhibit Angiotensin - Converting Enzyme ( P12821 ) and an enzyme Neutral Endopeptidase ( NEP ) , that cleaves the natriuretic peptides . These drugs , such as omapatrilat , sampatrilat , fasidotrilat , by combined inhibition of P12821 and degradation of natriuretic peptides and in turn by inhibiting the P00797 - Angiotensin - DB04630 system and potentiating the Natriuretic Peptide system and Kinin system should decrease the mortality rate in the group of patients with hypertension being not adequately controlled with P12821 inhibitors . Thus , finding the new therapeutic strategy using drugs that act on the hormonal systems other than P00797 - Angiotensin - DB04630 system seems to be crucial . The aim of the study was to compare the molecular aspects of the conventional schemes that are being used in the antihypertension therapy to the new drugs from the vasopeptidase inhibitors group -- with focusing on the natriuretic peptide system ( P0C0P6 ) -- and , taking these considerations , making clues about therapeutical implications to reveal promising results in antihypertension treatment . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . DB01373 -sensing receptor gene polymorphism Arg990Gly and its possible effect on response to cinacalcet HCl . DB01012 , a novel calcimimetic compound , is effective in reducing parathyroid hormone ( PTH ) levels in approximately 70 % of patients with secondary hyperparathyroidism . However , interindividual variations in the dose required to achieve the treatment goal have been noted in clinical studies . Our investigation examined the genetic polymorphisms of the calcium-sensing receptor ( P41180 ) gene as one possible cause of the different responses to cinacalcet . We report data on seven end-stage renal failure patients who were treated with regular haemodialysis and who participated in clinical trials of cinacalcet . All patients had secondary hyperparathyroidism with baseline intact PTH ( iPTH ) levels greater than 600 pg/ml . Three patients were male and four female with a mean+/-SD age of 60+/-12 years . DNA was extracted from peripheral lymphocytes . An area in exon 7 of the P41180 gene was amplified by the polymerase chain reaction and sequenced . Mean+/-SD baseline iPTH was 1086+/-189 pg/ml . The five patients without Arg990Gly demonstrated a 29.7+/-4.0 % ( +/-SEM ) reduction in iPTH from individual baseline . One patient was found to be homozygous for the Arg990Gly polymorphism and another was heterozygous for both arginine and glycine alleles . The homozygous patient showed a significantly higher sensitivity to cinacalcet compared to the other patients ( P=0.003 ) with a 76.3+/-7.7 % reduction in iPTH from baseline . No polymorphisms were noted in codons 986 or 1011 . This preliminary study points to the possibility that patients homozygous for glycine at the 990 position in exon 7 of the P41180 may be more sensitive to the calcimimetic drug cinacalcet compared to those who are homozygous for arginine at that location . New insights into the role of calcium-sensing receptor activation . The discovery of the calcium-sensing receptor ( P41180 ) prompted the identification of substances that affect its function . DB01012 , for example , is a drug that allosterically modifies the receptor so as to increase its sensitivity to circulating calcium ( thus the name " calcimimetic " ) and in this way decreases parathyroid hormone secretion . Clinical use of cinacalcet is already approved for the treatment of primary and secondary hyperparathyroidism , but research is ongoing to identify further potential actions of this drug . The effects and functions of the P41180 have been evaluated in different systems and tissues , beyond parathyroid glands , such arterial walls . A complete understanding of the properties of calcimimetics are of obvious clinical interest , since therapeutic indications may be affected accordingly . DB01012 Treatment in an Adolescent With Concurrent 22q11.2 Deletion Syndrome and Familial Hypocalciuric Hypercalcemia Type 3 Caused by P53680 Mutation . CONTEXT : The 22q11.2 deletion syndrome ( DS ) is a common multiple anomaly syndrome in which typical features include congenital heart defects , facial dysmorphism , and palatal anomalies . Hypocalcemia due to hypoparathyroidism is a common endocrine manifestation resulting from variable parathyroid hypoplasia , but hypercalcemia has not previously been reported in 22q11.2 DS . CASE DESCRIPTION : Our patient is a 16-year-old adolescent male with dysmorphic facial features and delayed motor and speech development . At 2 years of age , 22q11.2 DS was confirmed by fluorescence in situ hybridization . In contrast to hypoparathyroidism that is usually seen in 22q11.2 DS , this patient had early childhood-onset hypercalcemia with inappropriately high PTH levels and hypocalciuria . Genomic DNA was obtained from the proband and screened for calcium-sensing receptor ( P41180 ) mutations with negative results . No parathyroid tissue could be localized by imaging or surgical exploration . As a result of symptomatic hypercalcemia , the patient was treated with a calcimimetic ( cinacalcet ) . During the treatment , plasma calcium normalized with mild symptoms of hypocalcemia . After discontinuation of cinacalcet , calcium returned to high pretreatment levels . Further DNA analysis of adaptor protein-2 σ subunit ( P53680 ) showed a heterozygous missense mutation c.44 G > T , resulting in a p.R15L substitution ; the mutation was absent in the healthy parents and two siblings . CONCLUSIONS : Hypercalcemia in our patient with 22q11.2 DS could be explained by the de novo mutation in P53680 . Identification of a genetic cause for hypercalcemia is helpful in guiding management and avoiding unnecessary treatment . [ Vascular Calcification - Pathological Mechanism and Clinical Application - . The effect of cinacalcet on vascular calcification ] . DB01012 acts on calcium receptors ( CaR ) expressed on chief cells of the parathyroid gland to inhibit the secretion of parathyroid hormone ( PTH ) . This drug inhibits PTH secretion without causing an elevation of serum calcium and phosphorus , unlike active vitamin D . Several experimental studies demonstrated an inhibitory effect of calcimimetics on the progression of vascular calcification in animals with chronic kidney disease ( CKD ) , in keeping with the expression of the calcium-sensing receptor ( P41180 ) in vascular tissue . The EVOLVE , evaluated in patients with CKD 5D the effects of the cinacalcet on the progression of vascular calcification and hard cardiovascular outcomes , respectively . The EVOLVE trials missed their respective primary end point by intent-to-treat analysis . However , recently , in order to define the frequency of fatal and nonfatal cardiovascular events attributable to atherosclerotic and nonatherosclerotic mechanisms , risk factors for these events , and the effects of cinacalcet , post hoc analysis using adjudicated data collected during the EVOLVE Trial were perfomed . In this trial , combining fatal and nonfatal cardiovascular events , randomization to cinacalcet reduced the rates of sudden death and heart failure . Patients randomized to cinacalcet experienced fewer nonatherosclerotic cardiovascular events , while the effect of cinacalcet on atherosclerotic events did not reach statistical significance . Impact of clinically relevant mutations on the pharmacoregulation and signaling bias of the calcium-sensing receptor by positive and negative allosteric modulators . DB01012 is predominantly used to treat secondary hyperparathyroidism due to end-stage renal failure , but , more recently , its potential clinical efficacy in treating patients with loss-of-function mutations in the calcium-sensing receptor ( P41180 ) has been recognized . Many clinically relevant P41180 mutations are located in the heptahelical membrane spanning and extracellular loop regions of the receptor , where allosteric modulators are predicted to bind . The aim of the present study was to investigate the impact of such mutations on the pharmacoregulation of the P41180 by the positive and negative allosteric modulators , cinacalcet and P0C0P6 -2143 , respectively . Both cinacalcet and P0C0P6 -2143 effectively rescued mutants whose cell surface expression was substantially impaired , suggesting that both classes of drug can stabilize a receptor conformation that is trafficked more effectively to the cell surface . In addition , functional impairments in almost all mutant CaSRs were rescued by either cinacalcet or P0C0P6 -2143 via restoration of intracellular signaling . There was a significantly greater ability of both compounds to modulate agonist-stimulated intracellular Ca(2+) mobilization than P27361 /2 phosphorylation , indicating that the allosteric modulators engender bias in agonist-stimulated P41180 signaling to different pathways . Three mutations ( G(670)R , P(748)R , and L(773)R ) altered the binding affinity of allosteric modulators to the P41180 , and 3 mutations ( V(817)I , L(773)R , and E(767)K ) altered the cooperativity between the allosteric modulator and Ca(2+)(o) . These findings have important implications for the treatment of diseases associated with P41180 mutations using allosteric P41180 modulators and for analyzing the effects of mutations on the function and pharmacoregulation of the P41180 . Behind the curtain : cellular mechanisms for allosteric modulation of calcium-sensing receptors . DB01373 -sensing receptors ( P41180 ) are integral to regulation of systemic Ca(2+) homeostasis . Altered expression levels or mutations in P41180 cause Ca(2+) handling diseases . P41180 is regulated by both endogenous allosteric modulators and allosteric drugs , including the first Food and Drug Administration-approved allosteric agonist , DB01012 HCl ( Sensipar® ) . Recent studies suggest that allosteric modulators not only alter function of plasma membrane-localized P41180 , but regulate P41180 stability at the endoplasmic reticulum . This brief review summarizes our current understanding of the role of membrane-permeant allosteric agonists in cotranslational stabilization of P41180 , and highlights additional , indirect , signalling-dependent role(s) for membrane-impermeant allosteric drugs . Overall , these studies suggest that allosteric drugs act at multiple cellular organelles to control receptor abundance and hence function , and that drug hydrophobicity can bias the relative contributions of plasma membrane and intracellular organelles to P41180 abundance and signalling . Clinical utilization of cinacalcet in hypercalcemic conditions . INTRODUCTION : DB01012 has recently been introduced as a treatment for secondary hyperparathyroidism in dialysis patients and for parathyroid carcinoma . However , there has been an increasing interest in finding out whether cinacalcet can be used as a treatment for other parathyroid hormone ( PTH ) -dependent hypercalcemic conditions also . AREAS COVERED : The article reports the most relevant recent contributions dealing with calcium sensing receptor ( P41180 ) physiology as well as cinacalcet pharmacokinetics and pharmacodynamics . It also looks at the different hypercalcemic conditions where the use of cinacalcet has been proposed . This article was researched using clinical trials , case reports and outstanding basic research published in the last 3 years ( MEDLINE database up to 31 November 2010 ) . It provides the reader with an insight into the many unaddressed issues regarding cinacalcet that need to be resolved before it can be used in newly proposed fields . EXPERT OPINION : Since cinacalcet may not only have an effect on parathyroid P41180 but also on P41180 expressed at bone and renal levels , it can currently only be considered a good alternative to parathyroidectomy in PTH-dependent hypercalcemic conditions when surgical intervention is burdened by a high failure rate or when it can be considered a risky procedure . At present , cinacalcet can not be considered the first choice treatment in asymptomatic primary hyperparathyroidism or in mild-to-moderate forms of familial hypocalciuric hypocalcemia . Identification of a novel extracellular cation-sensing G-protein-coupled receptor . The C family G-protein-coupled receptors contain members that sense amino acid and extracellular cations , of which calcium-sensing receptor ( P41180 ) is the prototypic extracellular calcium-sensing receptor . Some cells , such as osteoblasts in bone , retain responsiveness to extracellular calcium in P41180 -deficient mice , consistent with the existence of another calcium-sensing receptor . We examined the calcium-sensing properties of Q5T6X5 , a newly identified member of this family . Alignment of Q5T6X5 with P41180 revealed conservation of both calcium and calcimimetic binding sites . In addition , calcium , magnesium , strontium , aluminum , gadolinium , and the calcimimetic P0C0P6 568 resulted in a dose-dependent stimulation of Q5T6X5 overexpressed in human embryonic kidney cells 293 cells . Also , osteocalcin , a calcium-binding protein highly expressed in bone , dose-dependently stimulated Q5T6X5 activity in the presence of calcium but inhibited the calcium-dependent activation of P41180 . Coexpression of beta-arrestins 1 and 2 , regulators of G-protein signaling P41220 or P49798 , the RhoA inhibitor P01024 toxin , the dominant negative Galpha(q)-(305-359) minigene , and pretreatment with pertussis toxin inhibited activation of Q5T6X5 by extracellular cations . Reverse transcription-PCR analyses showed that mouse Q5T6X5 is widely expressed in mouse tissues , including bone , calvaria , and the osteoblastic cell line MC3T3-E1 . These data suggest that in addition to sensing amino acids , Q5T6X5 is a cation- , calcimimetic- , and osteocalcin-sensing receptor and a candidate for mediating extracellular calcium-sensing responses in osteoblasts and possibly other tissues . [ DB01012 -- a new drug for the treatment of secondary hyperparathyroidism in patients with uraemia , parathyroid cancer or primary hyperparathyroidism ] . DB01012 is a new drug with antiparathyroid effects that belongs to the class of calcimimetics . It increases the sensitivity of the calcium-sensing receptor ( P41180 ) to calcium , thus inducing a decrease in plasma parathyroid ( PTH ) levels . In patients with uncontrolled secondary hyperparathyroidism due to uremia , cinacalcet has been shown to decrease the levels of PTH even in those optimally treated with calcium and 1-ahydroxylated vitamin D . DB01012 decreases plasma calcium and plasma PTH levels in patients with primary hyperparathyroidism or parathyroid cancer . DB01076 inhibits RhoC function and limits head and neck cancer metastasis . OBJECTIVE : RhoC oncogene is a well characterized marker of metastasis in a majority of invasive cancers , including HNSCC . Elevated RhoC expression has been found to be associated with distant metastasis . Statins are a class of drugs that are used to reduce cholesterol levels by inhibiting P04035 activity which in turns prevents mevalonate synthesis , which is a precursor for synthesis of cholesterol and prenylation . Interestingly , the proper function of Rho proteins depends on prenylation . Significantly , it has been reported that metastasis in human melanoma can be reduced by atorvastatin which inhibits RhoC activity by preventing its geranylgeranylation . Given that RhoC is a key oncogene involved in metastasis , we hypothesized DB01076 can reduce head and neck metastasis by inhibiting RhoC activity . METHODS : In vitro and in vivo studies were carried out to evaluate the ability of DB01076 to inhibit RhoC function and HNSCC metastasis . Cell motility , proliferation , cell invasion , and colony formation assays were performed according to the standard protocols . RESULTS : DB01076 treatment significantly reduced the active form of RhoC in vitro and diminished cell motility , invasion , proliferation and colony formation . Importantly , we observed a significant decrease in p- P27361 /2 and p- P40763 in DB01076 treated cell lines . In vivo experiments revealed inhibition of angiogenesis and lung metastases with DB01076 therapy . CONCLUSIONS : This study is the first of its kind to establish a potential role of DB01076 in head and neck cancer therapy . These findings suggest that DB01076 can be a potential low risk adjuvant therapy to minimize metastases in aggressive forms of HNSCC . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Regulation of extracellular signal-regulated kinase 1/2 influences hippocampal neuronal survival in a rat model of diabetic cerebral ischemia . Activation of extracellular signal-regulated kinase 1/2 has been demonstrated in acute brain ischemia . We hypothesized that activated extracellular signal-regulated kinase 1/2 can protect hippocampal neurons from injury in a diabetic model after cerebral ischemia/reperfusion . In this study , transient whole-brain ischemia was induced by four-vessel occlusion in normal and diabetic rats , and extracellular signal-regulated kinase 1/2 inhibitor ( U0126 ) was administered into diabetic rats 30 minutes before ischemia as a pretreatment . Results showed that the number of surviving neurons in the hippocampal P00915 region was reduced , extracellular signal-regulated kinase 1/2 phosphorylation and P12956 activity were decreased , and pro-apoptotic Bax expression was upregulated after intervention using U0126 . These findings demonstrate that inhibition of extracellular signal-regulated kinase 1/2 activity aggravated neuronal loss in the hippocampus in a diabetic rat after cerebral ischemia/reperfusion , further decreased DNA repairing ability and accelerated apoptosis in hippocampal neurons . P27361 /2 activation plays a neuroprotective role in hippocampal neurons in a diabetic rat after cerebral ischemia/reperfusion . A phase I-II study of the histone deacetylase inhibitor vorinostat plus sequential weekly paclitaxel and doxorubicin-cyclophosphamide in locally advanced breast cancer . Histone deacetylases ( HDACs ) are a family of enzymes that regulate chromatin remodeling and gene transcription . DB02546 is a panHDAC inhibitor that sensitizes breast cancer cells to taxanes and trastuzumab by suppressing Q9UBN7 and Hsp90 client proteins . Fifty-five patients with clinical stage IIA-IIIC breast cancer received 12 weekly doses of paclitaxel ( 80 mg/m(2) ) plus vorinostat ( 200-300 mg PO P55957 ) on days 1-3 of each paclitaxel dose plus trastuzumab ( for Her2/neu positive disease only ) , followed by doxorubicin/cyclophosphamide ( 60/600 mg/m(2) every 2 weeks plus pegfilgrastim ) . The primary study endpoint was pathologic complete response (pCR). pCR occurred in 13 of 24 evaluable patients with Her2-positive disease ( 54 , 95 % confidence intervals [ CI ] 35-72 % ) , which met the prespecified study endpoint . pCR occurred in 4 of 15 patients with triple negative disease ( 27 , 95 % CI 11-52 % ) and none of 12 patients with ER-positive , Her2/neu negative disease ( 0 , 95 % CI 0-24 % ) , which did not meet the prespecified endpoint . ER-positive tumors exhibited lower Ki67 and higher Hsp70 expression , and Q9UBN7 , Hsp70 , P38936 , and p27 expression were not predictive of response . DB02546 increased acetylation of Hsp90 and alpha tubulin , and reduced expression of Hsp90 client proteins and Q9UBN7 in the primary tumor . Combination of vorinostat with weekly paclitaxel plus trastuzumab followed by doxorubicin-cyclophosphamide is associated with a high pCR rate in locally advanced Her2/neu positive breast cancer . Consistent with cell line and xenograft data , vorinostat increased acetylation of Hsp90 and alpha tubulin , and decreased Hsp90 client protein and Q9UBN7 expression in human breast cancers in vivo . Cytological properties of stromal cells derived from giant cell tumor of bone ( GCTSC ) which can induce osteoclast formation of human blood monocytes without cell to cell contact . When human blood monocytes were cocultured with stromal cells derived from human giant cell tumor of bone ( GCTSC ) and a Millipore filter ( 0.4 microm ) was interposed between monocytes and GCTSC , multinucleated giant cell formation of monocytes was induced . The multinucleated giant cells have characters as osteoclast-like cells , indicating that a soluble osteoclast-inducing factor(s) is secreted from GCTSC expressing Q9Y6Q6 , O14788 / O14788 / O14788 and P78536 mRNA . Furthermore , O00300 / O00300 inhibited GCTSC-induced osteoclastogenesis , showing that the Q9Y6Q6 - O14788 system is involved in GCTSC-induced osteoclastogenesis and that soluble form of O14788 / O14788 induces osteoclasts from monocytes . GCTSC expressed the cytokine mRNAs such as P09603 , GM- P04141 , P08700 , P05112 , P05231 , and P01579 mRNAs . None of IL-1ralpha , IL-1alpha , IL-1beta , P60568 , P05112 , P22301 , Q14116 , P01375 , G- P04141 and P01579 could be detected in all culture media . A significant amount of P05231 could be detected in the culture media of all GCTSC . P10145 was found in the culture media of two GCTSC and two osteosarcoma-derived cells . P09603 was detected in all culture media . GCTSC express P41180 , and stimulation of GCTSC with either extracellular Ca(2+) or neomycin , agonist of P41180 , augmented the expression of O14788 . Some lines of GCTSC expressed alkaline phosphatase , osteocalcin and Cbfa1 , suggesting that GCTSC are intimately related to osteoblastic lineage . DB01373 sensing receptor modulation for cancer therapy . The calcium sensing receptor ( P41180 ) is a member of the largest family of cell surface receptors , the G protein-coupled receptors involved in calcium homeostasis . The role of the P41180 in neoplasia appears to be homeostatic ; loss of normal P41180 -induced response to extracellular calcium is observed in cancers of the colon and ovary , while increased release of P12272 is observed in cancers of the breast , prostate and Leydig cells . Currently P41180 can be considered as a molecule that can either promote or prevent tumor growth depending on the type of cancer . Therefore , recognition of the multifaceted role of P41180 in gliomas and other malignant tumors in general is fundamental to elucidating the mechanisms of tumor progression and the development of novel therapeutic agents . Emphasis should be placed on development of drug-targeting methods to modulate P41180 activity in cancer cells . Overexpression of SnoN/SkiL , amplified at the 3q26.2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26.2 encompassing SnoN/SkiL , a coregulator of SMAD/TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 promoters with little effect on the P38936 promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . P62158 regulates Ca2+-sensing receptor-mediated Ca2+ signaling and its cell surface expression . The Ca(2+)-sensing receptor ( P41180 ) is a member of family C of the GPCRs responsible for sensing extracellular Ca(2+) ( [Ca(2+)](o) ) levels , maintaining extracellular Ca(2+) homeostasis , and transducing Ca(2+) signaling from the extracellular milieu to the intracellular environment . In the present study , we have demonstrated a Ca(2+)-dependent , stoichiometric interaction between P62158 and a P62158 -binding domain ( CaMBD ) located within the C terminus of P41180 ( residues 871-898 ) . Our studies suggest a wrapping around 1-14-like mode of interaction that involves global conformational changes in both lobes of P62158 with concomitant formation of a helical structure in the CaMBD . More importantly , the Ca(2+)-dependent association between P62158 and the C terminus of P41180 is critical for maintaining proper responsiveness of intracellular Ca(2+) responses to changes in extracellular Ca(2+) and regulating cell surface expression of the receptor . P04035 inhibitors up-regulate anti-aging klotho mRNA via RhoA inactivation in IMCD3 cells . OBJECTIVE : Q9UEF7 is thought to play a critical role in the development of age-related disorders including arteriosclerosis . Statins may exert vascular protective effects , independent of the lowering of plasma cholesterol levels . We investigated the impact of statins on mRNA expression of the age-suppressor gene , klotho in mIMCD3 cells . METHODS AND RESULTS : Q9UEF7 mRNA levels were evaluated with real-time RT-PCR . DB01076 and pitavastatin increased the expression of klotho mRNA in a dose-dependent manner . This stimulatory effect was abolished by the addition of mevalonate , GGPP and FPP , essential molecules for isoprenylation of the small GTPase Rho . As was the case with the statin treatment , inhibition of Rho-kinase by Y27632 up-regulated klotho mRNA . In contrast to the statin treatment , stimulation with angiotensin II down-regulated klotho mRNA expression without obvious morphological changes . Furthermore , pretreatment with atorvastatin blunted the angiotensin II-induced response and ameliorated the decrease in klotho mRNA expression towards basal levels . RhoA activity was further evaluated by detection of its translocation . Angiotensin II activated RhoA , whereas statins potently inactivated RhoA and blocked RhoA activation by angiotensin II . CONCLUSION : Statins inactivate the RhoA pathway , resulting in over-expression of klotho mRNA , which may contribute to the novel pleiotropic effects of statins towards vascular protection . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . DB01012 HCl prevents development of parathyroid gland hyperplasia and reverses established parathyroid gland hyperplasia in a rodent model of CKD . BACKGROUND : Secondary hyperparathyroidism ( sHPT ) represents an adaptive response to progressively impaired control of calcium , phosphorus and vitamin D in chronic kidney disease ( CKD ) . It is characterized by parathyroid hyperplasia and excessive synthesis and secretion of parathyroid hormone ( PTH ) . Parathyroid hyperplasia in uremic rats can be prevented by calcium-sensing receptor ( P41180 ) activation with the calcimimetic cinacalcet ( Sensipar®/Mimpara® ) ; however , it is unknown , how long the effects of cinacalcet persist after withdrawal of treatment or if cinacalcet is efficacious in uremic rats with established sHPT . METHODS : We sought to determine the effect of cinacalcet discontinuation in uremic rats and whether cinacalcet was capable of influencing parathyroid hyperplasia in animals with established sHPT . RESULTS : Discontinuation of cinacalcet resulted in reversal of the beneficial effects on serum PTH and parathyroid hyperplasia . In rats with established sHPT , cinacalcet decreased serum PTH and mediated regression of parathyroid hyperplasia . The cinacalcet-mediated decrease in parathyroid gland size was accompanied by increased expression of the cyclin-dependent kinase inhibitor P38936 . Prevention of cellular proliferation with cinacalcet occurred despite increased serum phosphorus and decreased serum calcium . CONCLUSIONS : The animal data provided suggest established parathyroid hyperplasia can be reversed by modulating P41180 activity with cinacalcet and that continued treatment may be necessary to maintain reductions in PTH . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Expression of ras proto-oncogenes in the Dunning R3327 rat prostatic adenocarcinoma system . Steady-state levels of c-Ha-ras mRNA were measured in eight sublines of the Dunning R3327 rat prostatic adenocarcinoma . As a control , normal dorsal prostate tissue was studied . Increased expression of c-Ha-ras is associated with tumor progression in one lineage of the Dunning R3327 system ( H to AT1 to P24752 -Lu and P24752 -Ly-Lu ) . Here ras mRNA increases as the tumor advances from androgen dependence and a high degree of differentiation to an anaplastic aneuploid phenotype with high metastatic potential . However , in the other Dunning lineage ( H to HI to HI-F to P01008 ) , expression of c-Ha-ras is variable and does not correlate with tumor progression . Immunocytochemistry showed that levels of the c-Ha-ras P38936 protein paralleled steady-state mRNA levels in variants . Transfection assays , using NIH/3T3 cells , suggested that the ras loci were not activated in the R3327 tumors . Levels of P01116 mRNA were also measured in the Dunning tumors ; these did not correlate with tumor progression in either lineage . Expression of N-ras mRNA was not detected in the Dunning tumors . Discovery of a calcimimetic with differential effects on parathyroid hormone and calcitonin secretion . Calcimimetics are positive allosteric modulators to the calcium-sensing receptor ( P41180 ) . Activation of the P41180 inhibits the secretion of parathyroid hormone ( PTH ) , stimulates the secretion of calcitonin , and decreases serum calcium ( Ca(2+) ) . DB01012 , a second-generation calcimimetic , is used therapeutically to control PTH in patients with chronic kidney disease who are on dialysis with secondary hyperparathyroidism . A calcimimetic that displays increased separation of PTH versus Ca(2+) lowering in patients would potentially allow the use of calcimimetics to treat patients in earlier stages of renal disease because hypocalcemia can develop in this population . Toward this end , we developed a third-generation calcimimetic , determined the molecular pharmacological properties of it using an operation model of allosteric modulation/agonism , and measured the compound effects on PTH , serum ionized Ca(2+) , and calcitonin levels in 5/6 nephrectomized rats . We found the new molecule effectively reduced PTH levels without promoting calcitonin secretion or hypocalcemia . Furthermore , our third-generation molecule was less efficacious at promoting calcitonin secretion from human thyroid carcinoma cells compared with 3-(2-chlorophenyl)-N- ( ( 1R ) -1-(3-methoxyphenyl)ethyl ) -1-propanamine ( R-568 ) , a first-generation calcimimetic . These data provide evidence that calcimimetics with increased potency can be used to lower PTH without production of significant hypocalcemia because the threshold for inhibition of PTH secretion is much lower than the threshold for calcitonin secretion . [ Medical treatment of primary hyperparathyroidism : role of calcimimetics ] . Primary hyperparathyroidism ( PHPT ) is a common endocrinological process , characterized by chronic elevation of serum concentrations of calcium and parathyroid hormone ( PTH ) . The only intervention able to cure the disease is parathyroidectomy . However , there are few valid medical alternatives for patients whose PHPT is unresolved by surgery , or in those with contraindications for surgery or who refuse the procedure . The discovery of calcium-sensing receptors ( CaSRs ) , which regulate PTH secretion according to extracellular calcium concentrations , has allowed specific anti-parathyroid drugs called calcimimetics to be designed . DB01012 is an allosteric modulator of P41180 that has demonstrated safety and efficacy in controlling serum calcium values and in reducing PTH levels in patients with PHPT . The exact role of calcimimetics in the overall management of PHPT is promising and should be considered in future clinical practice guidelines . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . DB01012 : pharmacological and clinical aspects . The calcium sensing receptor ( P41180 ) is expressed in cells secreting calcium-regulating hormones , in cells involved in calcium transport and in many other tissues , with an as yet not completely defined role . In parathyroid cells , the P41180 stimulation inhibits parathyroid hormone ( PTH ) secretion , synthesis and parathyroid cell proliferation . DB01012 belongs to calcimimetic type II compounds that can interact with P41180 , increasing its affinity for calcium . Clinical studies have proved cinacalcet to be effective in reducing calcium and PTH levels in primary hyperparathyroidism and in reducing PTH , calcium and phosphate in patients with secondary hyperparathyroidism owing to chronic renal failure , with a relatively safe profile , the only reported adverse events being hypocalcaemia and gastrointestinal symptoms . However , though calcimimetics do represent a real advancement in the field of the treatment of PTH secretion disturbances , there is a need for clinical trials , which should aim to demonstrate that a better control of biochemical parameters is also matched with better clinical outcomes . Suppression of parathyroid hormone-related protein messenger RNA expression by medroxyprogesterone acetate in breast cancer tissues . The level of parathyroid hormone-related protein ( P12272 ) expressed in breast cancer tissue is closely related to the incidence of bone metastasis . We examined the P12272 mRNA expression in breast cancer tissues by coamplification polymerase chain reaction ( PCR ) in mole ratio to internal standard beta-actin mRNA . The P12272 expression was higher in premenopausal patients than in postmenopausal patients ( P < 0.05 ) . More pronounced difference by menopause found in estrogen receptor ( ER ) positive groups ( P < 0.001 ) indicated that the P12272 expression in breast cancer tissue is hormonally regulated and might be altered by endocrine agents . To clarify the changes of P12272 expression by endocrine therapy of breast cancer , we measured P12272 expression in the breast cancer tissue incubated for 24 h with 1 x 10(-8) M of estradiol ( E2 ) , 1 x 10(-6) M of tamoxifen ( TAM ) and 1 x 10(-5) M of medroxyprogesterone acetate ( DB00603 ) . The P12272 expression was decreased significantly by DB00603 ( P < 0.005 ) , while E2 and TAM did not change the P12272 expression . P06401 ( PgR ) mRNA expression was also examined to confirm that the breast cancer tissue responds to E2 and TAM . The results were well compatible with the better therapeutic effect of DB00603 reported for the treatment of breast cancer with bone metastases . As a potential candidate for the receptor that mediates the suppressive effect of DB00603 , androgen receptor ( AR ) is suggested most probable . Present results also demonstrated that the clinical response of individual tumors is closely associated with the early in vitro changes of gene expression detected in the cancer specimen . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . A new role for extracellular Ca2+ in gap-junction remodeling : studies in humans and rats . We wanted to elucidate whether extracellular calcium may regulate the expression of the cardiac gap-junction proteins connexin 40 and connexin43 . In the free wall of the left atria of 126 cardiac surgery patients with either sinus rhythm ( SR ) or chronic atrial fibrillation ( AF ) , we determined the expression of the cardiac gap-junction proteins P17302 and P36382 by Western blot and immunohistology . For deeper investigation , we incubated cultured neonatal rat cardiomyocytes at 2 or 4 mM Ca(++) for 24 h and determined intercellular coupling , P36382 , P17302 protein and mRNA expression , protein trafficking and sensitivity to verapamil ( 10-100 nM ) , cyclosporin A ( 1 microM ) ,and BMS605401 ( 100 nM ) , a specific inhibitor of Ca(2+)-sensing receptor ( P41180 ) . We found in patients that both Cx are up-regulated in AF in the left atrium ( by 100-200 % ) . Interestingly , P36382 was mainly up-regulated , if total serum calcium was > or=2.2 mM , while P17302 was independent from extracellular [ Ca(++) ] . In cultured cells , 4 mM Ca(++)-exposure lead to up-regulation of P36382 , but not P17302 . We found enhanced P36382 in the plasma membrane and reduced P36382 in the Golgi apparatus . The membrane P36382 up-regulation resulted in enhanced gap-junction intercellular coupling with a shift in the Boltzmann fit of voltage-dependent inactivation indicating a higher contribution of P36382 as revealed by dual whole cell voltage clamp experiments . BMS605401 could prevent all Ca(2+)-induced changes . Moreover , cyclosporin A completely abolished the Ca(2+)-induced changes , while verapamil was ineffective . We conclude that extracellular calcium ( 24 h exposure ) seems to up-regulate P36382 but not P17302 . Tp53-associated growth arrest and DNA damage repair gene expression is attenuated in mammary epithelial cells of rats fed whey proteins . Dietary protection from mammary cancer is likely coordinated through multiple signaling pathways , based on the known heterogeneity of the disease and the distinct origins of mammary tumor cells . The present study examined the modulatory effects of dietary intake of whey protein hydrolysate ( WPH ) relative to casein ( CAS ) , on mammary epithelial cell resistance to endogenous DNA damage using Tp53 gene expression and signaling as a read-out , and on systemic proapoptotic and immune surveillance activity , in young adult female Sprague-Dawley rats . Rats were fed AIN-93G diets made with CAS or WPH as the sole protein source beginning at gestation d 4 . At postnatal day ( P01160 ) 50 , mammary glands of rats fed WPH had lower levels of activated Tp53 and p38 mitogen-activated protein kinase proteins , and reduced transcript levels for Tp53-associated DNA damage repair , growth arrest , and proapoptotic genes than those of CAS-fed rats . Serum from WPH-fed rats had greater apoptotic activity in MCF-7 tumor cells than that from rats fed CAS . Serum levels of monocyte chemoattractant protein ( MCP ) -1 were higher in WPH- than in CAS-fed rats . MCF-7 cells treated with CAS serum + recombinant rat P13500 had apoptotic activity and Tp53 and P38936 gene expression levels comparable to those treated with WPH serum or recombinant P13500 . Results indicate that mammary glands of rats fed a WPH diet are more protected from endogenous DNA damage than are those of CAS-fed rats , and identify P13500 as a potential serum biomarker for the positive effects of healthy diets . DB01012 upregulates calcium-sensing receptors of parathyroid glands in hemodialysis patients . BACKGROUND : DB01012 hydrochloride ( cinacalcet ) , a calcimimetic , has been shown to upregulate calcium-sensing receptor ( P41180 ) expression in parathyroid glands of rats with chronic renal insufficiency . However , the effect of cinacalcet on the reduced P41180 expression in human parathyroid glands remains to be elucidated . METHODS : Four normal parathyroid glands and 71 hyperplastic parathyroid glands from 18 hemodialysis patients with refractory secondary hyperparathyroidism ( SHPT ) treated with ( n = 10 ; cinacalcet group ) or without ( n = 8 ; conventional group ) cinacalcet were examined immunohistochemically with a specific antibody against P41180 . The expression level of P41180 was analyzed semiquantitatively . RESULTS : Compared with normal glands , the immunohistochemical expression of P41180 was decreased significantly in both the cinacalcet and conventional groups . In the cinacalcet group , the expression of P41180 was increased significantly compared with that in the conventional group ( 1.83 ± 0.14 vs. 0.87 ± 0.15 , p < 0.001 ) , even though the proportion of patients using vitamin D sterols and the mean administered dose of calcitriol equivalents were not significantly different between the two groups . The expression of P41180 was significantly decreased in the larger glands ( > 500 mg ) compared with that in the smaller glands ( < 500 mg ) in both groups ; furthermore , it was markedly decreased in areas of nodular hyperplasia compared with diffuse hyperplasia in the cinacalcet group . CONCLUSIONS : Our results indicate that cinacalcet upregulates the depressed expression of P41180 in hemodialysis patients with SHPT , and that insufficient expression of P41180 , especially in larger glands with advanced nodular hyperplasia , underlies the pathogenesis of SHPT in patients who are resistant to cinacalcet . Connexin36 knockout mice display increased sensitivity to pentylenetetrazol-induced seizure-like behaviors . OBJECTIVE : Large-scale synchronous firing of neurons during seizures is modulated by electrotonic coupling between neurons via gap junctions . To explore roles for connexin36 ( Q9UKL4 ) gap junctions in seizures , we examined the seizure threshold of connexin36 knockout ( Cx36KO ) mice using a pentylenetetrazol ( PTZ ) model . METHODS : Mice ( 2-3months old ) with Q9UKL4 wildtype ( WT ) or Cx36KO genotype were treated with vehicle or 10-40mg/kg of the convulsant PTZ by intraperitoneal injection . Seizure and seizure-like behaviors were scored by examination of video collected for 20min . Quantitative real-time PCR ( QPCR ) was performed to measure potential compensatory neuronal connexin ( Q8NFK1 , P35212 , P17302 and P36383 ) , pannexin ( Q96RD7 and Q96RD6 ) and gamma-aminobutyric acid type A ( GABA(A) ) receptor α1 subunit gene expression . RESULTS : Cx36KO animals exhibited considerably more severe seizures ; 40mg/kg of PTZ caused severe generalized ( ≥grade III ) seizures in 78 % of KO , but just 5 % of WT mice . A lower dose of PTZ ( 20mg/kg ) induced grade II seizure-like behaviors in 40 % KO vs. 0 % of WT animals . There was no significant difference in either connexin , pannexin or GABA(A) α1 gene expression between WT and KO animals . CONCLUSION : Increased sensitivity of Cx36KO animals to PTZ-induced seizure suggests that Q9UKL4 gap junctional communication functions as a physiological anti-convulsant mechanism , and identifies the Q9UKL4 gap junction as a potential therapeutic target in epilepsy . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Clinical utility of calcimimetics targeting the extracellular calcium-sensing receptor ( P41180 ) . Calcimimetics , which activate the extracellular calcium ( Ca(o)(2+) ) -sensing receptor in the parathyroid and other tissues participating in Ca(o)(2+) homeostasis , were the first described allosteric activators of a G-protein-coupled receptor . DB01012 , the only calcimimetic currently approved for human use , is used clinically for treating secondary hyperparathyroidism ( e.g. , overactivity of parathyroid glands ) in patients being dialyzed for chronic kidney disease . By sensitizing the parathyroids to Ca(o)(2+) , cinacalcet lowers the circulating parathyroid hormone ( PTH ) level . It also reduces serum calcium and phosphate , changes increasing the percentage of patients achieving the guidelines recommended by the National Kidney Foundation ( NKF ) for these minerals . Studies are underway addressing whether better adherence to these guidelines in patients receiving cinacalcet reduces cardiovascular disease and related mortality , which are both common is the dialysis population . The second approved use of cinacalcet is for treating hypercalcemia in patients with inoperable parathyroid carcinoma . In this setting , it provides the first medical therapy chronically lowering serum calcium concentration in this condition , albeit not to normal in most patients . Its effect on the long-term prognosis of these patients , if any , is presently unclear . " Off-label " administration of cinacalcet [ i.e. , not yet approved by the US Food and Drug Administration ( FDA ) ] effectively lowers serum calcium and/or PTH in various other forms of hyperparathyroidism and increases serum phosphate in renal phosphate-wasting syndromes by reducing PTH-induced phosphaturia . In the future , the drug could conceivably be utilized to modulate the activity of the P41180 in other tissues ( i.e. , kidney , colon ) in therapeutically desirable ways . Correlates of nurse practitioners ' diagnostic and intervention performance for domestic violence . The purposes of this research were to identify diagnosis and intervention performance accuracy , variables that influence this performance accuracy , and barriers that impede performance accuracy of adult nurse practitioners ( P01160 ) and family nurse practitioners ( FNP ) for domestic violence . Two measures were developed : the Nurse Practitioner Survey ( P0C0P6 ) and the Nurse Practitioner Performance Tool . A total of 118 ANPs and FNPs completed and returned mailed surveys . Of these , 22 individuals were interviewed by telephone regarding personal and professional experience with domestic violence and barriers in their clinical settings to addressing domestic violence . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . DB02901 interacts with P00533 /MAPK signalling and modulates P00533 levels in androgen receptor-positive LNCaP prostate cancer cells . P10275 ( AR ) signalling plays a pivotal role in prostate cancer pathogenesis and progression . However , androgen-mediated AR signalling is yet to be fully understood . P00533 and Q96HU1 kinase signalling pathways play predominant roles in AR function . Therefore , we investigated the interaction of P00533 signalling and AR activity in AR-positive LNCaP cells . We found that 5alpha-dihydrotestosterone ( DB02901 ) and P01133 had a synergistic effect on AR activity as detected by a luciferase reporter system , although P01133 alone did not activate AR . Both P27361 /2 and p38 were involved in DB02901 and DB02901 / P01133 -induced AR activation as detected by specific MEK and p38 inhibitors . Furthermore , 24-h treatment of the cells with DB02901 resulted in ubiquitination and down-regulation of the P00533 . This effect could be inhibited by the anti-androgen flutamide , suggesting an androgen-dependent mechanism . On the other hand , DB02901 -treatment strongly increased AR levels in LNCaP cells . These data suggest a complex regulatory loop between activated AR and P00533 . In conclusion , activation of AR by both DB02901 and P01133 / DB02901 involves the Q96HU1 kinase pathway . Long-term activation of AR results in increase of AR levels , which through so far unknown regulatory mechanisms results in ubiquitination and degradation of the P00533 . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . DB00175 -induced proangiogenic effects depend upon extracellular P09038 . The P04035 inhibitors ( statins ) have been shown to exert several protective effects on the vasculature that are unrelated to changes in the cholesterol profile , and to induce angiogenesis . The proangiogenic effect exerted by statins has been attributed to the activation of the PI3K/Akt pathway in endothelial cells ; however , it is unclear how statins activate this pathway . DB00175 -mediated activation of Akt and MAPK occurs rapidly ( within 10 min. ) and at low doses ( 10 nM ) . Here , we hypothesized that P09038 contributes to the proangiogenic effect of statins . We found that pravastatin , a hydrophilic statin , induced phosphorylation of the FGF receptor ( FGFR ) in human umbilical vein endothelial cells . SU5402 , an inhibitor of FGFR , abolished pravastatin-induced PI3K/Akt and MAPK activity . Likewise , anti- P09038 function-blocking antibodies inhibited Akt and MAPK activity . Moreover , depletion of extracellular P09038 by heparin prevented pravastatin-induced phosphorylation of Akt and MAPK . Treatment with P09038 antibody inhibited pravastatin-enhanced endothelial cell proliferation , migration and tube formation . These observations indicate that pravastatin exerts proangiogenic effects in endothelial cells depending upon the extracellular P09038 . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Calcimimetic and calcilytic drugs for treating bone and mineral-related disorders . The calcium-sensing receptor ( P41180 ) plays a pivotal role in regulating systemic Ca(2+) homeostasis and is a target for drugs designed to treat certain disorders of bone and mineral metabolism . Calcimimetics are agonists or positive allosteric modulators of the P41180 ; they inhibit parathyroid hormone ( PTH ) secretion and stimulate renal Ca(2+) excretion . The first calcimimetic drug is cinacalcet , a positive allosteric modulator of the P41180 that is approved for treating secondary hyperparathyroidism ( Q9HD23 ) in patients on renal replacement therapy and for some forms of primary Q9HD23 characterized by clinically significant hypercalcemia . DB01012 is also being investigated as a therapy for other hypercalcemic conditions and certain hypophosphatemic disorders . Calcilytics are P41180 inhibitors that stimulate the secretion of PTH and decrease renal excretion of Ca(2+) . Although calcilytics have failed thus far as anabolic therapies for osteoporosis , they are currently being evaluated as novel therapies for new indications involving hypocalcemia and/or hypercalciuria . Successful use of bisphosphonate and calcimimetic in neonatal severe primary hyperparathyroidism . Neonatal primary hyperparathyroidism ( NPHT ) is associated with an inactivating homozygous mutation of the calcium sensing receptor ( P41180 ) . The P41180 is expressed most abundantly in the parathyroid glands and the kidney and regulates calcium homeostasis through its ability to modulate parathormone secretion and renal calcium reabsorption . NPHT leads to life threatening hypercalcemia , nephrocalcinosis , bone demineralization , and neurologic disabilities . Surgery is the treatment of choice . While waiting for surgery , bisphosphonates offer a good alternative to deal with hypercalcemia . DB01012 is a class II calcimimetic that increases P41180 affinity for calcium , leading to parathormone suppression and increased calcium renal excretion . At present , there is little evidence as to whether cinacalcet could improve the function of mutant P41180 in NPHT . We report a case of NPHT , treated successfully with bisphosphonates and cinacalcet after surgery failure . To our knowledge , it is the first time cinacalcet has been used for NPHT . DB02546 and bortezomib synergistically cause ubiquitinated protein accumulation in prostate cancer cells . PURPOSE : Protein ubiquitination is a novel strategy used to treat malignancies . We investigated whether the histone deacetylase inhibitor vorinostat ( Cayman Chemical , Ann Arbor , Michigan ) and the proteasome inhibitor bortezomib ( LC Laboratories , Woburn , Massachusetts ) would synergistically cause the accumulation of ubiquitinated proteins in prostate cancer cells . MATERIALS AND METHODS : LNCaP , PC-3 and DU 145 cells ( ATCC™ ) were treated with vorinostat and/or bortezomib . Cell viability and induction of apoptosis were assessed . In vivo efficacy was evaluated in a murine subcutaneous tumor model using PC-3 cells . The influence of androgen receptor expression on bortezomib efficacy was examined using RNA interference . Changes in the expression of ubiquitinated proteins , cell cycle associated proteins and acetylated histone were evaluated . RESULTS : P10275 expression seemed to decrease bortezomib activity . PC-3 and DU 145 cells were more susceptible to bortezomib than LNCaP cells and the silencing of androgen receptor expression in LNCaP cells enhanced bortezomib activity . DB02546 and bortezomib synergistically induced apoptosis , inhibited prostate cancer cell growth and suppressed tumor growth in a murine xenograft model . The combination decreased cyclin D1 and cyclin-dependent kinase 4 expression , and increased P38936 expression . The combination synergistically caused the accumulation of ubiquitinated proteins and histone acetylation . This histone acetylation was a consequence of the accumulation of ubiquitinated proteins . CONCLUSIONS : DB02546 and bortezomib inhibit the growth of prostate cancer cells synergistically by causing ubiquitinated proteins to accumulate in cells . The current study provides a framework for testing the combination in patients with advanced prostate cancer . Inhibition of HCV by the serpin antithrombin III . BACKGROUND : Although there have been dramatic strides made recently in the treatment of chronic hepatitis C virus infection , interferon-α based therapy remains challenging for certain populations , including those with unfavorable Q8IZI9 genotypes , psychiatric co-morbidity , HIV co-infection , and decompensated liver disease . We have recently shown that P01008 , a serine protease inhibitor ( serpin ) , has broad antiviral properties . RESULTS : We now show that P01008 is capable of inhibiting HCV in the OR6 replicon model at micromolar concentrations . At a mechanistic level using gene-expression arrays , we found that P01008 treatment down-regulated multiple host cell signal transduction factors involved in the pathogenesis of cirrhosis and hepatocellular carcinoma , including Jun , Myc and P12643 . Using a protein interactive network analysis we found that changes in gene-expression caused by P01008 were dependent on three nodes previously implicated in HCV disease progression or HCV replication : NFκB , O75791 MAPK , and P27361 /2 . CONCLUSIONS : Our findings suggest that P01008 stimulates a novel innate antiviral host cell defense different from current treatment options . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . [ The effect of blood pressure-reducing therapy with captopril on tubular marker excretion in type-1 diabetics with nephropathy ] . A prospective open clinical trial was carried out with 23 hypertensive type I diabetics ( 13 men , ten women , mean age 49 +/- 9.1 years , duration of diabetes 18 +/- 9.1 years ) with early nephropathy . Glomerular and tubular renal function and metabolic parameters were monitored during 8 months ' treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , captopril , in addition to previous antihypertensive treatment with one or more drugs . Blood pressure control tended to improve on captopril ( systolic pressures 152 +/- 13 vs 140 +/- 13 mm Hg , P < 0.05 ; diastolic pressures 89 +/- 10 vs 87 +/- 10 mm Hg , not significant ) . Proteinuria ( > 0.5 g/24 hours ) fell into the microalbuminuria range ( albumin excretion 2-20 mg/mmol creatinine ) in four out of 13 patients , and microalbuminuria disappeared in four out of ten patients . Urinary levels of the brush border enzyme O60502 ( NAG ) , a marker of tubular dysfunction , were initially raised and fell significantly after 8 months ' treatment with captopril ( 20.3 +/- 14.4 vs 8.8 +/- 8.1 U/g creatinine ; P < 0.01 ) . DB01197 did not affect metabolic control ( HbA1 , total , HDL and LDL cholesterol , triglycerides , apolipoproteins A1 and B ) or the insulin dosage . These results show that long-term treatment with captopril may favourably influence both albumin excretion and NAG activity , a marker of tubular dysfunction , in type I diabetics with nephropathy . [ Pathophysiology of secondary hyperparathyroidism : the role of Q9GZV9 and Q9UEF7 ] . Secondary hyperparathyroidism is a complex metabolic alteration secondary to chronic kidney disease ( CKD ) . Reduction of 1,25(OH)2D3 synthesis is the first derangement , followed by an increase in PTH , and , lastly , calcium and phosphate modifications . Vitamin D is a hormone whose actions take place through a specific receptor , the vitamin D receptor ( P11473 ) , which is ubiquitous . Accordingly , heterogeneous biological effects can be added to the classical effects on mineral bone metabolism . In the pathophysiology of secondary hyperparathyroidism , an important role is also played by alterations of calcium transport , which is under the control of two receptors : P11473 and P41180 ( calcium-sensing receptor ) . The expression of these receptors is reduced during CKD . Recent findings have allowed to identify a new hormonal system , the Q9GZV9 - Q9UEF7 axis , that integrates the old and simple , but now inadequate , PTH-Vit D axis . Q9GZV9 is a circulating factor produced by osteocytes that inhibits renal phosphate reabsorption and 1-alpha-hydroxylase activity . As such , Q9GZV9 is involved in phosphate homeostasis and its serum levels increase along with the progression of CKD . Interestingly , Q9GZV9 has very low affinity for its receptor and requires the activity of Q9UEF7 , an anti-aging gene , to become active . These new actors allow us to identify a bone-kidney axis , whose real physiological importance is still under evaluation . Novel strategies in drug discovery of the calcium-sensing receptor based on biased signaling . A hallmark of chronic kidney disease is hyperphosphatemia due to renal phosphate retention . Prolonged parathyroid gland exposure to hyperphosphatemia leads to secondary hyperparathyroidism characterized by hyperplasia of the glands and excessive secretion of parathyroid hormone ( PTH ) , which causes renal osteodystrophy . PTH secretion from the parathyroid glands is controlled by the calcium-sensing receptor ( P41180 ) that senses extracellular calcium . High extracellular calcium activates the P41180 causing inhibition of PTH secretion through multiple signaling pathways . DB01012 is the first drug targeting the P41180 and can be used to effectively control and reduce PTH secretion in PTH-related diseases . DB01012 is a positive allosteric modulator of the P41180 and affects PTH secretion from parathyroid glands by shifting the calcium-PTH concentration-response curve to the left . One major disadvantage of cinacalcet is its hypocalcemic side effect , which may be caused by increased P41180 -mediated calcitonin secretion from the thyroid gland . However , multiple studies indicate that PTH and calcitonin secretion are stimulated by different signaling pathways , and therefore it might be possible to develop a P41180 activating drug that selectively activates signaling pathways that inhibit PTH secretion while having no effect on signaling pathways involved in calcitonin secretion . Such a drug would have the same therapeutic value as cinacalcet in lowering PTH secretion while eliminating the side effect of hypocalcemia by virtue of it not affecting calcitonin secretion . The present review will focus on recent advancements in understanding signaling and biased signaling of the P41180 , and how that may be utilized to discover new and smarter drugs targeting the P41180 . A novel mechanism of autophagic cell death in dystrophic muscle regulated by Q99572 receptor large-pore formation and HSP90 . Q99572 is an DB00171 -gated ion channel , which can also exhibit an open state with a considerably wider permeation . However , the functional significance of the movement of molecules through the large pore ( LP ) and the intracellular signaling events involved are not known . Here , analyzing the consequences of Q99572 activation in primary myoblasts and myotubes from the Dmd(mdx) mouse model of Duchenne muscular dystrophy , we found DB00171 -induced Q99572 -dependent autophagic flux , leading to P42574 - P55210 -independent cell death . Q99572 -evoked autophagy was triggered by LP formation but not Ca(2+) influx or P28482 - P27361 phosphorylation , 2 canonical Q99572 -evoked signals . Phosphoproteomics , protein expression inference and signaling pathway prediction analysis of Q99572 signaling mediators pointed to P54652 and HSP90 proteins . Indeed , specific HSP90 inhibitors prevented LP formation , LC3-II accumulation , and cell death in myoblasts and myotubes but not in macrophages . Pharmacological blockade or genetic ablation of p2rx7 also proved protective against DB00171 -induced death of muscle cells , as did inhibition of autophagy with 3-MA . The functional significance of the Q99572 LP is one of the great unknowns of purinergic signaling . Our data demonstrate a novel outcome -- autophagy -- and show that molecules entering through the LP can be targeted to phagophores . Moreover , we show that in muscles but not in macrophages , autophagy is needed for the formation of this LP . Given that Q99572 -dependent LP and HSP90 are critically interacting in the DB00171 -evoked autophagic death of dystrophic muscles , treatments targeting this axis could be of therapeutic benefit in this debilitating and incurable form of muscular dystrophy . LPS induces cardiomyocyte injury through calcium-sensing receptor . DB01373 -sensing receptor ( P41180 ) belongs to the family C of G-protein coupled receptors . We have previously demonstrated that P41180 could induce apoptosis of cultured neonatal rat ventricular cardiomyocytes in simulated ischemia/reperfusion . It remains unknown whether the P41180 has function in lipopolysaccharide ( LPS ) -induced myocardial injure . The aim of this study was to investigate whether the P41180 plays a role in LPS-induced myocardial injury . Cultured neonatal rat cardiomyocytes were treated with LPS , with or without pretreatment with the P41180 -specific agonist gadolinium chloride ( GdCl3 ) or the P41180 -specific antagonist NPS2390 . Release of P01375 -α and P05231 from cardiomyocytes was observed . Levels of malonaldehyde ( MDA ) , lactate dehydrogenase ( LDH ) , and activity of superoxide dismutase ( SOD ) were measured . In addition , apoptosis of the cardiomyocytes , [Ca(2+)]i and level of P41180 expression were determined . The results showed that LPS increased cardiomyocytes apoptosis , [Ca(2+)]i , MDA , LDH , P01375 -α , P05231 release , and P41180 protein expression . Compared with LPS treatment alone , pretreatment with GdCl3 further increased apoptosis of cardiomyocytes , MDA , LDH , P01375 -α , P05231 release , [Ca(2+)]i , and the expression of the P41180 protein . Conversely , pretreatment with NPS2390 decreased apoptosis of cardiomyocytes , MDA , LDH , P01375 -α , P05231 release , [Ca(2+)]i and the expression of the P41180 protein . These results demonstrate that LPS could induce cardiomyocyte injury . Moreover , LPS-induced cardiomyocyte injury was related to P41180 -mediated cardiomyocytes apoptosis , P01375 -α , P05231 release , and increase of intracellular calcium . DB01373 -sensing receptor activation in chronic kidney disease : effects beyond parathyroid hormone control . Secondary hyperparathyroidism ( SHPT ) is an important complication of advanced chronic kidney disease ( CKD ) . DB01012 , an allosteric modulator of the calcium-sensing receptor ( P41180 ) expressed in parathyroid glands , is the only calcimimetic approved to treat SHPT in patients on dialysis . By enhancing P41180 sensitivity for plasma extracellular calcium ( Ca(2+)0 ) , cinacalcet reduces serum parathyroid hormone , Ca(2+)0 , and serum inorganic phosphorous concentrations , allowing better control of SHPT and CKD-mineral and bone disorders . Of interest , the P41180 also is expressed in a variety of tissues where its activation regulates diverse cellular processes , including secretion , apoptosis , and proliferation . Thus , the existence of potential off-target effects of cinacalcet can not be neglected . This review summarizes our current knowledge concerning the potential role(s) of the P41180 expressed in various tissues in CKD-related disorders , independently of parathyroid hormone control . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . Modulation of cyclin D1 expression in human tumoral parathyroid cells : effects of growth factors and calcium sensing receptor activation . The study investigated cyclin D1 regulation by growth factors and calcium sensing receptor ( P41180 ) in human tumoral parathyroid cells . Basic fibroblast and epidermal growth factors increased cyclin D1 and phosphorylated extracellular signal-regulated kinases ( pERK1/2 ) levels that were both efficiently inhibited by P41180 agonists . By contrast , in growth factors-free medium cyclin D1 levels were either unaffected or stimulated by P41180 activation independently from P27361 /2 pathway . Transforming growth factor beta ( TGFbeta ) reduced cyclin D1 levels in the majority of tumors , this effect being not influenced by P41180 activation and menin expression levels . In conclusion , in parathyroid tumors cyclin D1 expression was modulated by growth factors and P41180 activation . These data further support the oncogenic role of cyclin D1 , which resulted to be target for stimulation by P09038 and P01133 and inhibition by P41180 and TGFbeta signalling in the parathyroid . Purinergic receptor-mediated rapid depletion of nuclear phosphorylated Akt depends on pleckstrin homology domain leucine-rich repeat phosphatase , calcineurin , protein phosphatase 2A , and P60484 phosphatases . Akt is an important oncoprotein , and data suggest a critical role for nuclear Akt in cancer development . We have previously described a rapid ( 3-5 min ) and Q99572 -dependent depletion of nuclear phosphorylated Akt ( pAkt ) and effects on downstream targets , and here we studied mechanisms behind the pAkt depletion . We show that cholesterol-lowering drugs , statins , or extracellular DB00171 , induced a complex and coordinated response in insulin-stimulated A549 cells leading to depletion of nuclear pAkt . It involved protein/lipid phosphatases P60484 , pleckstrin homology domain leucine-rich repeat phosphatase ( O60346 and -2 ) , protein phosphatase 2A ( PP2A ) , and calcineurin . We employed immunocytology , immunoprecipitation , and proximity ligation assay techniques and show that O60346 and calcineurin translocated to the nucleus and formed complexes with Akt within 3 min . Also P60484 translocated to the nucleus and then co-localized with pAkt close to the nuclear membrane . An inhibitor of the scaffolding immunophilin FK506-binding protein 51 ( FKBP51 ) and calcineurin , FK506 , prevented depletion of nuclear pAkt . Furthermore , okadaic acid , an inhibitor of PP2A , prevented the nuclear pAkt depletion . Chemical inhibition and siRNA indicated that O60346 , PP2A , and P60484 were required for a robust depletion of nuclear pAkt , and in prostate cancer cells lacking P60484 , transfection of P60484 restored the statin-induced pAkt depletion . The activation of protein and lipid phosphatases was paralleled by a rapid proliferating cell nuclear antigen ( P12004 ) translocation to the nucleus , a P12004 - P38936 (cip1) complex formation , and cyclin D1 degradation . We conclude that these effects reflect a signaling pathway for rapid depletion of pAkt that may stop the cell cycle . Calcimimetics in the chronic kidney disease-mineral and bone disorder . Mineral and bone disorders ( MBD ) are both an early and very common complication of chronic kidney disease ( CKD ) . It is now accepted that they represent a significant risk factor , explaining the high cardiovascular morbidity and mortality in CKD patients . During the last decade , we have been witnessing many advances in the nomenclature , classification , pathophysiology , diagnosis , and treatment of CKD and some of its complications , such as CKD-MBD . The identification of the calcium-sensing receptor ( P41180 ) involvement in the pathogenesis of primary and secondary hyperparathyroidism ( SHPT ) and the availability of a new class of drugs called calcimimetics are two outstanding examples . DB01012 , the only available calcimimetic , has been shown to be a very effective therapeutic tool in CKD-MBD . Many clinical trials with cinacalcet in hemodialysis patients with SHPT have shown a reduction in parathyroid hormone , calcium ( Ca ) , phosphate ( P ) and Ca x P product levels , allowing far greater success in reaching therapeutic goals as recommended by international guidelines . Additionally , some studies have shown that the use of cinacalcet may improve other aspects of CKD-MBD , reducing the risk of vascular calcification and parathyroidectomy , among others . Prospective studies on dialysis patients , with hard endpoint data , are currently underway . This review summarizes the most significant aspects of calcimimimetics based on both experimental and clinical results , underlining their possibilities not only for the treatment of isolated SHPT but also for other CKD-MBD related conditions . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation .

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Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Urgosedin inhibits hypotension , hypoglycemia , and pro-inflammatory mediators induced by lipopolysaccharide . Urgosedin is a newly synthesized compound especially with serotonergic and alpha-adrenergic blocking actions . In rat isolated thoracic aorta , urgosedin competitively antagonized norepinephrine- , clonidine- , and serotonin-induced vasocontractions in a concentration-dependent manner . In radioligand binding experiments , urgosedin had significant binding affinities on alpha1/alpha2 , P08908 , P28222 and 5- Q13049 receptors . Intravenous injection of lipopolysaccharide ( LPS ) produced a biphasic hypotension in normotensive rats . Although intravenous injection of urgosedin caused minor depressor actions in the normotensive Wistar rat , urgosedin significantly attenuated the secondary prolonged hypotension produced by LPS . The plasma levels of cytokines ( IL-1beta , P05231 , P01375 , and P01579 ) and hypoglycemia induced by LPS were also reduced by urgosedin . Moreover , the acute survival rates ( 350 minutes ) of endotoxic shock increased from 0 % ( LPS group ) to 100 % in the groups pretreated with urgosedin . In RAW264.7 cells , urgosedin inhibited LPS-induced inducible nitric oxide synthase ( P35228 ) expression . In conclusion , our data suggest that urgosedin was a newly potent serotonergic and mild alpha-adrenergic blocking agent . Its prevention of LPS-induced hypotension and hypoglycemia might partially mediate through its inhibition activities on the P35228 expression and cytokines formation . Urgosedin might be an effective pharmacological agent against LPS-induced hypotension , hypoglycemia , and the formation of pro-inflammatory mediators . Constitutively active Gq/11-coupled receptors enable signaling by co-expressed G(i/o)-coupled receptors . Co-expression of guanine nucleotide-binding regulatory ( G ) protein-coupled receptors ( GPCRs ) , such as the G(i/o)-coupled human P28222 ( 5-HT(1B)R ) , with the G(q/11)-coupled human histamine 1 receptor ( P35367 ) results in an overall increase in agonist-independent signaling , which can be augmented by 5-HT(1B)R agonists and inhibited by a selective inverse 5-HT(1B)R agonist . Interestingly , inverse P35367 agonists inhibit constitutively P35367 -mediated as well as 5-HT(1B)R agonist-induced signaling in cells co-expressing both receptors . This phenomenon is not solely characteristic of 5-HT(1B)R ; it is also evident with muscarinic M2 and adenosine A1 receptors and is mimicked by mastoparan-7 , an activator of G(i/o) proteins , or by over-expression of Gbetagamma subunits . Likewise , expression of the G(q/11)-coupled human cytomegalovirus ( HCMV ) -encoded chemokine receptor US28 unmasks a functional coupling of G(i/o)-coupled P32246 receptors that is mediated via the constitutive activity of receptor US28 . Consequently , constitutively active G(q/11)-coupled receptors , such as the P35367 and HCMV-encoded chemokine receptor US28 , constitute a regulatory switch for signal transduction by G(i/o)-coupled receptors , which may have profound implications in understanding the role of both constitutive GPCR activity and GPCR cross-talk in physiology as well as in the observed pathophysiology upon HCMV infection . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . DB00998 : a selective type 1B/1D serotonin receptor agonist for the treatment of migraine headache . DB00998 belongs to an innovative family of compounds aimed at breaking through the long-standing barrier of migraine headache understanding and treatment . While a typology of headaches has been recognized for some time , and a number of therapies have been introduced for reduction of headache pain and duration , the causes of migraine remain a subject of debate . Those prone to attacks continue to endure them and suffer the related symptoms such as nausea and disorientation . DB00998 , like all the triptans , acts by inducing vasoconstriction of the meningeal arteries . It has been shown in pharmacological tests to act selectively as a potent agonist of serotonin P28222 /1D receptors . DB00998 has been well tolerated in humans and efficacious in reducing headache pain and duration in clinical trials , which have also indicated that dose adjustments for age or gender are not necessary for the drug . Patients have found the use of frovatriptan acceptable over the long-term , and overall a low-incidence of adverse effects has been reported . Though not a prophylactic , frovatriptan has demonstrated the potential to significantly improve the therapeutic approaches to the treatment of migraine . A review of the use of frovatriptan in the treatment of menstrually related migraine . Menstrual migraine ( MM ) is a highly prevalent condition associated with considerable disability . Migraine attacks occur exclusively around the menstrual period in approximately 10 % of women with migraine , that is , pure menstrual migraine , while at least 50 % of them also experience migraine at other times of the month , that is , menstrually related migraine ( MRM ) . The therapeutic approach to patients with MRM is based on treatment of the attack , or prophylactic strategies . Triptans are recommended as first-line treatments for moderate to severe migraine attacks , including MM . DB00998 is one of the newest triptans . Its high affinity for P28222 /1D receptors and long half-life contribute to its distinctive clinical effect , characterized by a more sustained and prolonged effect than other triptans . Indeed , frovatriptan proved to be effective in treating the acute attack , but was particularly effective in the short-term preventive therapy of MM . In addition , frovatriptan is one of the safest triptans , with the lowest risk of treatment-emergent adverse events . Following extensive evidence from randomized pharmacological trials , frovatriptan has now gained a grade A recommendation from the guidelines for short-term prophylaxis of MM . Recent post-hoc analyses of direct comparative trials also suggest that frovatriptan might have an important role in the acute treatment of MRM . In these studies , frovatriptan showed pain relief and pain-free rates similar to those of zolmitriptan , rizatriptan , and almotriptan , but with significantly lower recurrence rates . More well-designed , randomized , prospective studies , specifically enrolling women with MM , will be needed in the near future to confirm the efficacy of frovatriptan in this migraine subtype . DB00998 Vernalis . Vanguard ( now Vernalis ) has developed frovatriptan , a selective P28222 /1D partial agonist licensed from GlaxoSmithKline as a potential treatment for migraine [ 188478 ] , [ 194382 ] , [ 377863 ] . DB00998 . black triangle DB00998 , a new serotonin receptor agonist developed for the acute treatment of migraine , has high affinity for serotonin P28222 and P28221 receptor subtypes and is a potent stimulator of contraction in human basilar arteries. black triangle A long terminal elimination half-life ( approximately 26 hours ) is a distinctive pharmacokinetic feature of frovatriptan which appears to be independent of dose , age , gender and renal function. black triangle A single oral dose of frovatriptan 2.5mg was effective in the acute treatment of migraine providing meaningful relief within 2 hours to approximately twice as many recipients as placebo in clinical trials. black triangle Consistent relief of migraine symptoms was achieved in patients who treated a number of consecutive attacks with frovatriptan and the incidence of 24-hour migraine recurrence was reduced. black triangle DB00998 was well tolerated in clinical trials , with the overall incidence of adverse events occurring with frovatriptan 2.5mg only slightly higher than that reported with placebo . Mild to moderate fatigue , nausea and paraesthesia were the most commonly reported drug-related adverse events . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Biochemical markers and genetic research of ADHD . ADHD ( attention hyperactivity disorder ) is a polygenetic disorder with various candidate genes . At this time , more than thirty dopaminergic , noradrenergic , serotonergic and GABA-ergic genes are known . The research of only some candidate genes ( P21917 , Q01959 , P21918 , P09172 , P31645 , P28222 and P60880 ) brought relatively consistent results confirming the heredity of ADHD syndromes . The results of research of other genes ( P14416 , P35462 , MAO , ADR2A , GABA A3 , GABA B3 ) are not clear yet . This paper summarizes the most important genetic data in correlations with biochemical periphery parameters ( especially for P09172 , HVA , MHPG , serotonin ) . Hypothetically , certain subgroups of ADHD may be identified by correlation of biochemical characteristics and some candidate genes . The paper discusses some implications for future research. Review . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . 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 . DB00998 , a P28222 /1D receptor agonist for migraine . DB00998 is one of the most recent serotonin receptor agonists to receive FDA , approved labelling for use in the acute management of migraine with or without aura in adults . The mechanism of action of frovatriptan is thought to be similar to that of a serotonin agonist . However , frovatriptan has distinctive pharmacokinetic and pharmacologic properties , chiefly , a high affinity for serotonin receptors 1B and 1D and a long elimination half-life ; frovatriptan was shown to be more selective for cerebral than coronary arteries , a property which makes frovatriptan more favourable in patients at risk of coronary artery disease . Additionally , frovatriptan has a half-life of approximately 25 h , substantially longer than that of any other agent within its class . This property makes frovatriptan suitable for patients who typically suffer migraines of long duration and/or those who suffer migraine recurrence . The efficacy of frovatriptan in the treatment of acute migraine was demonstrated in five double-blind , randomised , placebo-controlled trials . At 2h , headache response rates for frovatriptan 2.5 mg ranged from 38 to 40 % compared to 22-35 % for placebo . Headache recurrence for frovatriptan 2.5 mg at 24h ranged from 9 to 14 % compared with 18 % in placebo subjects . DB00998 has no clinically significant pharmacokinetic interactions with drugs used for migraine prophylaxis or with commonly prescribed medications . Adverse effects of frovatriptan including dizziness , paresthesia , dry mouth , fatigue and flushing were generally mild and well tolerated . Given the fact that patient response to serotonin agonists is individualised , and selecting an effective agent may involve trial and error , frovatriptan is a welcome alternative in the acute management of migraine . The efficacy and tolerability of frovatriptan and dexketoprofen for the treatment of acute migraine attacks . DB00998 is a DB00669 characterized by a high affinity for P28222 /1D receptors and a long half-life contributing to a more sustained and prolonged action than other triptans . DB09214 is a nonsteroidal anti-inflammatory drug with a relatively short half-life and rapid onset of action , blocking the action of cyclo-oxygenase , which is involved in prostaglandins ' production , thus reducing inflammation and pain . Both drugs have been successfully employed as monotherapies for the treatment of acute migraine attacks . The combination of these two drugs ( frovatriptan 2.5 mg plus dexketoprofen 25 or 37.5 mg ) has been tested in migraine sufferers , showing a rapid and good initial efficacy , with 2-h pain free rates of 51 % , and a high persistence in the 48-h following the onset of pain : recurrence occurred in only 29 % of attacks and sustained pain free rates were 43 % at 24- and 33 % at 48-h . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . Deliberate self-harm is associated with allelic variation in the tryptophan hydroxylase gene ( P17752 A779C ) , but not with polymorphisms in five other serotonergic genes . BACKGROUND : There is a heritable component to suicidal behaviour , encouraging the search for the associated risk alleles . Given the putative role of the 5-HT ( 5-hydroxytryptamine ; serotonin ) system in suicidal behaviour , serotonergic genes are leading candidates . In particular , several studies have reported an association with variants in the tryptophan hydroxylase ( P17752 ) gene . METHOD : We studied six serotonergic gene polymorphisms in a well-characterized sample of 129 deliberate self-harm subjects and 329 comparison subjects . The polymorphisms were P17752 ( A779C ) , 5-HT transporter ( 5-HTT , LPR S/L ) , monoamine oxidase A ( P21397 G941T ) , P28222 receptor ( P28222 G861C ) , 5- Q13049 receptor ( P28223 T102C ) , and P28335 receptor ( P28335 Cys23Ser ) . Genotyping was done using polymerase chain reaction ( PCR ) -based assays . The primary analyses compared allele and genotype frequencies between cases and controls . There were a limited number of planned secondary analyses within the deliberate self-harm group . RESULTS : The P17752 A779 allele was more common in deliberate self-harm subjects than in controls ( OR 1.38 , 95 % CI 1.02-1.88 ; P = 0.03 ) . None of the other polymorphisms was associated with deliberate self-harm . Within the deliberate self-harm group there were no associations with impulsivity , suicide risk , lifetime history of depression , or family history of deliberate self-harm . CONCLUSIONS : Our data extend the evidence that allelic variation in the P17752 gene is a risk factor for deliberate self-harm . No evidence was found to implicate the other polymorphisms . DNA sequence polymorphisms in genes involved in the regulation of dopamine and serotonin metabolism in rhesus macaques . A systematic search was performed for DNA sequence variation in genes regulating neurotransmitter metabolism in rhesus macaque ( Macaca mulatta ) . These genes included dopamine and serotonin receptors and transporters , and tyrosine hydroxylase . A total of 13 single nucleotide polymorphisms in five different genes were identified , namely : P21728 ( -244T- > G ) , q = 0.45 ; P21728 ( -179C- > T ) , q = 0.19 ; P21728 ( -127G- > A ) , q = 0.25 ; P21728 ( -11T- > G ) , q = 0.08 ; P21728 ( -81C- > T ) , q = 0.19 ; P35462 ( 248G- > A ) , q= 0.08 ; P35462 ( 341G- > C ) , q = 0.11 ; P35462 ( 377A- > G ) , q = 0.19 ; P35462 ( 403C- > T ; A59V ) , q= 0.11 ; P21917 ( 2608G- > A ) , q= 0.48 ; P28221 ( -506G- > T ) , q = 0.47 ; P28221 ( -173C- > T ) , q = 0.47 ; and HTT ( 340G- > A ) , q = 0.39 . The nucleotide positions listed correspond to the human homologs . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Meta-analysis of oral DB00669 therapy for migraine : number needed to treat and relative cost to achieve relief within 2 hours . OBJECTIVE : To determine the cost-effectiveness of the P28222 /1D agonists , or triptans , in the acute treatment of migraine . METHODS : To determine the cost-effectiveness of the triptans , a meta-analysis was conducted of the efficacy data from 27 oral DB00669 trials , using the endpoint of " pain-free " status within 2 hours after initial dosing as the indicator of efficacy . Efficacy data were used to determine the number needed to treat ( Q13423 ) to achieve pain-free status in 1 patient within 2 hours postdose and then applied the per-dose costs for each DB00669 to the Q13423 values . RESULTS : Rizatriptan 10 mg and almotriptan 12.5 mg were the most cost-effective of the triptans , costing $ 48.34 and $ 48.57 US dollars , respectively , to achieve pain-free status in 1 patient within 2 hours postdose . DB00998 2.5 mg was the most costly , with a cost-effective ratio of $ 162.49 US dollars . All other triptans fell between these extremes : zolmitriptan 5 mg ( $ 65.18 US dollars ) , sumatriptan 100 mg ( $ 70.83 US dollars ) , sumatriptan 50 mg ( $ 75.67 US dollars ) , zolmitriptan 2.5 mg ( $ 78.74 US dollars ) , and naratriptan 2.5 mg ( $ 141.43 US dollars ) , in decreasing order of cost-effectiveness . CONCLUSION : Using an Q13423 analysis , the least-costly drugs to achieve migraine cure within 2 hours are rizatriptan 10 mg and almotriptan 12.5 mg . From a population health perspective , the lower acquisition cost of almotriptan 12.5 mg allows for effective treatment of more patients than rizatriptan 10 mg for no additional medication cost . Serotonergic stimulation of corticotropin-releasing hormone and pro-opiomelanocortin gene expression . The neurotransmitter serotonin ( 5-HT ) stimulates adrenocorticotropic hormone ( DB01285 ) secretion from the anterior pituitary gland via activation of central 5-HT1 and 5-HT2 receptors . The effect of 5-HT is predominantly indirect and may be mediated via release of hypothalamic corticotropin-releasing hormone ( P06850 ) . We therefore investigated the possible involvement of P06850 in the serotonergic stimulation of DB01285 secretion in male rats . Increased neuronal 5-HT content induced by systemic administration of the precursor 5-hydroxytryptophan ( 5-HTP ) in combination with the 5-HT reuptake inhibitor fluoxetine raised P06850 mRNA expression in the paraventricular nucleus ( PVN ) by 64 % , increased pro-opiomelanocortin ( P01189 ) mRNA in the anterior pituitary lobe by 17 % and stimulated DB01285 secretion five-fold . Central administration of 5-HT agonists specific to P08908 , P28222 , 5- Q13049 or P28335 receptors increased P06850 mRNA in the PVN by 15-50 % , P01189 mRNA in the anterior pituitary by 15-27 % and DB01285 secretion three- to five-fold , whereas a specific 5- Q9H205 agonist had no effect . Systemic administration of a specific anti- P06850 antiserum inhibited the DB01285 response to 5-HTP and fluoxetine and prevented the 5-HTP and fluoxetine-induced P01189 mRNA response in the anterior pituitary lobe . Central or systemic infusion of 5-HT increased DB01285 secretion seven- and eight-fold , respectively . Systemic pretreatment with the anti- P06850 antiserum reduced the DB01285 responses to 5-HT by 80 % and 64 % , respectively . It is concluded that 5-HT via activation of P08908 , 5- Q13049 , P28335 and possibly also P28222 receptors increases the synthesis of P06850 in the PVN and P01189 in the anterior pituitary lobe , which results in increased DB01285 secretion . Furthermore , the results indicate that P06850 is an important mediator of the DB01285 response to 5-HT . Dual P00533 and P42345 targeting in squamous cell carcinoma models , and development of early markers of efficacy . The epidermal growth factor receptor ( P00533 ) is a validated target in squamous cell carcinoma ( SCC ) of the head and neck . Most patients , however , do not respond or develop resistance to this agent . P42345 ( P42345 ) is involved in the pathogenesis of SCC of the head and neck ( SCCHN ) . This study aimed to determine if targeting P42345 in combination with P00533 is effective in SCC , and to develop early pharmacodynamic markers of efficacy . Two SCC cell lines , one resistant ( HEP2 ) and one of intermediate susceptibility ( Detroit 562 ) to P00533 inhibitors , were xenografted in vivo and treated with an P42345 inhibitor ( temsirolimus ) , an P00533 inhibitor ( erlotinib ) or a combination of both . DB06287 exerted superior growth arrest in both cell lines than erlotinib . The combined treatment resulted in synergistic antitumor effects in the Detroit 562 cell line . Immunohistochemical assessment of pharmacodynamic effects in fine-needle aspiration ( FNA ) biopsies early after treatment using phospho MAPK , Phospho-P70 and Ki67 as end points demonstrated pathway abrogation in the Detroit 562 tumours treated with the combination , the only group where regressions were seen . In conclusion , an P42345 inhibitor showed antitumor activity in P00533 -resistant SCC cell lines . Marked antitumor effects were associated with dual pathway inhibition , which were detected by early FNA biopsies . Estrogen regulation of uterine genes in vivo detected by complementary DNA array . INTRODUCTION : In the present study , our aim was to identify differentially expressed genes involved in estrogen actions at the endometrium level in rats . METHODS : Thirty adult rats were ovariectomized four days prior to drug administration for 48 days . Rats were divided in 2 groups : I , control and II , conjugated equine estrogens ( CCE ) . Total RNA was isolated from uterus , and differential expression was analyzed by array technology and RT-PCR . RESULTS : A total of 32 candidate genes were shown to be upregulated or downregulated in groups I or II . Among them , differential expression was already confirmed by RT-PCR for P24593 , P28222 , c-kit , and P15692 , genes whose expression was up regulated during CCE therapy , and casein kinase II and serine kinase expression was the same level in both groups . CONCLUSION : We have demonstrated that cDNA array represents a powerful approach to identify key molecules in the estrogens therapy . A number of the candidates reported here should provide new markers that may contribute to the detection of target estrogen receptor . This information may also aid the development of new approaches to therapeutic intervention . Involvement of P28222 receptors in DB00669 -induced contractile responses in guinea-pig isolated iliac artery . Using a series of triptans we characterized in vitro the 5-hydroxytryptamine ( 5-HT ) receptor that mediates the contraction in guinea-pig iliac arteries moderately precontracted by prostaglandin F2alpha ( PGF2alpha ) . Additionally , we investigated by reverse-transcriptase polymerase chain reaction ( RT-PCR ) which DB00669 -sensitive receptor is present in this tissue . DB00998 , zolmitriptan , rizatriptan , naratriptan , sumatriptan , and almotriptan contracted guinea-pig iliac arteries with pD2 values of 7.52+/-0.04 , 6.72+/-0.03 , 6.38+/-0.06 , 6.22+/-0.05 , 5.86+/-0.05 and 5.26+/-0.04 respectively . For comparison , the pD2 values for 5-HT and 5-carboxamidotryptamine ( 5-CT ) were 7.52+/-0.02 and 7.55+/-0.03 respectively . In contrast to all other triptans tested , the concentration-response curve for eletriptan was biphasic ( first phase : 0.01-3 microM , pD2 approximately 6.6 ; second phase : > or = 10 microM ) . Contractions to 5-HT , 5-CT , frovatriptan , zolmitriptan , rizatriptan , naratriptan , sumatriptan , almotriptan , and eletriptan ( first phase ) were antagonized by the P28222 /1D receptor antagonist GR127935 ( 10 nM ) and the P28222 receptor antagonist SB216641 ( 10 nM ) . RT-PCR studies in guinea-pig iliac arteries showed a strong signal for the P28222 receptor while expression of P28221 and P30939 receptors was not detected in any sample . The present results demonstrate that DB00669 -induced contraction in guinea-pig iliac arteries is mediated by the P28222 receptor . The guinea-pig iliac artery may be used as a convenient in vitro model to study the (cardio)vascular side-effect potential of anti-migraine drugs of the DB00669 family . Disabling the mitotic spindle and tumor growth by targeting a cavity-induced allosteric site of survivin . Survivin is a member of the inhibitor of apoptosis protein family and has an essential role in mitosis . Survivin is overexpressed in a large variety of human cancers and represents an attractive target for cancer therapy . P00533 and Her/neu-transformed human tumors in particular exhibit high levels of survivin . The survivin protein forms dimers through a conserved region that is critical for subcellular localization and biological functions of the protein . We identified small molecules that target a specific cavity adjacent to the survivin dimerization surfaces . P28222 , a lead compound identified in the screen , can bind to the survivin protein at the intended target site . Moreover , P28222 alters spindle formation , causing mitotic arrest and cell death , and inhibits tumor growth in vitro and in vivo . Cell death occurs in premetaphase stage following mitotic arrest and is not a consequence of general toxicity . Thus , the study validates a novel therapeutic target site in the survivin protein and provides a promising strategy to develop a new class of therapeutic small molecules for the treatment of human cancers . Differential functional activity of 5-hydroxytryptamine receptor ligands and beta adrenergic receptor antagonists at 5-hydroxytryptamine1B receptor sites in Chinese hamster lung fibroblasts and opossum renal epithelial cells . Functional activity of 5-hydroxytryptamine ( 5-HT ) receptor ligands and beta adrenergic receptor antagonists was studied at P28222 receptor sites in Chinese hamster lung ( CHL ) fibroblasts by measuring two cellular responses : inhibition of forskolin-stimulated cyclic AMP formation and potentiation of basic fibroblast growth ( BFGF ) induced mitogenesis . A good correlation was found between the potency of agonists to inhibit forskolin-induced cyclic AMP formation and their potency to potentiate P09038 -induced thymidine incorporation in CHL fibroblasts . Potent agonist activity was measured with 5-methoxy-3,1,2,3,6-tetrahydro-4-pyidinyl- 1H-indole ( RU 24,969 ) , 5-carboxamidotryptamine ( 5-CT ) , 3-(1,2,5,6)-tetrahydro-4-pyridyl-5-pyrrolo(3,2-b)pyril-5-one ( CP 93,129 ) and 5-HT , whereas sumatriptan displayed weak agonist activity at concentrations different from its binding affinity for P28222 binding sites . In contrast to the observed P28222 receptor-mediated agonist activity in opossum kidney cells for metergoline and the beta adrenergic receptor antagonists : cyanopindolol , 4-(3-tert-butyl-amino-2-hydroxypropoxy)-indole-2 carbonic acid isopropyl ester ( SDZ 21,009 ) , isamoltane , (-)-propranolol and (-)-pindolol , antagonist activity at P28222 receptor sites was yielded in CHL fibroblasts in accordance with the reported observations at rat brain P28222 receptors . Methiothepin was the only compound that antagonized both the opossum kidney cell and CHL fibroblast P28222 receptor-mediated responses although the antagonist effect was more pronounced in CHL fibroblasts. ( ABSTRACT TRUNCATED AT 250 WORDS ) Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Pharmacokinetic evaluation of frovatriptan . INTRODUCTION : Migraine is the most common painful neurological disorder , affecting 13 % of the general population . Triptans represent a powerful pharmacological tool in acute migraine treatment , however , a significant portion of treated patients can not have access to this class due to possible adverse affects . Today , a total of seven DB00669 molecules are available , representing a commonly prescribed migraine treatment . Although there is a need of extensive use of triptans , only 25 % of migraine patients are using triptans . AREAS COVERED : This review includes triptans and evidence for the use of frovatriptan . A systematic approach is used to discuss the pharmacodynamic and pharmacokinetic aspects of frovatriptan , considering the emerging data on the clinical efficacy of frovatriptan in the treatment of migraine and cluster headaches . The data were obtained by searching the following key words in MEDLINE : pharmacokinetic , pharmacodynamic , triptans , frovatriptan , migraine , menstrual migraine , relatively to the period 1988 - 2011 . EXPERT OPINION : DB00998 has been developed in order to improve safety and efficacy of triptans . It shows a favorable tolerability and efficacy profile , limited to 24/48-h headache recurrence , when compared with other triptans . Preclinical data suggest that the pharmacokinetic profile of frovatriptan may differ from other available triptans . In fact , among triptans , frovatriptan showed the highest potency at the P28222 receptor ( 8.2 ) and the longer half-life ( 26 h ) . These parameters determine the clinical properties of frovatriptan ; in particular the lowest rate of headache recurrence in comparison with other triptans . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP .

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Development of a cell-based assay for the detection of anti-aquaporin 1 antibodies in neuromyelitis optica spectrum disorders . OBJECTIVE : To develop a cell-based assay ( CBA ) to detect aquaporin 1 ( P29972 ) antibodies and determine sensitivity/specificity in patients with neuromyelitis optica ( NMO ) spectrum disorders . METHODS : A P29320 -293T transfected cell model expressing P29972 was established and detected to be serum P29972 antibodies . RESULTS : P29972 antibodies were present in 73/98 ( 74.5 % ) P55087 antibody-positive patients . Some P55087 antibody-negative patients were also P29972 antibody-positive . Test sensitivity was 74.5 % in 98 P55087 antibody-positive patients . Test specificity was 79.6 % in 67 multiple sclerosis ( MS ) patients and 31 controls . CONCLUSION : A sensitive and simple CBA was developed to detect serum P29972 antibodies . P29972 antibodies were mainly present in NMO and its high-risk syndrome , but also in some MS patients . DB00731 , a D-phenylalanine derivative lacking either a sulfonylurea or benzamido moiety , specifically inhibits pancreatic beta-cell-type K( DB00171 ) channels . A novel antidiabetic agent , nateglinide , is a D-phenylalanine derivative lacking either a sulfonylurea or benzamido moiety . We examined with the patch-clamp method the effect of nateglinide on recombinant DB00171 -sensitive K(+) ( K( DB00171 ) ) channels expressed in human embryonic kidney 293T cells transfected with a Kir6.2 subunit and either of a sulfonylurea receptor ( Q09428 ) 1 , SUR2A , and SUR2B . In inside-out patches , nateglinide reversibly inhibited the spontaneous openings of all three types of Q09428 /Kir6.2 channels . DB00731 inhibited Q09428 /Kir6.2 channels with high and low affinities ( K(i) = 75 nM and 114 microM ) but SUR2A/Kir6.2 and SUR2B/Kir6.2 channels only with low affinity ( K(i) = 105 and 111 microM , respectively ) . DB00731 inhibited the K( DB00171 ) current mediated by Kir6.2 lacking C-terminal 26 amino acids only with low affinity ( K(i) = 290 microM ) in the absence of Q09428 . Replacement of serine at position 1237 of Q09428 to tyrosine [ Q09428 (S1237Y) ] specifically abolished the high-affinity inhibition of Q09428 /Kir6.2 channels by nateglinide . MgADP or MgUDP ( 100 microM ) augmented the inhibitory effect of nateglinide on Q09428 /Kir6.2 but not Q09428 (S1237Y)/Kir6.2 or SUR2A/Kir6.2 channels . This augmenting effect of MgADP was also observed with the Q09428 /Kir6.2(K185Q) channel , which was not inhibited by MgADP , but not with the Q09428 (K1384A)/Kir6.2 channel , which was not activated by MgADP . These results indicate that therapeutic concentrations of nateglinide ( approximately 10 microM ) may selectively inhibit pancreatic type Q09428 /Kir6.2 channels through Q09428 , especially when the channel is activated by intracellular MgADP , even though the agent does not contain either a sulfonylurea or benzamido moiety . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . Distinct interaction of nilotinib and imatinib with P-Glycoprotein in intracellular accumulation and cytotoxicity in CML Cell Line K562 cells . DB04868 , a second-generation tyrosine kinase inhibitor ( TKI ) , has been approved for first-line chronic myeloid leukemia ( CML ) treatment . The improved clinical response of nilotinib over that of the first generation TKI , imatinib , has been thought to be a result of its high potency of inhibition of P11274 - P00519 kinase . This study aimed to characterize differences between nilotinib and imatinib in the intracellular accumulation and cytotoxic effect on the CML cell line K562 . Accumulation of nilotinib in K562 cells was from 4.7- to 9.0-fold higher than that of imatinib . The cytotoxic effect of nilotinib on K562 cells was 14.2-fold higher than that of imatinib . Inhibition experiments in K562 cells , and examination of the cellular uptake using influx transporter-transfected human embryonic kidney ( P29320 ) 293 cells , suggested that the influx transporters OCT1 and P46721 , which have been reported to mediate accumulation of imatinib in CML cells , contributed little to the uptake of nilotinib . DB04868 was found to accumulate in imatinib-resistant K562 ( K562/IM ) cells overexpressing the efflux transporter P-glycoprotein ( P-gp ) , although cytotoxic assays showed that K562/IM cells displayed 20000-fold greater resistance to nilotinib over the parent K562 cells . In conclusion , the present findings suggest that intracellular accumulation of nilotinib in CML cells contributes to its clinical response and efficacy in CML patients . Although nilotinib has been reported to be effective against imatinib-resistant P00519 kinase mutants , the drug could not overcome imatinib resistance acquired by P-gp-overexpression . These results imply that classification of mechanisms of drug resistance is important for suitable strategies to treat imatinib-resistant CML patients . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . Inhibition of O60603 signaling by small molecule inhibitors targeting a pocket within the O60603 TIR domain . Toll-like receptor ( TLR ) signaling is initiated by dimerization of intracellular Toll/IL-1 receptor resistance ( TIR ) domains . For all TLRs except O15455 , recruitment of the adapter , myeloid differentiation primary response gene 88 ( MyD88 ) , to TLR TIR domains results in downstream signaling culminating in proinflammatory cytokine production . Therefore , blocking TLR TIR dimerization may ameliorate O60603 -mediated hyperinflammatory states . The BB loop within the TLR TIR domain is critical for mediating certain protein-protein interactions . Examination of the human O60603 TIR domain crystal structure revealed a pocket adjacent to the highly conserved P681 and G682 BB loop residues . Using computer-aided drug design ( CADD ) , we sought to identify a small molecule inhibitor(s) that would fit within this pocket and potentially disrupt O60603 signaling . In silico screening identified 149 compounds and 20 US Food and Drug Administration-approved drugs based on their predicted ability to bind in the BB loop pocket . These compounds were screened in HEK293T- O60603 transfectants for the ability to inhibit O60603 -mediated P10145 mRNA . C16H15NO4 ( C29 ) was identified as a potential O60603 inhibitor . C29 , and its derivative , ortho-vanillin ( o-vanillin ) , inhibited O60603 /1 and O60603 /6 signaling induced by synthetic and bacterial O60603 agonists in human P29320 - O60603 and THP-1 cells , but only O60603 /1 signaling in murine macrophages . C29 failed to inhibit signaling induced by other TLR agonists and P01375 -α . Mutagenesis of BB loop pocket residues revealed an indispensable role for O60603 /1 , but not O60603 /6 , signaling , suggesting divergent roles . Mice treated with o-vanillin exhibited reduced O60603 -induced inflammation . Our data provide proof of principle that targeting the BB loop pocket is an effective approach for identification of O60603 signaling inhibitors . DB00731 and mitiglinide , but not sulfonylureas , induce insulin secretion through a mechanism mediated by calcium release from endoplasmic reticulum . DB00731 and mitiglinide ( glinides ) are characterized as rapid-onset and short-acting insulinotropic agents . Although both compounds do not have a sulfonylurea structure , it has been postulated that insulin secretion is preceded by their binding to Kir6.2/ Q09428 complex , and a mechanism of insulin secretion of glinides has been accounted for by this pathway . However , we hypothesized the involvement of additional mechanisms of insulin secretion enhanced by glinides , and we analyzed the pattern of time course of insulin secretion from MIN6 cells with the existence of agents that have specific pharmacologic actions . Dose-dependent effects of tolbutamide , glibenclamide , nateglinide , and mitiglinide were observed . P01308 secretion induced by 3 microM tolbutamide and 1 nM glibenclamide was completely inhibited by 10 microM diazoxide and 3 microM verapamil , although the latter half-component of insulin secretion profile induced by 3 microM nateglinide or 30 nM mitiglinide remained with the existence of those agents . Glinides enhanced insulin secretion even in Ca2+-depleted medium , and its pattern of secretion was same as the pattern with existence of verapamil . The latter half was suppressed by 1 microM dantrolene , and concomitant addition of verapamil and dantrolene completely suppressed the entire pattern of insulin secretion enhanced by nateglinide . Thus , we conclude that glinide action is demonstrated through two pathways , dependently and independently , from the pathway through K( DB00171 ) channels . We also demonstrated that the latter pathway involves the intracellular calcium release from endoplasmic reticulum via ryanodine receptor activation . Cellular distribution and contribution of cyclooxygenase P35354 to diabetogenesis in NOD mouse . Unlike most other mammalian cells , beta-cells of Langerhans constitutively express cyclooxygenase ( P36551 ) -2 rather than P23219 . P35354 is also constitutively expressed in type 1 diabetes ( T1D ) patients ' periphery blood monocytes and macrophage . To understand the role of P35354 in the beta-cell , we investigated P35354 expression in beta-cells and islet infiltrates of NOD and BALB/c mice using fluorescence immunohistochemistry and cytochemical confocal microscopy and Western blotting . Immunostaining showed that P35354 is expressed in islet-infiltrating macrophages , and that the expression of insulin and P35354 disappeared concomitantly from the beta-cells when NOD mice progressed toward overt diabetes . Also cultured P01308 -1E cells coexpressed insulin and P35354 but clearly in different subcellular compartments . Treatment with celecoxib increased insulin release from these cells in a dose-dependent manner in glucose concentrations ranging from 5 to 17 mM . Excessive P35354 expression by the islet-infiltrating macrophages may contribute to the beta-cell death during insulitis . The effects of celecoxib on P01308 -1E cells suggest that PGE(2) and other downstream products of P35354 may contribute to the regulation of insulin release from the beta-cells . Cost of community integrated prevention campaign for malaria , HIV , and diarrhea in rural Kenya . BACKGROUND : Delivery of community-based prevention services for HIV , malaria , and diarrhea is a major priority and challenge in rural Africa . Integrated delivery campaigns may offer a mechanism to achieve high coverage and efficiency . METHODS : We quantified the resources and costs to implement a large-scale integrated prevention campaign in Lurambi Division , Western Province , Kenya that reached 47,133 individuals ( and 83 % of eligible adults ) in 7 days . The campaign provided HIV testing , condoms , and prevention education materials ; a long-lasting insecticide-treated bed net ; and a water filter . Data were obtained primarily from logistical and expenditure data maintained by implementing partners . We estimated the projected cost of a Scaled-Up Replication ( Q09428 ) , assuming reliance on local managers , potential efficiencies of scale , and other adjustments . RESULTS : The cost per person served was $ 41.66 for the initial campaign and was projected at $ 31.98 for the Q09428 . The Q09428 cost included 67 % for commodities ( mainly water filters and bed nets ) and 20 % for personnel . The Q09428 projected unit cost per person served , by disease , was $ 6.27 for malaria ( nets and training ) , $ 15.80 for diarrhea ( filters and training ) , and $ 9.91 for HIV ( test kits , counseling , condoms , and P01730 testing at each site ) . CONCLUSIONS : A large-scale , rapidly implemented , integrated health campaign provided services to 80 % of a rural Kenyan population with relatively low cost . Scaling up this design may provide similar services to larger populations at lower cost per person . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Salmonid Tollip and MyD88 factors can functionally replace their mammalian orthologues in TLR-mediated trout P0DJI8 promoter activation . Many functional details of the piscine Toll-like receptor ( TLR ) signal-mediated activation of immune defense are still elusive . We used an established reconstitution system of mammalian TLR signaling to examine if this system would allow for pathogen-dependent promoter activation of the serum amyloid A ( P0DJI8 ) -encoding gene from rainbow trout ( Oncorhynchus mykiss ) and if the key mediators MyD88 and Tollip from trout can functionally substitute for their mammalian orthologues . Cells of the established human embryonic kidney line P29320 -293 were transiently co-transfected with vectors expressing bovine O60603 or O00206 factors and a reporter gene driven by the promoter of the trout P0DJI8 gene . Escherichia coli stimulation increased reporter gene expression more than 3-fold . Deletion series and point mutations identified in the proximal P0DJI8 promoter a composite overlapping binding site for NF-κB and P49715 factors as crucial for promoter activation . Overexpression of NF-κB p65 , but not of p50 or different members of the P49715 factor family proved this factor as an essential driver for P0DJI8 expression . Overexpression of a transdominant-negative mutant of the trout MyD88 factor reduced TLR-mediated P0DJI8 promoter activation confirming functional conservation of its TIR domain . Overexpression of the Tollip factor from trout also quenched TLR-mediated NF-κB and O00206 -mediated P0DJI8 promoter activation . The MyD88 mutant and Tollip expression studies confirm the functional homology of both piscine factors and their mammalian counterparts . We provide for the first time evidence that also the Tollip-mediated negative loop of TLR signaling may be conserved in non-mammalian organisms . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . Cyclophosphamide and other new agents for the treatment of severe aplastic anemia . Severe aplastic anemia ( P0DJI8 ) has a poor prognosis in the absence of treatment . Current accepted therapeutic strategies include allogeneic stem-cell transplantation and immunosuppression , both resulting in long-term survival in the majority of patients . Although human leukocyte antigen ( HLA ) -matched sibling stem-cell transplantation is highly effective , the 25 % probability of finding a suitable sibling donor within a family renders this approach available to only a minority of patients . Transplantation using HLA-matched , unrelated donors carries a high risk of treatment failure along with considerable toxicity . While combined immunosuppression with both antithymocyte globulin ( ATG ) and cyclosporine A ( Q13216 ) produces hematologic improvement in most patients , relapse is common . Late evolution of aplastic anemia to other serious hematologic disorders , including paroxysmal nocturnal hemoglobinuria ( PNH ) , myelodysplasia , and acute leukemia , is also a significant problem following treatment with ATG/ Q13216 . Recently , results of immunosuppression in P0DJI8 with another potent immunosuppressive agent , cyclophosphamide , were reported in a small number of patients . The overall response rate was similar to that seen with ATG/ Q13216 , but relapse and late clonal disease were not observed during a long period of follow-up . A larger randomized trial comparing sustained hematologic response rates to either conventional immunosuppression with ATG/ Q13216 or high-dose cyclophosphamide and Q13216 is now underway ; secondary end points include response duration , event-free survival , and overall survival . Additionally , a number of protocols designed to test the efficacy of alternative immunosuppressive or immunomodulatory agents are being developed . Development of stable cell lines expressing different subtypes of GABAA receptors . The experiments reported here were motivated by our interest to express in stably-transfected cells large amounts of recombinant rat GABAA receptors . For this , we developed an original two step selection strategy , in which the first step consisted of transfecting P29320 293 cells with rat GABAA receptor alpha and beta subunits . G 418 resistant colonies isolated at this step were screened for [ 3H ] muscimol binding to select for those that coexpressed alpha- and beta-subunits . The best alpha and beta subunit expressing colony was then supertransfected with a plasmid coding for the gamma rat GABAA receptor subunit and a mutant P00374 gene . After a second round of selection , this time in presence of methotrexate , those colonies that coexpressed ternary alpha beta gamma GABAA receptor combinations were distinguished using [ 3H ] flumazenil as a probe . This strategy was applied to the isolation of 3 GABAA receptor clones , alpha 1 beta 2 gamma 2s , alpha 3 beta 2 gamma 2s and alpha 5 beta 3 gamma 2s , that expressed relatively high levels of these proteins . These 3 cell lines exhibited pharmacological and functional properties similar to cells transiently-transfected with equivalent subunit combinations . These cell lines therefore provide attractive models with which to evaluate the intrinsic activity and potency of compounds at recombinant GABAA receptor subtypes . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Multiple single-nucleotide polymorphisms ( SNPs ) in the Japanese population in six candidate genes for long QT syndrome . We report here 20 single-nucleotide polymorphisms ( SNPs ) , including 15 novel ones , in six genes that are considered to be candidates for long QT syndrome ( LQTS ) : 2 SNPs in Q14721 , 3 in Q9UK17 , 3 in Q14654 , 7 in O60706 , 3 in P08588 , and 2 in Q05940 . We also examined their allelic frequencies in a Japanese sample population of LQTS-affected and nonaffected individuals . These data will be useful for genetic association studies designed to investigate acquired arrhythmias . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . Long-term follow-up after cyclophosphamide and cyclosporine-A therapy in steroid-dependent and -resistant nephrotic syndrome . A retrospective study was made on 37 children with idiopathic nephrotic syndrome ( P01308 ) . At the beginning , all patients were steroid sensitive but received more than one steroid course ( median 4 ) . Following several relapses , they became steroid dependent or steroid resistant . Group 1 consisted of 22 children [ 3 focal segmental glomerulosclerosis ( FSGS ) , 19 minimal-change NS ( MCNS ) ] who received cyclophosphamide ( CP ) orally for 2.5 +/- 0.5 months . Group 2 consisted of 15 children ( 7 FSGS , 8 MCNS ) who received cyclosporine-A ( Q13216 ) for 28 +/- 15 months . The level of proteinuria decreased significantly and remained low during the follow-up . The relapse-free period was significantly longer in the CP group ( CP 30 +/- 21.5 ; Q13216 26.2 +/- 18 months , p < 0.001 ) . The relapse rate decreased significantly in both groups and remained in this lower level during the follow-up ( from 3.4 +/- 2.8 to 0.1 +/- 0.2/year in group 1 , and from 3.7 +/- 3.1 to 0.6 +/- 0.8/year in group 2 ) . At the end of the 5-year follow-up , 20/22 patients ( 90.9 % ) and 10/15 patients ( 66.6 % ) were in remission in groups 1 and 2 respectively , with or without treatment ( p < 0.05 ) . In the long term , both CP and Q13216 is effective second-line therapy following steroid monotherapy in P01308 patients , but the relapse rate was lower and the relapse free period was significantly longer in the CP-treated group . Pharmacogenomic analysis of DB00171 -sensitive potassium channels coexpressing the common type 2 diabetes risk variants E23K and S1369A . OBJECTIVES : The common DB00171 -sensitive potassium ( KATP ) channel variants E23K and S1369A , found in the Q14654 and Q09428 genes , respectively , form a haplotype that is associated with an increased risk for type 2 diabetes . Our previous studies showed that KATP channel inhibition by the A-site sulfonylurea gliclazide was increased in the Q9C075 /A1369 haplotype . Therefore , we studied the pharmacogenomics of seven clinically used sulfonylureas and glinides to determine their structure-activity relationships in KATP channels containing either the E23/S1369 nonrisk or Q9C075 /A1369 risk haplotypes . RESEARCH DESIGN AND METHODS : The patch-clamp technique was used to determine sulfonylurea and glinide inhibition of recombinant human KATP channels containing either the E23/S1369 or the Q9C075 /A1369 haplotype . RESULTS : KATP channels containing the Q9C075 /A1369 risk haplotype were significantly less sensitive to inhibition by tolbutamide , chlorpropamide , and glimepiride ( IC50 values for Q9C075 /A1369 vs. E23/S1369=1.15 vs. 0.71 μmol/l ; 4.19 vs. 3.04 μmol/l ; 4.38 vs. 2.41 nmol/l , respectively ) . In contrast , KATP channels containing the Q9C075 /A1369 haplotype were significantly more sensitive to inhibition by mitiglinide ( IC50=9.73 vs. 28.19 nmol/l for Q9C075 /A1369 vs. E23/S1369 ) and gliclazide . DB00731 , glipizide , and glibenclamide showed similar inhibitory profiles in KATP channels containing either haplotype . CONCLUSION : Our results demonstrate that the ring-fused pyrrole moiety in several A-site drugs likely underlies the observed inhibitory potency of these drugs on KATP channels containing the Q9C075 /A1369 risk haplotype . It may therefore be possible to tailor existing therapy or design novel drugs that display an increased efficacy in type 2 diabetes patients homozygous for these common KATP channel haplotypes . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . P00797 -angiotensin system expression in rat bone marrow haematopoietic and stromal cells . The existence of a bone marrow renin-angiotensin system ( DB01367 ) is evidenced by the association of renin , angiotensin converting enzyme ( P12821 ) , and angiotensin ( Ang ) II and its AT(1) and AT(2) receptors with both normal and disturbed haematopoiesis . The expression of DB01367 components by rat unfractionated bone marrow cells ( BMC ) , haematopoietic-lineage BMC and cultured marrow stromal cells ( O60682 ) was investigated to determine which specific cell types may contribute to a local bone marrow DB01367 . The mRNAs for angiotensinogen , renin , P12821 , and AT(1a) and AT(2) receptors were present in BMC and in cultured O60682 ; Q9BYF1 mRNA was detected only in BMC . Two-colour flow fluorocytometry analysis showed immunodetectable angiotensinogen , P12821 , AT(1) and AT(2) receptors , and Ang II , as well as binding of Ang II to AT(1) and AT(2) receptors , in P01730 (+) , CD11b/c(+) , CD45R(+) and CD90(+) BMC and cultured O60682 ; renin was found in all cell types with the exception of P01730 (+) BMC . Furthermore , Ang II was detected by radioimmunoassay in O60682 homogenates as well as conditioned culture medium . The presence of Ang II receptors in both haematopoietic-lineage BMC and O60682 , and the de novo synthesis of Ang II by O60682 suggest a potential autocrine-paracrine mechanism for local DB01367 -mediated regulation of haematopoiesis . Permanent neonatal diabetes mellitus in China . BACKGROUND : Permanent neonatal diabetes mellitus ( PNDM ) is a rare disease , which is defined as the onset of diabetes before the age of 6 months with persistence through life . Infants with Q14654 or Q09428 genetic mutations may respond to oral sulfonylurea therapy . Currently , there are limited studies about the genetic analysis and long-term follow-up of PNDM . CASE PRESENTATION : We report four cases of PNDM . None of the infants or their parents had P01308 , Q14654 , or Q09428 genetic mutations . One infant underwent continuous subcutaneous insulin infusion ( CSII ) and the other infants underwent multiple injections of insulin ( MII ) . In these infants , PNDM persisted from 35 months to 60 months of follow-up . Three infants maintained fairly stable blood sugar levels , and one infant had poor sugar control . CONCLUSIONS : We suggest that all of the infants with PNDM should undergo genetic evaluation . For infants without Q14654 and Q09428 genetic mutations , oral sulfonylurea should not be considered as treatment . CSII is a useful method for overcoming the difficulties of diabetes , and it may also improve the quality of life of both infants and their parents . P01308 -like growth factor-1 receptor inhibitor , Q99217 -479 , in cetuximab-refractory head and neck squamous cell carcinoma . BACKGROUND : Recurrent head and neck squamous cell carcinoma ( HNSCC ) remains a difficult cancer to treat . Here , we describe a patient with HNSCC who had complete response to methotrexate ( MTX ) after progressing on multiple cytotoxic agents , cetuximab , and Q99217 -479 ( monoclonal antibody against insulin-like growth factor-1 receptor [ IGF-1R ] ) . METHODS : The clinical information was collected by a retrospective medical record review under an Institutional Review Board-approved protocol . From 4 tumors and 2 normal mucosal epithelia , global gene expression , and IGF-1R and dihydrofolate reductase ( P00374 ) protein levels were determined . RESULTS : Effective target inhibition in the tumor was confirmed by the decreased protein levels of total and phospho-IGF-1R after treatment with Q99217 -479 . Decreased level of P00374 and conversion of a gene expression profile associated with cetuximab-resistance to cetuximab-sensitivity were also observed . CONCLUSION : This suggests that the combination of Q99217 -479 and MTX or cetuximab may be a promising therapeutic approach in refractory HNSCC . Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 -induced survival . This inhibition can not be overcome by increasing concentrations of P05113 and is not due to the blocking of Na+ channels by lidocaine . Here we report that one class of K+ channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 . In contrast , increasing concentrations of P08700 or granulocyte-macrophage P04141 overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 -sensitive K+ channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [3H]glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . Differential interactions of nateglinide and repaglinide on the human beta-cell sulphonylurea receptor 1 . DB00912 and nateglinide represent a new class of insulin secretagogues , structurally unrelated to sulphonylureas , that were developed for the treatment of type 2 diabetes . The inhibitory effect of these drugs was investigated on recombinant wild-type and mutant Kir6.2/ Q09428 channels expressed in HEK293 cells . DB00731 and repaglinide dose-dependently inhibited whole-cell Kir6.2/ Q09428 currents with half-maximal inhibitory concentration ( IC(50) ) values of 800 and 21 nmol/l , respectively . Mutation of serine 1237 in Q09428 to tyrosine ( S1237Y ) abolished tolbutamide and nateglinide block , suggesting that these drugs share a common point of interaction on the Q09428 subunit of the DB00171 -sensitive K(+) channel . In contrast , repaglinide inhibition was unaffected by the S1237Y mutation ( IC(50) = 23 nmol/l ) . Radioligand binding studies revealed a single high-affinity binding site for [(3)H]repaglinide on membranes prepared from HEK293 cells expressing wild-type ( equilibrium dissociation constant [ K(D) ] = 0.40 nmol/l ) or mutant ( K(D) = 0.31 nmol/l ) Kir6.2/ Q09428 channels . DB00731 and tolbutamide displaced [(3)H]repaglinide binding to wild-type channels with IC(50) values of 0.7 and 26 micro mol/l , respectively , but produced < 10 % displacement of [(3)H]repaglinide bound to mutant channels . This is consistent with the idea that binding of nateglinide and tolbutamide , but not repaglinide , is abolished by the Q09428 [S1237Y] mutation and that the binding site for repaglinide is not identical to that of nateglinde/tolbutamide . These results are discussed in terms of a conformational analysis of the drug molecules . P01308 resistance and angiotensin converting enzyme polymorphism in Japanese hypertensive subjects . P00797 -angiotensin system activity has been shown to affect insulin sensitivity . However , the relationship between I/D polymorphism and insulin resistance is controversial . Therefore , we examined the relationship between the P12821 genotype and insulin sensitivity in 51 Japanese hypertensive patients using the glucose clamp technique . The P12821 genotype distribution in the hypertensive subjects was : 7 subjects with DD , 20 subjects with ID , and 24 subjects with II . P01308 sensitivity in terms of the glucose disposal rate was not significantly different among the three P12821 genotypes , although there was a tendency for insulin sensitivity to decrease in the order of II , ID and DD , DD being the lowest . These findings are contrary to previous reports that insulin sensitivity was increased in normotensive subjects with the DD genotype who were Caucasian or African-American . There might be a difference due to race and whether the subjects are hypertensive or obese . We concluded that insulin sensitivity was not different among the P12821 genotypes in the Japanese hypertensive subjects , supporting a previous report on the Chinese population . To date , insulin sensitivity has not been found to differ with P12821 genotypes in the oriental population . Two-dimensional liquid crystalline growth within a phase-field-crystal model . By using a two-dimensional phase-field-crystal ( P27918 ) model , the liquid crystalline growth of the plastic triangular phase is simulated with emphasis on crystal shape and topological defect formation . The equilibrium shape of a plastic triangular crystal ( PTC ) grown from an isotropic phase is compared with that grown from a columnar or smectic-A ( Q13216 ) phase . While the shape of a PTC nucleus in the isotropic phase is almost identical to that of the classical P27918 model , the shape of a PTC nucleus in Q13216 is affected by the orientation of stripes in the Q13216 phase , and irregular hexagonal , elliptical , octagonal , and rectangular shapes are obtained . Concerning the dynamics of the growth process , we analyze the topological structure of the nematic order , which starts from nucleation of +1/2 and -1/2 disclination pairs at the PTC growth front and evolves into hexagonal cells consisting of +1 vortices surrounded by six satellite -1/2 disclinations . It is found that the orientational and the positional order do not evolve simultaneously ; the orientational order evolves behind the positional order , leading to a large transition zone , which can span over several lattice spacings . DB09026 : An orally active renin inhibitor . P00797 inhibitors are antihypertensive drugs that block the first step in the renin-angiotensin system . Their mechanism of action differs from that of the angiotensin-converting enzyme inhibitors and angiotensin-receptor antagonists , but like these drugs , renin inhibitors interrupt the negative feedback effects of angiotensin II on renin secretion . The renin-angiotensin-aldosterone system ( RAAS ) has long been recognized to play a significant role in hypertension pathophysiology . Certain agents that modify the RAAS can control blood pressure and improve cardiovascular outcomes . Optimization of this compound by Novartis led to the development of aliskiren - the only direct renin inhibitor which is clinically used as an antihypertensive drug . DB09026 is the first of a new class of antihypertensive agents . DB09026 is a new renin inhibitor of a novel structural class that has recently been shown to be efficacious in hypertensive patients after once-daily oral dosing . In short-term studies , it was effective in lowering blood pressure either alone or in combination with valsartan and hydrochlorothiazide , and had a low incidence of serious adverse effects . It was approved by the Food and Drug Administration in 2007 for the use as a monotherapy or in combination with other antihypertensives . Greater reductions in blood pressure have been achieved when aliskiren was used in combination with hydrochlorothiazide or an angiotensin-receptor blocker . The most common adverse effects reported in clinical trials were headache , fatigue , dizziness , diarrhea , and nasopharyngitis . DB09026 has not been studied in patients with moderate renal dysfunction ; as an RAAS-acting drug , it should be prescribed for such patients only with caution . HRAS1 and P01308 genes are relocated but not structurally altered as a result of the t(7;11)(p15;p15) in a clone from a patient with acute myeloid leukaemia ( M4 ) . A patient whose leukaemic cells carried the rare t(7;11)(p15;p15) was diagnosed as having acute myelomonocytic leukaemia ( AML-M4 ) , and supports the association of this specific translocation with forms of acute myeloid leukaemia showing differentiation . Blast phase chronic myeloid leukaemia was excluded by lack of involvement of the P00519 and P11274 genes . Chromosome in situ hybridization studies showed that both the HRAS1 and P01308 genes were present on the terminal part of chromosome 11p which was translocated to chromosome 7p . Neither HRAS1 nor P01308 were structurally rearranged . Field inversion gel electrophoresis showed that a 400 kb fragment encompassing HRAS1 was structurally entire in leukaemic DNA . Because the P01308 gene , which was also translocated , is probably located proximal to HRAS1 on chromosome 11p , it is unlikely that HRAS1 was near the chromosome 11 breakpoint or involved in this leukaemia . Recent therapeutic strategy for sustained ventricular tachycardia in Japan . We investigated the therapeutic principles and strategies to treat sustained ventricular tachycardia ( SVT ) as five leading medical institutions in the Tokyo area and summarized the present situation of SVT treatment in Japan . Catheter ablation ( P00519 ) has been almost established to be effective in idiopathic ventricular tachycardia ( IVT ) and was used as the last treatment in 60.3 % of IVTs in this series . P00519 may be the first option of therapy for IVT . In patients with SVT who have underlying cardiac diseases , the last treatment was class I drugs in 8.3 % , class III drugs in 34.3 % , combination drug therapy in 24.0 % , P00519 in 33.3 % , surgical therapy ( Q09428 ) in 7.3 % , and implantable cardioverter defibrillator ( ICD ) in 12.5 % ( nonpharmacological therapy in combination with other therapy ) . The use of class I drugs was not common , whereas class III drugs were used more frequently in patients with a low left ventricular ejection fraction . In some patients with reduced cardiac function , a combination of class III drugs and non-pharmacological therapy is appropriate . Physical and functional interaction between the dopamine transporter and the synaptic vesicle protein synaptogyrin-3 . Uptake through the dopamine transporter ( Q01959 ) represents the primary mechanism used to terminate dopaminergic transmission in brain . Although it is well known that dopamine ( DA ) taken up by the transporter is used to replenish synaptic vesicle stores for subsequent release , the molecular details of this mechanism are not completely understood . Here , we identified the synaptic vesicle protein synaptogyrin-3 as a Q01959 interacting protein using the split ubiquitin system . This interaction was confirmed through coimmunoprecipitation experiments using heterologous cell lines and mouse brain . Q01959 and synaptogyrin-3 colocalized at presynaptic terminals from mouse striatum . Using fluorescence resonance energy transfer microscopy , we show that both proteins interact in live neurons . Pull-down assays with Q86UG4 ( glutathione S-transferase ) proteins revealed that the cytoplasmic N termini of both Q01959 and synaptogyrin-3 are sufficient for this interaction . Furthermore , the N terminus of Q01959 is capable of binding purified synaptic vesicles from brain tissue . Functional assays revealed that synaptogyrin-3 expression correlated with Q01959 activity in PC12 and MN9D cells , but not in the non-neuronal P29320 -293 cells . These changes were not attributed to changes in transporter cell surface levels or to direct effect of the protein-protein interaction . Instead , the synaptogyrin-3 effect on Q01959 activity was abolished in the presence of the vesicular monoamine transporter-2 ( Q05940 ) inhibitor reserpine , suggesting a dependence on the vesicular DA storage system . Finally , we provide evidence for a biochemical complex involving Q01959 , synaptogyrin-3 , and Q05940 . Collectively , our data identify a novel interaction between Q01959 and synaptogyrin-3 and suggest a physical and functional link between Q01959 and the vesicular DA system . Effect of acetazolamide on aquaporin-1 and fluid flow in cultured choroid plexus . DB00819 ( AZA ) , used in treatment of early or infantile hydrocephalus , is effective in some cases , while its effect on the choroid plexus ( CP ) remains ill-defined . The drug reversibly inhibits aquaporin-4 ( P55087 ) , the most ubiquitous " water pore " in the brain , and perhaps modulation of P29972 ( located apically on CP cells ) by AZA may reduce cerebrospinal fluid ( P04141 ) production . We sought to elucidate the effect of AZA on P29972 and fluid flow in CP cell cultures.CP tissue culture from 10-day Sprague-Dawley rats and a TRCSF-B cell line were grown on Transwell permeable supports and treated with 100 μM AZA . Fluid assays to assess direction and extent of fluid flow , and P29972 expression patterns by immunoblot , Immuncytochemistry ( ICC ) , and quantitative reverse transcriptase polymerase chain reaction ( qRT-PCR ) were performed.Immunoblots and ICC analyses showed a decrease in P29972 protein shortly after AZA treatment ( lowest at 12 h ) , with transient P29972 reduction mediated by mRNA expression ( lowest at 6 h ) . Transwell fluid assays indicated a fluid shift at 2 h , before significant changes in P29972 mRNA or protein levels.Timing of AZA effect on P29972 suggests the drug alters protein transcription , while affecting fluid flow by a concomitant method . It is plausible that other mechanisms account for these phenomena , as the processes may occur independently . Domain organization of the DB00171 -sensitive potassium channel complex examined by fluorescence resonance energy transfer . K( DB00171 ) channels link cell metabolism to excitability in many cells . They are formed as tetramers of Kir6.2 subunits , each associated with a Q09428 subunit . We used mutant GFP-based FRET to assess domain organization in channel complexes . Q8N1N2 -length Kir6.2 subunits were linked to YFP or cyan fluorescent protein ( P27918 ) at N or C termini , and all such constructs , including double-tagged YFP-Kir6.2- P27918 ( Y6.2C ) , formed functional K( DB00171 ) channels . In intact COSm6 cells , background emission of YFP excited by 430-nm light was ∼6 % , but the Y6.2C construct expressed alone exhibited an apparent FRET efficiency of ∼25 % , confirmed by trypsin digestion , with or without Q09428 co-expression . Similar FRET efficiency was detected in mixtures of P27918 - and YFP-tagged full-length Kir6.2 subunits and transmembrane domain only constructs , when tagged at the C termini but not at the N termini . The FRET-reported Kir6.2 tetramer domain organization was qualitatively consistent with Kir channel crystal structures : C termini and M2 domains are centrally located relative to N termini and M1 domains , respectively . Additional FRET analyses were performed on cells in which tagged full-length Kir6.2 and tagged Q09428 constructs were co-expressed . These analyses further revealed that 1 ) NBD1 of Q09428 is closer to the C terminus of Kir6.2 than to the N terminus ; 2 ) the Kir6.2 cytoplasmic domain is not essential for complexation with Q09428 ; and 3 ) the N-terminal half of Q09428 can complex with itself in the absence of either the C-terminal half or Kir6.2 . Chemically synthesized SDF-1alpha analogue , N33A , is a potent chemotactic agent for P61073 / P61073 / P61073 -expressing human leukocytes . Stromal cell-derived factor ( SDF ) 1 is a potent chemoattractant for leukocytes through activation of the receptor P61073 / P61073 / P61073 , which is a fusion co-factor for the entry of T lymphocytotropic human immunodeficiency virus type 1 ( HIV-1 ) . This P61073 -mediated HIV-1 fusion can be inhibited by P48061 . Because of its importance in the study of immunity and AIDS , large scale production of P48061 is desirable . In addition to recombinant technology , chemical synthesis provides means by which biologically active proteins can be produced not only in large quantity but also with a variety of designed modifications . In this study , we investigated the binding and function of an SDF-1alpha analogue , N33A , synthesized by a newly developed native chemical ligation approach . Radioiodinated N33A showed high affinity binding to human monocytes , T lymphocytes , as well as neutrophils , and competed equally well with native recombinant SDF-1alpha for binding sites on leukocytes . N33A also showed equally potent chemoattractant activity as native recombinant SDF-1alpha for human leukocytes . Further study with P61073 / P61073 / P61073 transfected P29320 293 cells showed that N33A binds and induces directional migration of these cells in vitro . These results demonstrate that the chemically synthesized SDF-1alpha analogue , N33A , which can be produced rapidly in large quantity , possesses the same capacity as native SDF-1alpha to activate P61073 -expressing cells and will provide a valuable agent for research on the host immune response and AIDS . The effects of mitiglinide ( KAD-1229 ) , a new anti-diabetic drug , on DB00171 -sensitive K+ channels and insulin secretion : comparison with the sulfonylureas and nateglinide . DB01252 ( KAD-1229 ) , a new anti-diabetic drug , is thought to stimulate insulin secretion by closing the DB00171 -sensitive K+ ( K( DB00171 ) ) channels in pancreatic beta-cells . However , its selectivity for the various K( DB00171 ) channels is not known . In this study , we examined the effects of mitiglinide on various cloned K( DB00171 ) channels ( Kir6.2/ Q09428 , Kir6.2/SUR2A , and Kir6.2/SUR2B ) reconstituted in COS-1 cells , and compared them to another meglitinide-related compound , nateglinide . Patch-clamp analysis using inside-out recording configuration showed that mitiglinide inhibits the Kir6.2/ Q09428 channel currents in a dose-dependent manner ( IC50 value , 100 nM ) but does not significantly inhibit either Kir6.2/SUR2A or Kir6.2/SUR2B channel currents even at high doses ( more than 10 microM ) . DB00731 inhibits Kir6.2/ Q09428 and Kir6.2/SUR2B channels at 100 nM , and inhibits Kir6.2/SUR2A channels at high concentrations ( 1 microM ) . Binding experiments on mitiglinide , nateglinide , and repaglinide to Q09428 expressed in COS-1 cells revealed that they inhibit the binding of [3H]glibenclamide to Q09428 ( IC50 values : mitiglinide , 280 nM ; nateglinide , 8 microM ; repaglinide , 1.6 microM ) , suggesting that they all share a glibenclamide binding site . The insulin responses to glucose , mitiglinide , tolbutamide , and glibenclamide in MIN6 cells after chronic mitiglinide , nateglinide , or repaglinide treatment were comparable to those after chronic tolbutamide and glibenclamide treatment . These results indicate that , similar to the sulfonylureas , mitiglinide is highly specific to the Kir6.2/ Q09428 complex , i.e. , the pancreatic beta-cell K( DB00171 ) channel , and suggest that mitiglinide may be a clinically useful anti-diabetic drug . Single nucleotide polymorphisms associated with aggressive periodontitis and severe chronic periodontitis in Japanese . Periodontitis is a common inflammatory disease causing destruction of periodontal tissues . It is a multifactor disease involving genetic factors and oral environmental factors . To determine genetic risk factors associated with aggressive periodontitis or severe chronic periodontitis , single nucleotide polymorphisms ( SNPs ) in multiple candidate genes were investigated in Japanese . We studied 134 patients with aggressive periodontitis , 117 patients with severe chronic periodontitis , and 125 healthy volunteers without periodontitis , under case-control setting , and 310 SNPs in 125 candidate genes were genotyped . Association evaluation by Fisher 's exact test ( p < 0.01 ) revealed statistically significant SNPs in multiple genes , not only in inflammatory mediators ( P40189 and P41222 , associated with aggressive periodontitis ; and P07339 , associated with severe chronic periodontitis ) , but also in structural factors of periodontal tissues ( P02462 , P02452 , and Q9C075 , associated with aggressive periodontitis ; and P98160 , Q9UMD9 , and P01133 , associated with severe chronic periodontitis ) . These appear to be good candidates as genetic factors for future study . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . DB00731 is Effective for Diabetes Mellitus with Reactive Hypoglycemia in a Child with a Compound Heterozygous Q09428 Mutation . Q09428 encodes the sulfonylurea receptor 1 ( Q09428 ) subunits of the beta-cell DB00171 -sensitive potassium ( K- DB00171 ) channel playing a critical role in the regulation of insulin secretion , and inactivating mutations in Q09428 cause congenital hyperinsulinism . Recently , Q09428 inactivating mutations were reported to be involved in the development of diabetes mellitus later in life . We report a girl who was born macrosomic with transient hypoglycemia and thereafter developed diabetes mellitus accompanied by severe reactive hypoglycemia at the age of 11 yr . An OGTT ( oral glucose tolerance test ) revealed hyperglycemia due to poor early insulin response and subsequent hypoglycemia due to delayed prolonged insulin secretion . Hypoglycemia was improved by the combination of nateglinide , which stimulates early insulin secretion , and an alpha-glucosidase inhibitor , voglibose . Sequencing of the Q09428 identified a compound heterozygous mutation ( R1420H/F591fs604X ) , suggesting that this mutation may alter regulation of insulin secretion with advancing age , leading to diabetes mellitus with reactive hypoglycemia from hyperinsulinism . Therefore , long-term follow-up and periodic OGTTs are important for early detection of insulin dysregulation in congenital hyperinsulinism patients carrying the Q09428 mutation , even though hypoglycemia resolves spontaneously during infancy . Furthermore , nateglinide may be useful therapeutically in the treatment of not only diabetes mellitus but also reactive hypoglycemia . DB06756 modulates age-related NF-kappaB by thiol-enhancing action . Depletion of glutathione levels and perturbations in redox status are considered to play a crucial role in aging and chronic inflammatory processes through the activation of redox sensitive transcription factors , including nuclear factor-kappaB ( NF-kappaB ) . In the current study , we assessed the regulatory action of dietary betaine in the suppression of NF-kappaB by comparing kidney tissue from old , betaine-supplemented rats or non-betaine-supplemented rats ( age 21 months ) and 7 month-old rats . In addition , cultured P29320 293T cells were utilized for the molecular assessment of betaine 's restorative ability of redox status when treating cells with potent glutathione ( DB00143 ) -depleting agents . Results showed that in old rats a short-term feeding ( 10 d ) with betaine attenuated the age-related decrease in thiol levels , increase in reactive species and TNFalpha expression via NF-kappaB activation , compared to the young controls . These findings were verified in the cell-cultured system . Further investigations found that redox imbalance due to thiol depletion caused increased NF-kappaB activation , and cyclooxygenase ( P36551 ) -2 and TNFalpha levels , both of which were suppressed by betaine treatment . Based on both in vivo and in vitro data , we concluded that betaine exerts its efficacy by maintaining thiol status in the regulation of P35354 and TNFalpha via NF-kappaB activation during aging . Methylation of promoter regions of genes of the human intrauterine P00797 Angiotensin system and their expression . The intrauterine renin angiotensin system ( DB01367 ) is implicated in placentation and labour onset . Here we investigate whether promoter methylation of DB01367 genes changes with gestation or labour and if it affects gene expression . Early gestation amnion and placenta were studied , as were term amnion , decidua , and placenta collected before labour ( at elective caesarean section ) or after spontaneous labour and delivery . The expression and degree of methylation of the prorenin receptor ( O75787 ) , angiotensin converting enzyme ( P12821 ) , angiotensin II type 1 receptor ( P30556 ) , and two proteases that can activate prorenin ( kallikrein , P06870 , and cathepsin D , P07339 ) were measured by qPCR and a DNA methylation array . There was no effect of gestation or labour on the methylation of DB01367 genes and P07339 . Amnion and decidua displayed strong correlations between the percent hypermethylation of DB01367 genes and P07339 , suggestive of global methylation . There were no correlations between the degree of methylation and mRNA abundance of any genes studied . P06870 was the most methylated gene and the proportion of hypermethylated P06870 alleles was lower in placenta than decidua . The presence of intermediate methylated alleles of P06870 in early gestation placenta and in amnion after labour suggests that P06870 methylation is uniquely dynamic in these tissues . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells . We evaluated the effect of islet neogenesis-associated protein pentadecapeptide ( INGAP-PP ) upon islet beta- and non-beta cell differentiation from mouse embryonic stem ( mES ) cells . ES-D3 cell lines were cultured following Lumelsky 's protocol with or without INGAP-PP ( 5 microg/ml ) at different stages . Gene expression was quantified using qPCR. mES cells were fixed and immunostained using anti insulin- , somatostatin- , glucagon- , Pdx-1- , Ngn-3- , Nkx-6.1 and P09936 specific antibodies . P12004 was used to measure replication rate . Bcl(2) ( immunostaining ) and caspase-3 ( enzyme activity and gene expression ) were determined as apoptosis markers . INGAP-PP increased P10997 , Glut-2 , Kir-6.2 , Q09428 -1 and insulin gene expression , and the percentage of insulin-immunostained cells . Conversely , INGAP-PP reduced significantly glucagon and somatostatin gene expression and immunopositivity . While nestin gene expression was not affected , there was a significant reduction in the percentage of P09936 -immunostained cells . Pdx-1 gene expression increased by 115 % in INGAP-PP treated cells , as well as the percentage of Pdx-1 , Ngn-3 and Nkx-6.1 immunopositive cells . Neither caspase-3 ( expression and activity ) nor Bcl(2) positively immunostained cells were affected by INGAP-PP . Accordingly , INGAP-PP would promote stem cell differentiation into a beta-like cell phenotype , simultaneously decreasing its differentiation toward non-beta-cell precursors . Therefore , INGAP-PP would be potentially useful to obtain beta-cells from stem cells for replacement therapy . [ Changes in chemokine receptor 4 , interleukin-6 , and collagen X expression in the ATDC5 cell line stimulated by cyclic tensile strain and stromal cell-derived factor-1 ] . OBJECTIVE : This study further explores the stromal cell-derived factor-1 ( P48061 ) /chemokine receptor 4 ( P61073 ) signaling axis mechanism in temporomandibular joint osteoarthritis ( OA ) by detecting the changes in P61073 , interleukin ( IL ) -6 , and collagen X expression in the ATDC5 cell line stimulated by the cyclic tensile strain and P48061 . METHODS : P01308 -transferrin-selenium ( ITS ) was used to induce ATDC5 cells to differentiate into chondrocyte-like cells . After three weeks , the cells were divided into two groups : those with and without cyclic tensile strain . These groups were further divided into the negative control and P48061 groups . Strain force of 20 % was applied . After 12 h , the total proteins were extracted from cells of the four groups , and Western blot analysis was used to detect the changes in P61073 , P05231 , and collagen X expression . RESULTS : P48061 could enhance P61073 , P05231 , and collagen X expressions in the chondrocytes , and 20 % tensile strain force could further upregulate the three factors . CONCLUSION : Under abnormal tensile force , P48061 can upregulate its specific receptor P61073 , thus increasing its-binding efficiency and resulting in the activation of the P48061 / P61073 axis . This condition enhances the expressions of P05231 and other inflammatory factors and directly damages to cartilage tissue . Such damage directly promotes chondrocyte hypertrophy , which enhances collagen X expression . Effect of long-term type 1 diabetes on renal sodium and water transporters in rats . The effects of long-term diabetes in the presence of established nephropathy on tubular function remains poorly understood . We evaluated the levels of the main sodium and water transport proteins expressed in the kidney after long-term ( 8 weeks ) of streptozotocin ( Q11206 ) -induced type 1 diabetes mellitus ( DM ) in untreated ( D ) and insulin ( 4 U/s.c./day ) -treated ( D+I ) rats . D animals presented upregulation ( approximately 4.5-fold ) of Na/glucose cotransporter ( P13866 ) , whereas the alpha-subunit of the epithelial sodium channel ( alpha-ENaC ) and aquaporin 1 ( P29972 ) were downregulated ( approximately 20 and 30 % respectively ) with no change in the Na/H exchanger ( P48764 ) , Na/Cl cotransporter ( TSC ) and P41181 . P01308 replacement partially prevented these alterations and caused increases in the expression of alpha-ENaC and P41181 . These effects suggest an action of insulin in the tubular transport properties . The upregulation of P13866 may constitute a mechanism to prevent greater glucose losses in the urine but it may result in glucotoxicity to the proximal epithelial cells contributing to the diabetic nephropathy . The decrease of alpha-ENaC in D animals may compensate for the increased sodium reabsorption via P13866 resulting in discrete natriuresis . DM-induced polyuria was not due to changes in P41181 expression . Pancreatic beta-cell K( DB00171 ) channel activity and membrane-binding studies with nateglinide : A comparison with sulfonylureas and repaglinide . DB00731 ( A-4166 ) is an amino acid derivative with insulinotrophic action in clinical development for treatment of type 2 diabetes . The aim of this study was to determine whether nateglinide 's interaction at the K( DB00171 ) channel/sulfonylurea receptor underlies its more rapid onset and shorter duration of action in animal models . Binding studies were carried out with membranes prepared from Q99578 -m5F cells and P29320 -293 cells expressing recombinant human sulfonylurea receptor 1 ( Q09428 ) . The relative order for displacement of [(3)H]glibenclamide in competitive binding experiments with Q99578 -m5F cell membranes was glibenclamide > glimepiride > repaglinide > glipizide > nateglinide > L-nateglinide > tolbutamide . The results with P29320 -293/recombinant human Q09428 cells were similar with the exception that glipizide was more potent than repaglinide . Neither nateglinide nor repaglinide had any effect on the dissociation kinetics for [(3)H]glibenclamide , consistent with both compounds competitively binding to the glibenclamide-binding site on Q09428 . Finally , the inability to measure [(3)H]nateglinide binding suggests that nateglinide dissociates rapidly from Q09428 . Direct interaction of nateglinide with K( DB00171 ) channels in rat pancreatic beta-cells was investigated with the patch-clamp method . The relative potency for inhibition of the K( DB00171 ) channel was repaglinide > glibenclamide > nateglinide . Kinetics of the inhibitory effect on K( DB00171 ) current showed that the onset of inhibition by nateglinide was comparable to glibenclamide but more rapid than that of repaglinide . The time for reversal of channel inhibition by nateglinide was also faster than with glibenclamide and repaglinide . These results suggest that the unique characteristics of nateglinide are largely the result of its interaction at the K( DB00171 ) channel . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity .

MH_train_27

MH_train_27

MH_train_27

interacts_with DB09045?

multiple_choice

Adulthood nicotine treatment alleviates behavioural impairments in rats neonatally treated with quinpirole : possible roles of acetylcholine function and neurotrophic factor expression . Increases in dopamine D(2) receptor sensitivity are known to be common in drug abuse and neurological disorders . Past data from this laboratory have shown that long-term increases in D(2) sensitivity can be produced by quinpirole treatment ( a D(2)/D(3) agonist ) during early development . The present investigation was designed to test the hypothesis that nicotine administration in adulthood would reduce both cognitive and skilled reaching impairments produced by increases in D(2) sensitivity . Female Sprague-Dawley rats were treated with quinpirole ( 1 mg/kg ) or saline from postnatal day 1 ( PD 1 ) to PD 21 . Beginning in adulthood ( PD 61 ) , rats were treated with nicotine ( 0.3 mg/kg free base ) or saline twice daily for 14 consecutive days before behavioural testing commenced . Animals neonatally treated with quinpirole demonstrated performance deficits on the Morris water task and a skilled reaching task compared to controls . Deficits on both tasks were completely alleviated by adulthood nicotine treatment . Animals neonatally treated with quinpirole demonstrated a significant 36 % decrease of P28329 in the hippocampus compared to saline controls that was partially eliminated by nicotine . Additionally , neonatal quinpirole produced a significant decrease in hippocampal P01138 content compared to controls , however , nicotine failed to alleviate this decrease in P01138 . The results of this investigation demonstrate that long-term increases in dopamine D(2) receptor sensitivity produce significant decreases in hippocampal cholinergic and P01138 expression that may result in cognitive impairment . DB00184 alleviates both cognitive and skilled reaching impairments caused by increases in D(2) sensitivity , but the mechanism through which nicotine is acting is currently unknown . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . P01258 stimulates the secretion of urokinase-type plasminogen activator from prostate cancer cells : its possible implications on tumor cell invasion . P01258 ( CT ) is synthesized and secreted in prostate epithelium , and its secretion from malignant prostates is several folds higher than that in benign prostates . CT receptor ( CTR ) is expressed in malignant prostate epithelium , and its activation increases invasiveness of prostate cancer ( PC ) cells via activation of protein kinase A . Since the role of urokinase-type plasminogen activator ( uPA ) in invasion of PC has been established , we tested the hypothesis that CT increases invasion of PC cells by stimulating uPA secretion from PC cells . Exogenously added CT stimulated the secretion of uPA from PC-3M cells in a dose-dependent manner , which was blocked by Rp. DB02527 , a competitive inhibitor of protein kinase A . CT stimulated the secretion of P08253 and P14780 from PC-3M cells , and also increased their invasiveness . Both these actions of CT were blocked by uPA-neutralizing antibodies . Immunofluorescence studies with PC-3M cells suggest that CT stimulated redistribution of cellular uPA to focal adhesion sites , which was further confirmed by co-immunoprecipitation of uPA with focal adhesion kinase ( Q05397 ) in response to CT . These results suggest that CT increases invasiveness of PC cells by stimulating PKA-mediated uPA secretion and by redirecting the secreted uPA to focal adhesion sites . The results also suggest that uPA may , at least in part , mediate proinvasive actions of CT on PC cells by stimulating the secretion of gelatinases and degradation of focal adhesion sites . Glucagon-like peptide-1(1-37) inhibits chemokine-induced migration of human P01730 -positive lymphocytes . The present study examined the effect of P0C6A0 (1-37) on chemokine-induced P01730 -positive lymphocyte migration as an early and critical step in atherogenesis . Pretreatment with P0C6A0 (1-37) reduced the SDF-induced migration of isolated human P01730 -positive lymphocytes in a concentration-dependent manner . Similar effects were seen when RANTES was used as a chemokine . P0C6A0 (1-37) 's effect on P01730 -positive lymphocyte migration was mediated through an early inhibition of chemokine-induced P19957 kinase activity . Downstream , P0C6A0 (1-37) inhibited SDF-induced phosphorylation of MLC and cofilin and limited f-actin formation as well as P32942 translocation . Furthermore , exendin-4 inhibited SDF-induced migration of P01730 -positive lymphocytes similarly to P0C6A0 (1-37) , and transfection of these cells with P43220 siRNA abolished P0C6A0 (1-37) 's action on chemokine-induced P32942 translocation , suggesting an effect mediated via the P43220 . Thus , P0C6A0 (1-37) inhibits chemokine-induced P01730 -positive lymphocyte migration by inhibition of the P19957 -kinase pathway and via the P43220 . This effect provides a potential novel mechanism for how P0C6A0 (1-37) may modulate vascular disease . Exendin-4 exerts its effects through the P01138 /p75NTR system in diabetic mouse pancreas . Glucagon-like peptide-1 ( P0C6A0 ) ameliorates the symptoms of diabetes through stimulation of insulin secretion . We have investigated the possible components of cellular mechanism triggered by exendin-4 , a potent P43220 agonist , in streptozotocin ( Q11206 ) induced diabetic mice pancreas . BALB/c male mice were divided into four groups for this investigation . The first group was given citrate buffer only , the second group was administered exendin-4 alone , the third group received Q11206 , and the fourth group was given both Q11206 and exendin-4 . Exendin-4 ( 3 microg/kg ) was administered by daily subcutaneous injection for 30 days after the animals were rendered diabetic by administration of Q11206 ( 200 mg/kg ) . With exendin-4 treatment on diabetic mice , the following results were noted : ( i ) exendin-4 suppressed the increase in plasma glucose and inhibited somatostatin expression induced by Q11206 , ( ii ) reduction of insulin prevalence was inhibited , while expression of p75 neurotrophin receptor ( p75NTR ) , pancreatic nerve growth factor ( P01138 ) , and P01138 -positive islet cell prevalence increased , ( iii ) there were no alterations in the severity of proliferated cell nuclear antigen positive or apoptotic beta cells in pancreatic islets , and ( iv ) pancreatic catalase , glutathione peroxidase , and superoxide dismutase activities significantly increased . In conclusion , these data suggest that exendin-4 might exert its actions through the P01138 /p75NTR system and decrease somatostatin expression . Effects of DB09045 on Thyroid C Cells and Serum P01258 in Male Monkeys . Glucagon-like peptide-1 ( P0C6A0 ) receptor agonists , used for the treatment of type 2 diabetes , have caused hyperplasia/neoplasia of thyroid C cells in rodent carcinogenicity studies . Studies in monkeys have not identified an effect of P43220 agonists on thyroid C cells ; however , group sizes were small . DB09045 is a once-weekly , long-acting human P43220 agonist recently approved in the United States and the European Union . The objective of this study was to determine whether dulaglutide altered C-cell mass in monkeys . Male cynomolgus monkeys ( 20 per group ) were sc injected with dulaglutide 8.15 mg/kg ( ∼500-fold maximum human plasma exposure ) or a vehicle control twice weekly for 52 weeks . Basal and calcium gluconate-stimulated serum calcitonin concentrations were obtained at 3 , 6 , 9 , and 12 months . Thyroid glands were weighed , fixed , and sectioned at 500-μm intervals . C-cell volumes were measured using an automated image analysis . C-cell proliferation was estimated using Ki67/calcitonin colabeling and cell counting . Administration of dulaglutide 8.15 mg/kg twice weekly for 52 weeks did not increase serum calcitonin in monkeys or affect thyroid weight , histology , C-cell proliferation , or absolute/relative C-cell volume . This study represents a comprehensive evaluation of the monkey thyroid C cells after dosing with a P43220 agonist , with a large group size , and measurement of multiple relevant parameters . The lack of effect of dulaglutide on C cells is consistent with other studies in monkeys using P43220 agonists and suggests that nonhuman primates are less sensitive than rodents to the induction of proliferative changes in thyroid C cells by P43220 agonists . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . Effect of the combination of metformin and fenofibrate on glucose homeostasis in diabetic Goto-Kakizaki rats . Metformin has been reported to increase the expression of the glucagon-like peptide-1 ( P0C6A0 ) receptor in pancreatic beta cells in a peroxisome proliferator-activated receptor ( Q07869 ) -α-dependent manner . We investigated whether a PPARα agonist , fenofibrate , exhibits an additive or synergistic effect on glucose metabolism , independent of its lipid-lowering effect , when added to metformin . Non-obese diabetic Goto-Kakizaki ( GK ) rats were divided into four groups and treated for 28 days with metformin , fenofibrate , metformin plus fenofibrate or vehicle . The random blood glucose levels , body weights , food intake and serum lipid profiles were not significantly different among the groups . After 4 weeks , metformin , but not fenofibrate , markedly reduced the blood glucose levels during oral glucose tolerance tests , and this effect was attenuated by adding fenofibrate . Metformin increased the expression of the P43220 in pancreatic islets , whereas fenofibrate did not . During the intraperitoneal glucose tolerance tests with the injection of a P0C6A0 analog , metformin and/or fenofibrate did not alter the insulin secretory responses . In conclusion , fenofibrate did not confer any beneficial effect on glucose homeostasis but reduced metformin 's glucose-lowering activity in GK rats , thus discouraging the addition of fenofibrate to metformin to improve glycemic control . DB09045 for the treatment of type 2 diabetes . DB09045 is a novel glucagon-like peptide 1 ( P0C6A0 ) receptor agonist with a unique structure that supports once-weekly dosing in patients with type 2 diabetes ( T2DM ) , most of whom have a big pill burden . It appears to be efficacious in reducing hemoglobin A1c ( HbA1c ) up to 1.59 % and promotes modest weight loss up to 3 kg with a low incidence of hypoglycemia and mild to moderate gastrointestinal adverse events . Convenient weekly dosing could improve compliance and help attain sustained glycemic goals . Addressing obesity is an integral part of T2DM management and weight loss may contribute to better glycemic and cardiovascular benefits . Results of ongoing clinical trials on cardiovascular safety are important to determine the risk-to-benefit ratio . As with any drug , patient selection and ongoing monitoring will be important . If approved , dulaglutide will be one of the first weekly P43220 agonist to be available in a ready-to-use pen device with an automatic injector . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . P43220 agonist may activate pancreatic stellate cells to induce rat pancreatic tissue lesion . OBJECTIVE : To explore the mechanism of P43220 agonist-induced rat pancreatic tissue lesion . METHODS : Thirty SD male rats were divided into three groups , namely P43220 agonist experimental group , diabetes-model experimental group and control group . Diabetes-model rats were induced by streptozotocin and high-sugar high-fat diet . P43220 agonist group and diabetes-model group were administered with P43220 agonist in dose 5 μg/kg each time , twice a day . After 10 weeks of treatment , the amount of matrix metalloproteinase ( MMP ) -2 and P14780 , and expression of α-smooth muscle actin ( α-SMA ) and type III collagen protein in pancreatic tissue were measured . RESULTS : The amount of P08253 and P14780 in P43220 agonist group and diabetes-model group were significantly higher than the control group . Compared with the P43220 agonist group , the diabetic model group had more severe pathological changes of pancreatic tissue interstitial edema , inflammatory cell infiltration , glandular atrophy and fibrosis , and significantly increased pancreatic tissue P08253 and P14780 levels , significantly increased α-SMA and collagen III-positive cell counts , all the differences were statistically significant . α-SMA and type III collagen were expressed in all parts of the lesions of P43220 agonist group and diabetes-model group . α-SMA can only be observed in the vessel wall in control group , however , in the other two groups , α-SMA can also be observed in pancreatic acinar cell interstitia , in addition to vessel wall . CONCLUSIONS : Long-term subcutaneous P43220 agonist injection may activate pancreatic stellate cells , causing the expression of α-SMA and collagen III and the amount of P08253 and P14780 in pancreatic acinar cell interstitial significantly increasing , and thus inducing chronic inflammatory change . Exendin-4 attenuates lipopolysaccharides induced inflammatory response but does not protects H9c2 cells from apoptosis . BACKGROUND : Glucagon-like peptide-1 ( P0C6A0 ) and its analogues are reported to exert wide-ranging cardiovascular actions in preclinical and clinical studies . We thus investigated whether the P43220 agonist , exendin-4 , has inhibitory effects on LPS-stimulated inflammatory response in cardiomyoblasts . METHODS : H9c2 cardiomyoblasts were exposed to LPS and treated with exendin-4 . Expressions of proinflammatory mediators were assessed using quantitative real-time PCR . Nuclear localization of NF-κB was examined using immunoblotting . mRNA expression of inducible nitric oxide synthase ( P35228 ) and nitric oxide ( NO ) production were evaluated by q PCR and NO assay . Furthermore , anti-apoptotic effect of exendin-4 in LPS-stimulated H9c2 cells was determined using qPCR and immunoblot . RESULTS : Exposure to LPS increased mRNA expressions of P01375 -α , P35354 and P14780 in H9c2 cells . It also caused increases in P35228 mRNA expression and NF-κB nuclear translocation . Exendin-4 dose-dependently downregulated mRNA levels of P01375 -α , P35354 and P14780 in LPS-stimulated H9c2 cells . It also reduced NF-κB nuclear translocation . Treatment with exendin-4 showed no effect on LPS-induced apoptosis in H9c2 cells . CONCLUSIONS : Exendin-4 exerts an effect on cardiomyoblast exposed to LPS by inhibiting mRNA expression of inflammatory mediators and suppressing NF-κB activation . These effects are consistent with some of the observed anti-inflammatory properties of exendin-4 , as well as its beneficial actions on the cardiovascular system . The contribution of serotonin P28335 and melanocortin-4 receptors to the satiety signaling of glucagon-like peptide 1 and liraglutide , a glucagon-like peptide 1 receptor agonist , in mice . Glucagon-like peptide 1 ( P0C6A0 ) , an insulinotropic gastrointestinal peptide produced mainly from intestinal endocrine L-cells , and liraglutide , a P43220 ( P43220 ) agonist , induce satiety . The serotonin P28335 receptor ( 5-HT2CR ) and melanoroctin-4 receptor ( P32245 ) are involved in the regulation of food intake . Here we show that systemic administration of P0C6A0 ( 50 and 200μg/kg ) -induced anorexia was blunted in mice with a 5HT2CR null mutation , and was attenuated in mice with a heterozygous P32245 mutation . On the other hand , systemic administration of liraglutide ( 50 and 100μg/kg ) suppressed food intake in mice lacking 5-HT2CR , mice with a heterozygous mutation of P32245 and wild-type mice matched for age . Moreover , once-daily consecutive intraperitoneal administration of liraglutide ( 100μg/kg ) over 3days significantly suppressed daily food intake and body weight in mice with a heterozygous mutation of P32245 as well as wild-type mice . These findings suggest that P0C6A0 and liraglutide induce anorexia via different central pathways . P0C6A0 treatment reduces endogenous insulin resistance via activation of central P0C6A0 receptors in mice fed a high-fat diet . Glucagon-like peptide-1 ( P0C6A0 ) improves insulin sensitivity in humans and rodents . It is currently unknown to what extent the ( metabolic ) effects of P0C6A0 treatment are mediated by central P0C6A0 receptors . We studied the impact of central P43220 ( P43220 ) antagonism on the metabolic effects of peripheral P0C6A0 administration in mice . High-fat-fed insulin-resistant C57Bl/6 mice were treated with continuous subcutaneous infusion of P0C6A0 or saline ( PBS ) for 2 wk , whereas the P43220 antagonist exendin-9 ( EX-9 ) and cerebrospinal fluid ( P04141 ) were simultaneously infused in the left lateral cerebral ventricle ( icv ) . DB09341 and glycerol turnover were determined during a hyperinsulinemic euglycemic clamp . VLDL-triglyceride ( VLDL-TG ) production was determined in hyperinsulinemic conditions . Our data show that the rate of glucose infusion necessary to maintain euglycemia was significantly increased by P0C6A0 . Simultaneous icv infusion of EX-9 diminished this effect by 62 % . The capacities of insulin to stimulate glucose disposal and inhibit glucose production were reinforced by P0C6A0 . Simultaneous icv infusion of EX-9 significantly diminished the latter effect . Central P43220 antagonism alone did not affect glucose metabolism . Also , P0C6A0 treatment reinforced the inhibitory action of insulin on VLDL-TG production . In conclusion , peripheral administration of P0C6A0 reinforces the ability of insulin to suppress endogenous glucose and VLDL-TG production ( but not lipolysis ) and boosts its capacity to stimulate glucose disposal in high-fat-fed C57Bl/6 mice . Activation of central GLP-1Rs contributes substantially to the inhibition of endogenous glucose production by P0C6A0 treatment in this animal model . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . Suppressive effects of glucagon-like peptide-1 on interferon-gamma-induced nitric oxide production in insulin-producing cells is mediated by inhibition of tumor necrosis factor-alpha production . During the development of Type 1 diabetes , inflammatory cytokines are known to induce the expression of inducible nitric oxide synthase ( P35228 ) in pancreatic islets , and subsequent production of nitric oxide ( NO ) contributes to beta cell destruction . Glucagon-like peptide-1 ( P0C6A0 ) has been shown to reduce cytokine-induced apoptosis of beta cells . In this study , we investigated whether P0C6A0 affects cytokine-induced NO production , resulting in the inhibition of beta-cell apoptosis . We treated MIN6N8a mouse beta cells with interferon ( IFN ) -gamma in the presence or absence of P0C6A0 and found that P01579 treatment induced P35228 mRNA expression and NO production , which was significantly inhibited by treatment with P0C6A0 . Blocking of P43220 signaling via the cyclic AMP and phosphatidylinositol 3-kinase pathway did not directly affect the suppressive effect of P0C6A0 on IFN- gamma-induced P35228 mRNA expression . Further studies revealed that P01579 induced the expression of P01375 mRNA and protein , which synergistically induced NO production , and P0C6A0 treatment inhibited this induction of P01375 . To examine whether the reduction of P01375 by P0C6A0 treatment plays a role in suppressing NO production , we treated MIN6N8a cells with P01579 in the presence of anti- P01375 neutralizing antibody and found that NO production was reduced . In addition , treatment of mouse islets with P0C6A0 inhibited the expression of P35228 and TNFmRNA . These results suggest that P0C6A0 inhibits P01579 -induced NO production by suppression of P01375 production . Polyamines and membrane transporters . In recent years , our understanding of the importance of membrane transporters ( MTs ) in the disposition of and response to drugs has increased significantly . MTs are proteins that regulate the transport of endogenous molecules and xenobiotics across the cell membrane . In mammals , two super-families have been identified : DB00171 -binding cassette ( DB01048 ) and solute carrier ( O00585 ) transporters . There is evidence that MTs might mediate polyamines ( PA ) transport . PA are ubiquitous polycations which are found in all living cells . In mammalian cells , three major PA are synthesised : putrescine , spermidine and spermine ; whilst the decarboxylated arginine ( agmatine ) is not produced by mammals but is synthesised by plants and bacteria . In addition , research in the PA field suggests that PA are transported into cells via a specific transporter , the polyamine transport system(s) ( Q03393 ) . Although the Q03393 has not been fully defined , there is evidence that some of the known MTs might be involved in PA transport . In this mini review , eight O00585 transporters will be reviewed and their potential to mediate PA transport in human cells discussed . These transporters are O15245 , O15244 , O75751 , Q96FL8 , P30825 , P08195 , SLC12A8A , and Q86VW1 . Preliminary data from our laboratory have revealed that O15245 might be involved in the PA uptake ; in addition to one member of ABC superfamily ( P08183 protein ) might also mediate the efflux of polyamine like molecules . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Cutting edge : peripheral neuropeptides attract immature and arrest mature blood-derived dendritic cells . Dendritic cells ( DC ) are highly motile and play a key role in mediating immune responses in various tissues and lymphatic organs . We investigated locomotion of mononuclear cell-derived DC at different maturation stages toward gradients of sensory neuropeptides in vitro . P01258 gene-related peptide , vasoactive intestinal polypeptide , secretin , and secretoneurin induced immature DC chemotaxis comparable to the potency of RANTES , whereas DB05875 and macrophage-inflammatory protein-3beta stimulated immature cell migration only slightly . Checkerboard analyses revealed a true chemotactic response induced by neuropeptides . Upon maturation of DC , neuropeptides inhibited spontaneous , macrophage-inflammatory protein-3beta- and O00585 -induced cell migration . Maturation-dependent changes in migratory behavior coincided with distinct neuropeptide-induced signal transduction in DC . Peripheral neuropeptides might guide immature DC to peripheral nerve fibers where high concentrations of these peptides can arrest the meanwhile matured cells . It seems that one function of sensory nerves is to fasten DC at sites of inflammation . Population pharmacokinetic study of memantine : effects of clinical and genetic factors . BACKGROUND AND OBJECTIVE : Memantine , a frequently prescribed anti-dementia drug , is mainly eliminated unchanged by the kidneys , partly via tubular secretion . Considerable inter-individual variability in plasma concentrations has been reported . We aimed to investigate clinical and genetic factors influencing memantine disposition . METHODS : A population pharmacokinetic study was performed including data from 108 patients recruited in a naturalistic setting . Patients were genotyped for common polymorphisms in renal cation transporters ( O15245 /2/5 , Q96FL8 , P08183 ) and nuclear receptors ( O75469 , Q14994 , RXR , Q07869 ) involved in transporter expression . RESULTS : The average clearance was 5.2 L/h with a 27 % inter-individual variability ( percentage coefficient of variation ) . Glomerular filtration rate ( p = 0.007 ) and sex ( p = 0.001 ) markedly influenced memantine clearance . O75469 rs1523130 was identified as the unique significant genetic covariate for memantine clearance ( p = 0.006 ) , with carriers of the O75469 rs1523130 CT/TT genotypes presenting a 16 % slower memantine elimination than carriers of the CC genotype . CONCLUSION : The better understanding of inter-individual variability of memantine disposition might be beneficial in the context of individual dose optimization . P01258 gene-related peptide is a potent inhibitor of DB05875 degradation . P01258 gene-related peptide ( P80511 ) was found to potently inhibit a DB05875 endopeptidase isolated from human P04141 . P80511 potentiated DB05875 irritant actions ; a possible mechanism is interaction for a common metabolic step . Somatostatin is another peptide capable of competing with DB05875 endopeptidase . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . DB09045 : the newest P43220 agonist for the management of type 2 diabetes . OBJECTIVE : To review the pharmacology , pharmacokinetics , safety , and efficacy of the glucagon-like peptide-1 receptor agonist ( P0C6A0 RA ) , dulaglutide , in the treatment of type 2 diabetes mellitus ( T2D ) . DATA SOURCES : A PubMed search was completed to identify publications from 1947 to October 2014 using the search terms dulaglutide and LY2189265 . References were reviewed to identify additional resources . STUDY SELECTION AND DATA EXTRACTION : Articles were included if they evaluated the pharmacology , pharmacokinetics , safety , or efficacy of dulaglutide . DATA SYNTHESIS : DB09045 reduces both glycosylated hemoglobin ( A1C ) and weight by stimulating insulin secretion and suppressing glucagon in a glucose-dependent manner , delaying gastric emptying , and promoting satiety . DB09045 consists of 2 P0C6A0 analogues that have been modified to make it a long-acting , once-weekly agent . DB09045 has been studied as monotherapy and in combination with metformin , glimepiride , pioglitazone , and insulin lispro . It has demonstrated superior A1C reduction compared with placebo , metformin , insulin glargine , sitagliptin , and twice-daily exenatide . It demonstrated noninferiority in A1C reduction to liraglutide . DB09045 changed A1C by -0.78 % to -1.51 % , and it changed weight by -0.35 kg to -3.03 kg . The most common adverse effects in clinical studies were nausea , vomiting , and diarrhea . CONCLUSIONS : DB09045 is the fifth P0C6A0 RA approved for T2D in the United States . It is an attractive option because it is dosed once-weekly , provides A1C lowering similar to liraglutide , weight reduction similar to exenatide , and has an adverse effect profile similar to exenatide and liraglutide . Primary angiosarcoma of the thyroid gland with recurrence diagnosed by fine needle aspiration : a case report . Angiosarcoma of the thyroid is a rare and aggressive primary malignant tumor of the thyroid originally reported in patients from the Swiss Alpine region . Diagnosis of this tumor rests mainly on characteristic histopathological features of a malignant vascular tumor supported by immunopositivity for vascular markers e.g. , CD31 , Factor VIII , and P28906 . Its cytological features , however , are not well-defined . We describe a case of primary angiosarcoma of the thyroid in a 48-year-old female , who presented with a rapidly enlarging neck mass associated with compressive symptoms . She had a history of hypothyroidism . The initial fine needle aspiration cytology of the neck mass was negative . She then underwent left hemithyroidectomy . Histologically , the tumor showed poorly differentiated malignant cells with eccentrically-placed nuclei , prominent nucleoli , and intracytoplasmic vacuoles admixed with mixed inflammatory cells . These showed immunopositivity for CD31 but were negative for P28906 , Factor VIII , CK5/6 , P15941 , Q15669 -1 , Thyroglobulin , P01258 , Melan A , and P22676 . A diagnosis of poorly differentiated malignant tumor consistent with angiosarcoma was made . The patient was treated with radiation therapy but developed recurrence of the tumor . Second aspiration cytology of the recurrent tumor yielded hypocellular smears containing singularly dispersed atypical cells having eccentrically-placed nuclei with prominent macronucleoli and intracytoplasmic vacuoles within a background of inflammatory cells , consistent with recurrent angiosarcoma . Chemotherapy was started but she succumbed to the disease 7 months after diagnosis . The cytological , histopathological , immunohistochemical findings , and the clinical course are discussed . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development .

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DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Role of presynaptic serotonergic receptors on the mechanism of action of P08908 and P28222 agonists on masculine sexual behaviour : physiological and pharmacological implications . In order to establish whether the P08908 or the 5HT1B agonists , 8-OH-DPAT or TFMPP , produce their facilitatory or inhibitory actions on masculine sexual behaviour via a mechanism involving : ( a ) the serotonin synthesis or release ; ( b ) the stimulation of presynaptic receptors , or ( c ) the stimulation of somatodendritic receptors , three series of experiments were performed . The administration of the serotonin synthesis inhibitor , p-chlorophenylalanine ( p- P15085 , 300 mg/kg x 3 days ) , facilitated sexual behaviour but does not interfere neither with the inhibitory nor with the facilitatory effects of TFMPP ( 0.5 mg/kg ) or 8-OH-DPAT ( 0.5 mg/kg ) , respectively . The icv or the intraraphé administration of the serotonergic neurotoxin , 5,7-dihydroxytryptamine ( 5,7- DB02901 ) , slightly stimulated masculine sexual behaviour and produced a decrease in serotonin and its metabolite levels . In lesioned animals TFMPP ( 0.5 mg/kg ) resulted in an inhibitory effect reflected as a prolongation of the ejaculation latency . The inhibitory effect of this drug on mounting behaviour was not observed in 5,7- DB02901 treated rats . In lesioned animals 8-OH-DPAT ( 0.5 mg/kg ) produced the same facilitatory effect . Present data indicate that serotonergic postsynaptic receptors mediate both the inhibitory and the facilitatory actions of TFMPP or 8-OH-DPAT in copulation . All data further support the idea that endogenous serotonin acts via the stimulation of P28222 receptors to induce its inhibitory effects on masculine sexual behaviour . Dopamine receptors and psychiatric drug treatment . The established antipsychotic drugs act mainly by antagonizing dopamine mediated synaptic transmission in the brain . Increase in the rate of production of dopamine metabolites as well as the firing rate of dopamine-containing neurons can be interpreted as compensatory responses to an interruption of synaptic transmission at dopamine nerve terminals . The demonstration of involvement of limbic and cortical mechanisms in the antipsychotic activity of neuroleptic drugs is far more difficult than the involvement of nigro-striatal and tubero-infundibular mechanisms in the neurological and neuroendocrine effects of these drugs . Application of radioreceptor techniques to dopamine research has supported the findings obtained by other neuropsychopharmacological research techniques , providing more direct evidence of dopamine receptor blockade by neuroleptic drugs . Further research is needed especially in studying the nature of the time-dependent adaptive changes at the receptor sites as well as the differences between the different dopamine projections and neural systems in the brain . The different subtypes of dopamine receptors in the brain , currently called D1 and D2 dopamine receptors , seem to be parallel , although in many respects independently-acting regulatory systems . P14416 -selective antagonists such as sulpiride seem to cause selective D2 receptor up-regulation . P01236 secretion seems to be regulated by D2 dopamine receptors . The exact physiological role of D1 dopamine receptors as well as the clinical consequences of selective D1 antagonism is not known . DB00391 and clozapine are examples of atypical neuroleptic compounds that have quite different profile of action , the former having strong and selective antidopaminergic action , the latter combining a number of non- dopaminergic mechanisms with rather slight effects on dopamine receptors. ( ABSTRACT TRUNCATED AT 250 WORDS ) P13569 mediated chloride secretion in the avian renal proximal tubule . In primary cell cultures of the avian ( Gallus gallus ) renal proximal tubule parathyroid hormone and DB02527 activation generate a Cl(-)-dependent short circuit current ( I(SC) ) response , consistent with net transepithelial Cl(-) secretion . In this study we investigated the expression and physiological function of the Na-K-2Cl ( NKCC ) transporter and P13569 chloride channel , both associated with Cl(-) secretion in a variety of tissues , in these proximal tubule cells . Using both RT-PCR and immunoblotting approaches , we showed that NKCC and P13569 are expressed , both in proximal tubule primary cultures and in a proximal tubule fraction of non-cultured ( native tissue ) fragments . We also used electrophysiological methods to assess the functional contribution of NKCC and P13569 to forskolin-activated I(SC) responses in filter grown cultured monolayers . DB00887 ( 10 μM ) , a specific blocker of NKCC , inhibited forskolin activated I(SC) by about 40 % , suggesting that basolateral uptake of Cl(-) is partially mediated by NKCC transport . In monolayers permeabilized on the basolateral side with nystatin , forskolin activated an apical Cl(-) conductance , manifested as bidirectional diffusion currents in the presence of oppositely directed Cl(-) gradients . Under these conditions the apical conductance appeared to show some bias towards apical-to-basolateral Cl(-) current . Two selective P13569 blockers , P13569 Inhibitor 172 and GlyH-101 ( both at 20 μM ) inhibited the forskolin activated diffusion currents by 38-68 % , with GlyH-101 having a greater effect . These data support the conclusion that avian renal proximal tubules utilize an apical P13569 Cl(-) channel to mediate DB02527 -activated Cl(-) secretion . Enhanced IgE allergic response to Aspergillus fumigatus in P13569 -/- mice . To gain insight into aberrant cytokine regulation in cystic fibrosis ( CF ) , we compared the phenotypic manifestations of allergen challenge in gut-corrected P13569 -deficient mice with background-matched C57Bl6 ( B6 ) mice . Aspergillus fumigatus ( Af ) antigen was used to mimic allergic bronchopulmonary aspergillosis , a peculiar hyper-IgE syndrome with a high prevalence in CF patients . P13569 -/- , C57BL/6 and FVB/NJ mice were sensitized with Af antigen by serial intraperitoneal injections . Control mice were mock sensitized with PBS . Challenges were performed by inhalation of Af antigen aerosol . After Af antigen challenge , histologic analysis showed goblet cell hyperplasia and lymphocytic infiltration in both strains . However , total serum IgE levels were markedly elevated in CF mice . Sensitized CF mice showed a five-fold greater IgE response to sensitization as compared with B6- and FVB-sensitized controls . Additional littermate controls to fully normalize for B6-FVB admixture in the strain background confirmed the role of P13569 mutation in the hyper-IgE syndrome . Cytokine mRNA levels of P05113 and GM- P04141 in the bronchoalveolar lavage ( BAL ) fluid , and BAL cell differentials indicated that P13569 mutation caused a shift from an P05113 -predominant to an P05112 -predominant cytokine profile . This system models a very specific type of airway inflammation in CF and could provide insights into pathogenesis and treatment of the disease . Role of chloride transport proteins in the vasorelaxant action of nitroprusside in isolated rat aorta . Chloride ions play a key role in smooth muscle contraction , but little is known concerning their role in smooth muscle relaxation . Here we investigated the effect of chloride transport inhibitors on the vasorelaxant responses to nitroprusside in isolated and endothelium-denuded rat aorta , precontracted with phenylephrine 1 muM . Incubation of aortic rings in NO(3)(-) media strongly potentiated the vasorelaxant responses to nitroprusside . DB00887 , DIDS ( 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid ) and acetazolamide strongly potentiated the vasorelaxant responses to nitroprusside ( by 70-100 % ) . EC(50) were 2.3+/-0.5 microM for bumetanide , 26+/-15 microM for DIDS and 510+/-118 microM for acetazolamide ( n=6 for condition ) . Niflumic acid , a selective inhibitor of ClCa ( calcium-activated chloride channels ) , potentiated nitroprusside relaxation to a similar extent as chloride transport inhibitors , in a non-additive manner . Zinc and nickel ions , both modestly potentiated nitroprusside vasorelaxation ( by 20-30 % ) . Cobaltum had negligible effect on nitroprusside vasorelaxation . P15085 ( p-chlorophenoxy-acetic acid ) , an inhibitor of volume-sensitive chloride channels ( ClC ) , slightly potentiated nitroprusside vasorelaxation ( by 15 % ) , and the cystic fibrosis transmembrane conductance regulator ( P13569 ) chloride channel inhibitors P13569 (inh)172 ( 5-[(4-Carboxyphenyl)methylene]-2-thioxo-3- [ (3-trifluoromethyl)phenyl-4-thiazolidinone ) , DPC ( diphenylamine-2,2'-dicarboxylic acid ) and glibenclamide were without significant effect . In conclusion , inhibition of chloride transport proteins strongly potentiates the vasorelaxant responses to nitroprusside in isolated rat aorta . This effect seems mediated by chloride depletion and inhibition of a chloride channel activated by both , calcium and cyclic GMP ( cGMP ) . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Increased epithelial permeability is the primary cause for bicarbonate loss in inflamed murine colon . BACKGROUND : Bicarbonate loss into the lumen occurs during intestinal inflammation in different species . However , candidate pathways like P13569 or P40879 are inhibited in the inflamed gut . This study addressed the question whether and how inflammation-associated increased intestinal permeability may result in epithelial HCO(3)(-) loss . METHODS : Murine proximal colon was studied because it does not express functional P40879 but is inflamed in the tumor necrosis factor α overexpressing mouse model ( P01375 (ΔARE) ) . DB01174 alkalization , (3)H-mannitol fluxes , impedance spectroscopy , and dilution potentials were measured in Ussing chambers , whereas expression and localization of tight junction-associated proteins were analyzed by Western blots and immunohistochemistry . RESULTS : DB01174 alkalization rates and (3)H-mannitol fluxes were increased in P01375 (+/ΔARE) proximal colon , whereas forskolin-stimulated I(sc) was not altered . Epithelial resistance was reduced , but subepithelial resistance increased . The epithelial lining was intact , and enterocyte apoptosis rate was not increased despite massively increased Th1 cytokine levels and lymphoplasmacellular infiltration . Measurement of dilution potentials suggested a loss of cation selectivity with increased anion permeability . Western analysis revealed a downregulation of occludin expression and an upregulation of both claudin-2 and claudin-5 , with no change in ZO-1 , P12830 , claudin-4 , and claudin-8 . Immunohistochemistry suggested correct occludin localization but reduced tight junction density in P01375 (+/ΔARE) surface epithelium . CONCLUSIONS : Inflammation during P01375 -α overexpression leads to increased epithelial permeability in murine proximal colon , decreased tight junctional cation selectivity , and increased HCO(3)(-) loss into the lumen . Inflammation-associated colonic HCO(3)(-) loss may occur through leaky tight junctions rather than through HCO(3)(-) secreting ion transporters . Microelectrode measurements of the effects of basolateral adenosine in polarized human intestinal epithelial cells in culture . Activation of the basolateral receptor for adenosine in HT-29cl.19A cells , by 100 microM adenosine , increased the equivalent short-circuit current ( DeltaIsc= 24+/-2 microA/cm2 ) , depolarized the intracellular potential ( DeltaVa= 26+/-2 mV ) and decreased the fractional apical membrane resistance ( DeltafRa=-0.48 ) . The changes in all parameters reached their peak values simultaneously . This suggests that the primary action of the adenosine-activated pathway is on only one membrane . DB00887 inhibited the transepithelial response and repolarized the cell potential . After preincubation with 100 microM forskolin , application of 300 microM adenosine caused a significant further change in Va , Isc , the transepithelial potential ( Vt ) and fRa . Together with the results from ion-replacement studies , the observations indicate that adenosine activates channels other than the cystic fibrosis transmembrane conductance regulator ( P13569 ) . The rank order of potencies of adenosine and adenosine analogues implies that the receptor is of the A2 subtype . Preincubation with 4-bromophenacyl bromide ( 4-BPB ) inhibited the effect of an adenosine analogue by 50 % , indicating that activation of phospholipase A2 may be involved in the adenosine-induced response . DB02527 inhibition of murine intestinal Na/H exchange requires P13569 -mediated cell shrinkage of villus epithelium . BACKGROUND AND AIMS : Unlike the intestine of normal subjects , small-intestinal epithelia of cystic fibrosis patients and cystic fibrosis transmembrane conductance regulator protein-null ( P13569 (-) ) mice do not respond to stimulation of intracellular cyclic adenosine monophosphate with inhibition of electroneutral NaCl absorption . Because P13569 -mediated anion secretion has been associated with changes in crypt cell volume , we hypothesized that P13569 -mediated cell volume reduction in villus epithelium is required for intracellular cyclic adenosine monophosphate inhibition of Na(+)/H(+) exchanger ( primarily P48764 ) activity in the proximal small intestine . METHODS : Transepithelial (22)Na flux across the jejuna of P13569 (+) , P13569 (-) , the basolateral membrane Na(+)/K(+)/2Cl(-) co-transporter protein P55011 (+) , and P55011 (-) mice were correlated with changes in epithelial cell volume of the midvillus region . RESULTS : Stimulation of intracellular cyclic adenosine monophosphate resulted in cessation of Na(+)/H(+) exchanger-mediated Na(+) absorption ( J(ms)(NHE) ) in P13569 (+) jejunum but had no effect on J(ms)(NHE) across P13569 (-) jejunum . Cell volume indices indicated an approximately 30 % volume reduction of villus epithelial cells in P13569 (+) jejunum but no changes in P13569 (-) epithelium after intracellular cyclic adenosine monophosphate stimulation . In contrast , cell shrinkage induced by hypertonic medium inhibited J(ms)(NHE) in both P13569 (+) and P13569 (-) mice . DB00887 treatment to inhibit Cl(-) secretion by blockade of the Na(+)/K(+)/2Cl(-) co-transporter , P55011 , of stimulated P13569 (+) jejunum prevented maximal volume reduction of villus epithelium and recovered approximately 40 % of J(ms)(NHE) . Likewise , J(ms)(NHE) and cell volume were unaffected by intracellular cyclic adenosine monophosphate stimulation in P55011 (-) jejuna . CONCLUSIONS : These findings show a previously unrecognized role of functional P13569 expressed in villus epithelium : regulation of P48764 -mediated Na(+) absorption by alteration of epithelial cell volume . Methoxsalen stimulates electrogenic Cl- secretion in the mouse jejunum . We used the short-circuit current ( I(sc) ) and patch-clamp techniques to investigate the effects of methoxsalen ( MTX ) on the electrogenic Cl- secretion of the mouse jejunum . MTX stimulated a sustained increase in Isc that was dose dependent . DB00887 inhibited MTX-stimulated Isc in a dose-dependent manner consistent with activation of Cl- secretion . MTX failed to stimulate I(sc) following maximal activation of the DB02527 pathway by forskolin , but did increase Isc after a submaximal dose of forskolin . DB01016 , a blocker of the cystic fibrosis transmembrane conductance regulator ( P13569 ) , reduced the MTX-stimulated increase of Isc by 59 +/- 6 % . The DB02527 -dependent K+ channel blocker 293B did not alter the MTX-activated I(sc) ; however , clotrimazole , an intermediate Ca2(+)-activated K+ channel ( IK(Ca) ) blocker , reduced the MTX-stimulated I(sc) . MTX did not alter Na(+)-glucose cotransport across the mouse jejunum . In cell-attached membrane patches , MTX increased the open probability of the basolateral IK(Ca) channel of isolated crypts . These data suggest that the P13569 and IK(Ca) channels participate in the MTX-activated , sustained Cl- secretory response of the mouse jejunum . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Melatonin receptor potentiation of cyclic AMP and the cystic fibrosis transmembrane conductance regulator ion channel . We have used the cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channel as a model system to study the DB02527 signal transduction pathways coupled to the Xenopus melatonin receptor . During forskolin ( Fsk ) stimulation , melatonin reduced the amplitude of the P13569 currents in oocytes injected with in vitro transcribed cRNAs for the Xenopus melatonin receptor and P13569 . Pertussis toxin ( Ptx ) treatment eliminated melatonin inhibition of Fsk stimulated P13569 currents . In oocytes injected with cRNA for melatonin receptors , serotonin receptors ( P34969 ) , and P13569 Cl- channels , application of melatonin together with serotonin ( 5-HT ) activated an additional inward current showing potentiation of adenylyl cyclases by melatonin receptors . Subthreshold activation of P34969 receptors was sufficient and necessary to permit activation of P13569 channels by melatonin . Preexposure to melatonin desensitized the melatonin receptor mediated response . Therefore , based on this model system , the effects of melatonin in vivo could be either positive or negative modulation of other neuronal inputs , depending on the mode of adenylyl cyclase stimulation . DB00887 blocks P13569 GCl in the native sweat duct . DB00887 is well known for its ability to inhibit the nonconductive Na+-K+-2Cl- cotransporter . We were surprised in preliminary studies to find that bumetanide in the contraluminal bath also inhibited NaCl absorption in the human sweat duct , which is apparently poor in cotransporter activity . Inhibition was accompanied by a marked decrease in the transepithelial electrical conductance . Because the cystic fibrosis transmembrane conductance regulator ( P13569 ) Cl- channel is richly expressed in the sweat duct , we asked whether bumetanide acts by blocking this anion channel . We found that bumetanide 1 ) significantly increased whole cell input impedance , 2 ) hyperpolarized transepithelial and basolateral membrane potentials , 3 ) depolarized apical membrane potential , 4 ) increased the ratio of apical-to-basolateral membrane resistance , and 5 ) decreased transepithelial Cl- conductance ( GCl ) . These results indicate that bumetanide inhibits P13569 GCl in both cell membranes of this epithelium . We excluded bumetanide interference with the protein kinase A phosphorylation activation process by " irreversibly " phosphorylating P13569 [ by using adenosine 5'-O-(3-thiotriphosphate) in the presence of a phosphatase inhibition cocktail ] before bumetanide application . We then activated P13569 GCl by adding 5 mM DB00171 . DB00887 in the cytoplasmic bath ( 10(-3) M ) inhibited approximately 71 % of this DB00171 -activated P13569 GCl , indicating possible direct inhibition of P13569 GCl . We conclude that bumetanide inhibits P13569 GCl in apical and basolateral membranes independent of phosphorylation . The results also suggest that > 10(-5) M bumetanide can not be used to specifically block the Na+-K+-2Cl- cotransporter . Effects of salinity and prolactin on gene transcript levels of ion transporters , ion pumps and prolactin receptors in Mozambique tilapia intestine . Euryhaline teleosts are faced with significant challenges during changes in salinity . Osmoregulatory responses to salinity changes are mediated through the neuroendocrine system which directs osmoregulatory tissues to modulate ion transport . P01236 ( PRL ) plays a major role in freshwater ( FW ) osmoregulation by promoting ion uptake in osmoregulatory tissues , including intestine . We measured mRNA expression of ion pumps , Na(+)/K(+)-ATPase α3-subunit ( NKAα3 ) and vacuolar type H(+)-ATPase A-subunit ( V-ATPase A-subunit ) ; ion transporters/channels , Na(+)/K(+)/2Cl(-) co-transporter ( Q13621 ) and cystic fibrosis transmembrane conductance regulator ( P13569 ) ; and the two PRL receptors , PRLR1 and PRLR2 in eleven intestinal segments of Mozambique tilapia ( Oreochromis mossambicus ) acclimated to FW or seawater ( SW ) . Gene expression levels of NKAα3 , V-ATPase A-subunit , and Q13621 were generally lower in middle segments of the intestine , whereas P13569 mRNA was most highly expressed in anterior intestine of FW-fish . In both FW- and SW-acclimated fish , PRLR1 was most highly expressed in the terminal segment of the intestine , whereas PRLR2 was generally most highly expressed in anterior intestinal segments . While Q13621 , NKAα3 and PRLR2 mRNA expression was higher in the intestinal segments of SW-acclimated fish , P13569 mRNA expression was higher in FW-fish ; PRLR1 and V-ATPase A-subunit mRNA expression was similar between FW- and SW-acclimated fish . Next , we characterized the effects of hypophysectomy ( Hx ) and PRL replacement on the expression of intestinal transcripts . Hypophysectomy reduced both Q13621 and P13569 expression in particular intestinal segments ; however , only Q13621 expression was restored by PRL replacement . Together , these findings describe how both acclimation salinity and PRL impact transcript levels of effectors of ion transport in tilapia intestine . Initial interrogation , confirmation and fine mapping of modifying genes : P40763 , P01584 and P15260 determine cystic fibrosis disease manifestation . We have used a stepwise approach consisting of initial interrogation , confirmation and fine mapping to analyze P40763 , P01584 and P15260 as modifiers of cystic fibrosis disease building upon the data and sample collection of the European Cystic Fibrosis Twin and Sibling Study . We have observed direct correlation between the length of the intronic microsatellite STAT3Sat to P40763 expression levels among F508del- P13569 homozygous patients ( P=0.0075 ) , and an association of longer STAT3Sat-alleles with the presence of P13569 -mediated residual chloride secretion ( P=0.0031 ) , measured as the manifestation of the CF basic defect in intestinal tissue . Both , family-based analysis by P04053 and case-reference comparison identified consistently the same intragenic P01584 haplotype as a risk variant ( P(raw)=0.055 for P04053 , P(raw) < 0.3 for case-reference comparison ) . Using haplotype-guided hierarchical fine mapping , we have identified two single nucleotide exchanges for which concordant and discordant sibling pairs differ at a 7 kb-spanning core haplotype in P15260 ( P(raw)=0.0113 ) . Taken together , our findings imply that immunorelevant pathways and ion secretion , dominated by P13569 in intestinal and respiratory epithelium , merge at the level of the epithelial cell to integrate the signaling of cytokines due to innate and acquired immune defense . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Protein disulphide isomerase is required for signal peptide peptidase-mediated protein degradation . The human cytomegalovirus glycoprotein US2 induces dislocation of MHC class I heavy chains from the endoplasmic reticulum ( ER ) into the cytosol and targets them for proteasomal degradation . Q8TCT9 ( Q8TCT9 ) has been shown to be integral for US2-induced dislocation of MHC class I heavy chains although its mechanism of action remains poorly understood . Here , we show that knockdown of protein disulphide isomerase ( P07237 ) by RNA-mediated interference inhibited the degradation of MHC class I molecules catalysed by US2 but not by its functional homolog US11 . Overexpression of the substrate-binding mutant of P07237 , but not the catalytically inactive mutant , dominant-negatively inhibited US2-mediated dislocation of MHC class I molecules by preventing their release from US2 . Furthermore , P07237 associated with Q8TCT9 independently of US2 and knockdown of P07237 inhibited Q8TCT9 -mediated degradation of CD3delta but not Q9BUN8 -dependent degradation of P13569 DeltaF508 . Together , our data suggest that P07237 is a component of the Q8TCT9 -mediated ER-associated degradation machinery . DCEBIO stimulates Cl- secretion in the mouse jejunum . We investigated the effects of 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one(DCEBIO) on the Cl- secretory response of the mouse jejunum using the Ussing short-circuit current ( Isc ) technique . DCEBIO stimulated a concentration-dependent , sustained increase in Isc ( EC50 41 +/- 1 microM ) . Pretreating tissues with 0.25 microM forskolin reduced the concentration-dependent increase in Isc by DCEBIO and increased the EC50 ( 53 +/- 5 microM ) . DB00887 blocked ( 82 +/- 5 % ) the DCEBIO-stimulated Isc consistent with Cl- secretion . DCEBIO was a more potent stimulator of Cl- secretion than its parent molecule , 1-ethyl-2-benzimidazolinone . DB01016 or P16860 reduced the DCEBIO-stimulated Isc by > 80 % indicating the participation of P13569 in the DCEBIO-stimulated Isc response . Clotrimazole reduced DCEBIO-stimulated Isc by 67 +/- 15 % , suggesting the participation of the intermediate conductance Ca2+-activated K+ channel ( IKCa ) in the DCEBIO-activated Isc response . In the presence of maximum forskolin ( 10 microM ) , the DCEBIO response was reduced and biphasic , reaching a peak response of the change in Isc of 43 +/- 5 microA/cm2 and then falling to a steady-state response of 17 +/- 10 microA/cm2 compared with DCEBIO control tissues ( 61 +/- 6 microA/cm2 ) . The forskolin-stimulated Isc in the presence of DCEBIO was reduced compared with forskolin control tissues . Similar results were observed with DCEBIO and 8-BrcAMP where adenylate cyclase was bypassed . H89 , a PKA inhibitor , reduced the DCEBIO-activated Isc , providing evidence that DCEBIO increased Cl- secretion via a DB02527 /PKA-dependent manner . These data suggest that DCEBIO stimulates Cl- secretion of the mouse jejunum and that DCEBIO targets components of the Cl- secretory mechanism . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . P05231 and P10145 increase the expression of glycosyltransferases and sulfotransferases involved in the biosynthesis of sialylated and/or sulfated Lewisx epitopes in the human bronchial mucosa . Bronchial mucins from patients suffering from CF ( cystic fibrosis ) exhibit glycosylation alterations , especially increased amounts of the sialyl-Lewis(x) ( NeuAcalpha2-3Galbeta1-4[Fucalpha1-3]GlcNAc-R ) and 6-sulfo-sialyl-Lewis(x) ( NeuAcalpha2-3Galbeta1-4[Fucalpha1-3][SO(3)H-6]GlcNAc-R ) terminal structures . These epitopes are preferential receptors for Pseudomonas aeruginosa , the bacteria responsible for the chronicity of airway infection and involved in the morbidity and early death of CF patients . However , these glycosylation changes can not be directly linked to defects in P13569 ( CF transmembrane conductance regulator ) gene expression since cells that secrete airway mucins express no or very low amounts of the protein . Several studies have shown that inflammation may affect glycosylation and sulfation of various glycoproteins , including mucins . In the present study , we show that incubation of macroscopically healthy fragments of human bronchial mucosa with P05231 ( interleukin-6 ) or P10145 results in a significant increase in the expression of alpha1,3/4-fucosyltransferases [ Q495W5 ( fucosyltransferase 11 gene ) and P21217 ] , alpha2-6- and alpha2,3-sialyltransferases [ Q9Y274 ( alpha2,3-sialyltransferase 6 gene ) and Q96JF0 ( alpha2,6-sialyltransferase 2 gene ) ] and GlcNAc-6-O-sulfotransferases [ Q8NCG5 ( carbohydrate sulfotransferase 4 gene ) and Q9GZX3 ] mRNA . In parallel , the amounts of sialyl-Lewis(x) and 6-sulfo-sialyl-Lewis(x) epitopes at the periphery of high-molecular-mass proteins , including Q99102 , were also increased . In conclusion , our results indicate that P05231 and -8 may contribute to the increased levels of sialyl-Lewis(x) and 6-sulfo-sialyl-Lewis(x) epitopes on human airway mucins from patients with CF . DB01645 stimulates electrogenic Cl(-) secretion in mouse jejunum . We used the short-circuit current ( I(sc) ) technique to investigate the effects of the isoflavone genistein on the electrogenic Cl(-) secretion of the mouse jejunum . DB01645 stimulated a sustained increase in I(sc) that was dose dependent . DB00887 inhibited 76 +/- 5 % of the genistein-stimulated I(sc) consistent with activation of Cl(-) secretion . DB01645 failed to stimulate I(sc) following maximal activation of the DB02527 pathway by forskolin . In addition , forskolin had a reduced effect on I(sc) of the mouse jejunum in the presence of genistein . DB01016 , a blocker of P13569 , eliminated the genistein-stimulated increase of I(sc) and reduced the forskolin-activated I(sc) . Clotrimazole , a Ca(2+)-activated K(+) channel blocker , failed to reduce the genistein-stimulated I(sc) . Vanadate , a blocker of tyrosine-dependent phosphatases , reduced the genistein-activated I(sc) . Tyrphostin A23 , a tyrosine kinase inhibitor , reduced basal I(sc) , after which genistein failed to stimulate I(sc) . These data suggest that genistein activated a sustained Cl(-) secretory response of the mouse jejunum and that the effect of genistein was via a tyrosine-dependent phosphorylation pathway . A DNA hypermethylation profile reveals new potential biomarkers for prostate cancer diagnosis and prognosis . BACKGROUND : DNA hypermethylation has emerged as a novel molecular biomarker for the evaluation of prostate cancer diagnosis and prognosis . Defining the specific gene hypermethylation profile for prostate cancer could involve groups of genes that specifically discriminate patients with indolent and aggressive tumors . METHODS : Genome-wide methylation analysis was performed on 83 tumor and 10 normal prostate samples using the GoldenGate Methylation Cancer Panel I ( Illumina , Inc. ) . All clinical stages of disease were considered . RESULTS : We found 41 genes hypermethylated in more than 20 % of the tumors analyzed ( P < 0.01 ) . Of these , we newly identified P28161 and P01210 as being genes that are hypermethylated in prostate cancer and that were simultaneously methylated in 40.9 % of the tumors analyzed . We also identified panels of genes that are more frequently methylated in tumor samples with clinico-pathological indicators of poor prognosis : a high Gleason score , elevated Ki-67 , and advanced disease . Of these , we found simultaneous hypermethylation of P13569 and P28222 to be common in patients with a high Gleason score and high Ki-67 levels ; this might indicate the population at higher risk of therapeutic failure . The DNA hypermethylation profile was associated with cancer-specific mortality ( log-rank test , P = 0.007 ) and biochemical recurrence-free survival ( log-rank test , P = 0.0008 ) . CONCLUSIONS : Our findings strongly indicate that epigenetic silencing of P28161 and P01210 is a common event in prostate cancer that could be used as a molecular marker for prostate cancer diagnosis . In addition , simultaneous P28222 and P13569 hypermethylation could help discriminate aggressive from indolent prostate tumors . P13569 modulates neurosecretory function in pulmonary neuroendocrine cell-related tumor cell line models . The pulmonary neuroendocrine cell ( PNEC ) system consists of solitary cells and distinctive cell clusters termed neuroepithelial bodies ( P20929 ) localized in the airway epithelium . PNEC/ P20929 express a variety of bioactive substances , including amine ( serotonin , 5HT ) and neuropeptides . We have previously shown that P20929 cells are O(2) sensors expressing nicotinamide adenine diphosphate oxidase complex and O(2) sensitive K(+) channel . Recently , we demonstrated expression of functional cystic fibrosis transmembrane conductance regulator ( P13569 ) and Cl(-) conductances in P20929 cells of rabbit neonatal lung . Because PNEC/ P20929 are sparsely distributed and difficult to study in native lung , we investigated small-cell lung carcinoma ( SCLC ) and carcinoid tumor cell lines ( tumor counterparts of normal PNEC/ P20929 ) as models for PNEC/ P20929 . SCLC ( H146 , H345 ) and carcinoid ( H727 ) cell lines express neuroendocrine cell markers , including chromogranin A , neural cell adhesion molecule ( N- P62158 ) , 5HT , and tryptophan hydroxylase . We report that H146 , H345 , and H727 express P13569 messenger RNA ( reverse transcription polymerase chain reaction ) and protein ( immunoblotting ) and possess functional P13569 Cl(-) conductance , demonstrated by an iodide efflux assay inhibitable by transfection with antisense P13569 . Using an immunoassay to quantitate 5HT secretion , we also show that downregulation of P13569 abolishes hypoxia-induced 5HT release , and reduces secretory response to high potassium . Our findings suggest that P13569 may modulate neurosecretory activity of PNEC/ P20929 possessing O(2) sensor function . We propose that these tumor cell lines may be useful models for investigating the role of P13569 in PNEC/ P20929 functions in health and disease .

MH_train_29

MH_train_29

MH_train_29

interacts_with DB08916?

multiple_choice

DB08916 enhances the efficacy of conventional chemotherapeutic agents by eradicating cancer stem-like cells . Cancer stem cells ( CSC ) have garnered significant attention as a therapeutic focus , based on evidence that they may represent an etiologic root of treatment-resistant cells . Indeed , expression of the multidrug resistance protein DB00171 -binding cassette subfamily G member 2 ( Q9UNQ0 ) confers chemoresistance to CSCs , where it serves as a potential biomarker and therapeutic target . Here , we show that afatinib , a small-molecule inhibitor of the tyrosine kinases P00533 , P04626 , and Q15303 , preferentially eliminated side population cells with CSC character , in both cell lines and patient-derived leukemia cells , by decreasing Q9UNQ0 expression . In these cells , afatinib also acted in parallel to suppress self-renewal capacity and tumorigenicity . Combining afatinib with the DNA-damaging drug topotecan enhanced the antitumor effect of topotecan in vitro and in vivo . Mechanistic investigations suggested that Q9UNQ0 suppression by afatinib did not proceed by proteolysis through the ubiquitin-dependent proteosome , lysosome , or calpain . Instead , we found that afatinib increased DNA methyltransferase activity , thereby leading to methylation of the Q9UNQ0 promoter and to a decrease in Q9UNQ0 message level . Taken together , our results advocate the use of afatinib in combination with conventional chemotherapeutic drugs to improve efficacy by improving CSC eradication . DB08916 prolongs survival compared with gefitinib in an epidermal growth factor receptor-driven lung cancer model . An irreversible ErbB family blocker is expected to inhibit tumors with activating epidermal growth factor receptor ( P00533 ) mutations more strongly than reversible P00533 tyrosine kinase inhibitors and to overcome acquired resistance to the T790M secondary mutation . Eleven-week-old transgenic mice with Egfr exon 19 deletion mutation were treated with afatinib , gefitinib , or vehicle for 4 weeks . All mice were sacrificed at 15 weeks of age , and the number of superficial left lung tumors with a long axis exceeding 1 mm was counted . The afatinib-treated group had significantly fewer tumors than the vehicle group ( P < 0.01 ) and tended to have fewer tumors than the gefitinib-treated group ( P = 0.06 ) . Pathologically , gefitinib-treated mice had clearer , more nodular tumors than afatinib-treated mice . Immunoblotting showed that afatinib suppressed not only pEGFR but also pHER2 , and induced apoptosis for longer periods than gefitinib . Subsequently , when each drug was administered 5 days per week until death , afatinib significantly enhanced mouse survival compared with gefitinib ( median survival time : 456 days vs. 376.5 days ; log-rank test , P < 0.01 ) . Finally , the combination of afatinib with bevacizumab was found to be superior to either drug alone in exon 19 deletion/T790M and L858R/T790M xenograft tumors . Overall , afatinib was more potent than gefitinib in tumors harboring an exon 19 deletion mutation , and the combination of afatinib with bevacizumab efficiently suppressed tumors harboring the T790M secondary mutation . [ Retrospective Analysis of the DB08916 Clinical Pathway during the 28-Day Introductory Period-The Japanese Style of Collaborative Drug Therapy Management(J-CDTM) ] . DB08916 is a newly approved second-generation epidermal growth factor receptor-tyrosine kinase inhibito r( P00533 -TKI) . DB08916 has been shown to prolongthe overall survival of patients with non-small cell lungcancer ( NSCLC ) with P00533 mutations compared with the standard chemotherapy . However , Grade 3 or 4 toxicities , includingdiarrhea , rash , paronychia , and stomatitis , have been observed more frequently in patients treated with afatinib than in those treated with first-generation P00533 -TKIs . Accordingly , our institution developed an afatinib clinical pathway ( the afatinib pathway ) , which was designed by certified nurses , medical physicians , and certified pharmacists , with the goal of reducing the severity of diarrhea and rash that occur most frequently duringthe 28-day introductory period of afatinib treatment . Between May and October 2014 , afatinib was administered accordingto the afatinib pathway to 14 patients with NSCLC and P00533 mutations . Of these patients , only one ( 7.1 % ) experienced Grade 3 diarrhea . No other patient experienced Grade 3 or 4 toxicity . The afatinib pathway was effective in reducingthe severities of the diarrhea and rash duringthe 28-day introductory period of the afatinib treatment . Our implementation of the afatinib pathway could be considered the Japanese style of collaborative drugtherapy management ( J-CDTM ) . Targeted therapies : DB08916 -- new therapy option for P00533 -mutant lung cancer . YAP modifies cancer cell sensitivity to P00533 and survivin inhibitors and is negatively regulated by the non-receptor type protein tyrosine phosphatase 14 . The Yes-associated protein ( YAP ) is a transcriptional factor involved in tissue development and tumorigenesis . Although YAP has been recognized as a key element of the Hippo signaling pathway , the mechanisms that regulate YAP activities remain to be fully characterized . In this study , we demonstrate that the non-receptor type protein tyrosine phosphatase 14 ( Q15678 ) functions as a negative regulator of YAP . We show that YAP forms a protein complex with Q15678 through the WW domains of YAP and the PPXY motifs of Q15678 . In addition , Q15678 inhibits YAP-mediated transcriptional activities . Knockdown of YAP sensitizes cancer cells to various anti-cancer agents , such as cisplatin , the P00533 tyrosine kinase inhibitor erlotinib and the small-molecule antagonist of survivin , P28222 . YAP-targeted modalities may be used in combination with other cancer drugs to achieve maximal therapeutic effects . Clinical and comparative utility of afatinib in non-small cell lung cancer . The first targeted agents approved for non-small cell lung cancer ( NSCLC ) treatment , the epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors ( TKIs ) gefitinib and erlotinib , have an impressive activity in the presence of activating mutations of the P00533 gene . However , all patients develop acquired resistance principally through secondary mutations ( T790M ) , P04626 amplification , MET amplification , and other molecular aberrations . An attempt to overcome P00533 TKI resistance has been through the development of irreversible blockers . DB08916 is an irreversible inhibitor of the tyrosine kinase activity of all members of the HER family . The pharmacologic properties of afatinib ( formation of covalent bonds , inhibition of other family members , and in vitro and in vivo activity on T790M mutation positive tumors ) made this drug particularly appealing to study in clinic . Therefore , an intense program of clinical research ( LUX-Lung program ) was started and clinical results have shown very encouraging activity profiles in patients harboring P00533 activating mutations and in those with acquired resistance to reversible TKIs . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . Potential of afatinib in the treatment of patients with P04626 -positive breast cancer . In the absence of treatment , overexpression of the human epidermal growth factor receptor 2 ( P04626 ) predicts a poor prognosis in breast cancer . In the last decade , monoclonal antibodies and small molecule tyrosine kinase inhibitors have significantly improved the outcome of P04626 -positive breast cancer patients . However , tumor resistance and toxicities often limit the use of these therapies . For this reason , there is a compelling need for further investigation of new targeted therapies , such as afatinib , an oral irreversible pan inhibitor of the epidermal growth factor receptor ( P00533 ) family . This compound covalently interacts with tyrosine kinase domains , which are deeply involved in signal transduction leading to cell proliferation and protection from apoptosis . DB08916 has been studied in several Phase I clinical trials in advanced solid tumors . These trials have shown encouraging clinical activity and manageable side effects when afatinib is used either as a single agent or in combination with chemotherapy , with cutaneous adverse events and diarrhea being the most frequently observed toxicities . This review will focus on afatinib 's clinical activity and will discuss ongoing clinical studies in P04626 -positive breast cancer patients . In the scenario of the different P04626 -targeted therapies , it will be important to define the best specific clinical and " molecular " setting for afatinib use , trying to identify predictors of resistance and response . Moreover , afatinib , which has the ability to cross the blood-brain barrier , could play a role in patients with brain metastases from breast cancer . DB08916 : A first-line treatment for selected patients with metastatic non-small-cell lung cancer . PURPOSE : The pharmacology , pharmacokinetics , clinical efficacy , safety , adverse effects , dosage and administration , and role in therapy of afatinib in the management of non-small-cell lung cancer ( NSCLC ) are reviewed . SUMMARY : DB08916 ( Gilotrif , Boehringer Ingelheim ) is a novel oral tyrosine kinase inhibitor ( TKI ) recently approved for the first-line treatment of patients with NSCLC whose tumors are driven by activating mutations of genes coding for epidermal growth factor receptor ( P00533 ) . DB08916 is also an inhibitor of a specific P00533 mutation ( T790M ) that causes resistance to first-generation P00533 -targeted TKIs in about half of patients receiving those drugs . The recommended dosage is 40 mg once daily . In a Phase III trial completed last year , patients with P00533 -mutated NSCLC who were treated with afatinib had a twofold higher response rate than those receiving standard combination chemotherapy ( 56 % versus 23 % ) and significantly longer progression-free survival ( 11.0 months versus 5.6 months ) . Other studies indicated that afatinib may offer advantages over standard chemotherapy for NSCLC in terms of enhanced symptom control and quality of life and is modestly effective in cases involving EGFRT790M-related acquired resistance to the TKIs erlotinib and gefitinib . Among clinical trial participants , afatinib was generally well tolerated , with the most common grade I or II adverse events being diarrhea and rash or acne ; grade III or IV events were infrequent . CONCLUSION : DB08916 is a novel TKI that is efficacious and well tolerated in patients with NSCLC associated with activating P00533 mutations , including cases involving the T790M resistance mutation . It has possible applications in other P00533 mutation- positive cancers . DB08916 demonstrates remarkable activity against P04626 -amplified uterine serous endometrial cancer in vitro and in vivo . BACKGROUND : Uterine serous carcinomas ( USCs ) are an aggressive form of uterine cancer that may rely on P04626 /neu amplification as a driver of proliferation . The objective of this paper is to assess the sensitivity of USC cell lines with and without P04626 /neu gene amplification to afatinib , an irreversible ErbB tyrosine kinase inhibitor , and to test the efficacy of afatinib in the treatment of P04626 -amplified USC xenografts . METHODS : Eight of fifteen primary USC cell lines ( four with P04626 amplification and four without ) demonstrating similar in vitro growth rates were treated with scalar concentrations of afatinib . Effects on cell growth , signalling and cell cycle distribution were determined by flow cytometry assays . Mice harbouring xenografts of P04626 /neu-amplified USC were treated with afatinib by gavage to determine the effect on tumour growth and overall survival . RESULTS : Primary chemotherapy-resistant USC cell lines harbouring P04626 /neu gene amplification were exquisitely sensitive to afatinib exposure ( mean ± s.e.m. IC50=0.0056 ± 0.0006 μM ) and significantly more sensitive than P04626 /neu-non-amplified USC cell lines ( mean ± s.e.m. IC50=0.563 ± 0.092 μM , P < 0.0001 ) . DB08916 exposure resulted in abrogation of cell survival , inhibition of P04626 /neu autophosphorylation and S6 transcription factor phosphorylation in P04626 /neu overexpressing USC and inhibited the growth of P04626 -amplified tumour xenografts improving overall survival ( P=0.0017 ) . CONCLUSIONS : DB08916 may be highly effective against P04626 /neu-amplified chemotherapy-resistant USC . The investigation of afatinib in patients harbouring P04626 /neu-amplified USC is warranted . Management of the adverse events of afatinib : a consensus of the recommendations of the Spanish expert panel . DB08916 is an irreversible ErbB family blocker tyrosine kinase inhibitor ( TKI ) , which has recently been approved for the treatment of patients with P00533 M+ non-small cell lung cancer . As observed with reversible P00533 TKIs , it can induce class-effect adverse events . Appropriate management of afatinib-related adverse events improves quality of life and clinical outcomes in these patients . Here we provide practical recommendations for the prophylaxis and treatment of the most common of these ( e.g. , diarrhea , rash , mucositis and others ) . Signatures of positive selection in genes associated with human skin pigmentation as revealed from analyses of single nucleotide polymorphisms . Phenotypic variation between human populations in skin pigmentation correlates with latitude at the continental level . A large number of hypotheses involving genetic adaptation have been proposed to explain human variation in skin colour , but only limited genetic evidence for positive selection has been presented . To shed light on the evolutionary genetic history of human variation in skin colour we inspected 118 genes associated with skin pigmentation in the Perlegen dataset , studying single nucleotide polymorphisms ( SNPs ) , and analyzed 55 genes in detail . We identified eight genes that are associated with the melanin pathway ( Q9UMX9 , Q04671 , P17643 , P40126 , P21583 , P00533 , P14416 and Q03181 ) and presented significant differences in genetic variation between Europeans , Africans and Asians . In six of these genes we detected , by means of the EHH test , variability patterns that are compatible with the hypothesis of local positive selection in Europeans ( Q04671 , P17643 and P21583 ) and in Asians ( Q04671 , P40126 , P21583 , P00533 and P14416 ) , whereas signals were scarce in Africans ( P40126 , P00533 and P14416 ) . Furthermore , a statistically significant correlation between genotypic variation in four pigmentation candidate genes and phenotypic variation of skin colour in 51 worldwide human populations was revealed . Overall , our data also suggest that light skin colour is the derived state and is of independent origin in Europeans and Asians , whereas dark skin color seems of unique origin , reflecting the ancestral state in humans . P00533 inhibition in lung cancer : the evolving role of individualized therapy . Non-small cell lung cancer ( NSCLC ) is the major cause of cancer-related deaths in the USA and worldwide . Most patients present with advanced disease , and treatment options for these patients are generally limited to platinum-based chemotherapy and a few targeted therapies . Targeted agents currently in use for NSCLC inhibit oncogenic receptor tyrosine kinase pathways , such as the epidermal growth factor receptor ( P00533 ) pathway . While current P00533 -targeted agents , including erlotinib and gefitinib , may result in dramatic responses , they demonstrate efficacy in only a fraction of patients , and resistance to these agents frequently develops . In order to select patients most likely to benefit from blockade of P00533 pathways , investigators have focused on identifying molecular correlates of response to anti- P00533 therapy . New strategies to minimize the risk of resistance to P00533 inhibition have been employed in the development of next-generation P00533 tyrosine kinase inhibitors , such as PF00299804 and DB08916 ; these include irreversibility of target binding , inhibition of multiple P00533 family receptors , and/or simultaneous inhibition of P00533 and other oncogenic pathways . P00533 inhibitors in non-small cell lung cancer ( NSCLC ) : the emerging role of the dual irreversible P00533 / P04626 inhibitor DB08916 . Non-small cell lung cancer ( NSCLC ) is one of the most lethal types of cancer and is associated with significant mortality and morbidity worldwide . Despite improvements in conventional treatment for NSCLC , survival remains poor and improvements in patient outcome are warranted . Over recent years , basic scientific research has dramatically increased our knowledge of the pathogenesis of lung cancer and allowed us to uncover and understand the cellular pathways involved in this process . This has led to the development of therapies to selectively target these pathways . Among these , the epidermal growth factor receptor ( P00533 ) tyrosine kinase family and related downstream pathways play a critical role in cancer development and over recent years have become a validated target in NSCLC . The development of monoclonal antibodies and first-generation tyrosine kinase inhibitors ( TKIs ) targeted towards P00533 has had a considerable impact on patient outcomes . However , despite dramatic and sustained responses and the discovery of specific patient subgroups that may derive clinical benefit , resistance to first-generation P00533 TKIs inevitably develops . A new generation of agents have been developed to provide superior potency of target inhibition and further individualize the treatment of NSCLC . This article reviews P00533 -targeted therapies currently available for use and undergoing clinical development for the treatment of NSCLC , specifically focusing on next generation agents including DB08916 , an irreversible dual inhibitor of P00533 and P04626 kinases . A comprehensive review of the preclinical efficacy profile of the ErbB family blocker afatinib in cancer . DB08916 ( also known as DB08916 ) has recently been approved in several countries for the treatment of a distinct type of epidermal growth factor receptor ( P00533 ) -mutated non-small cell lung cancer . This manuscript comprehensively reviews the preclinical data on afatinib , an irreversible inhibitor of the tyrosine kinase activity of members of the epidermal growth factor receptor family ( ErbB ) including P00533 , P04626 and ErbB4 . DB08916 covalently binds to cysteine 797 of the P00533 and the corresponding cysteines 805 and 803 in P04626 and ErbB4 , respectively . Such covalent binding irreversibly inhibits the tyrosine kinase activity of these receptors , resulting in reduced auto- and transphosphorylation within the ErbB dimers and inhibition of important steps in the signal transduction of all ErbB receptor family members . DB08916 inhibits cellular growth and induces apoptosis in a wide range of cells representative for non-small cell lung cancer , breast cancer , pancreatic cancer , colorectal cancer , head and neck squamous cell cancer and several other cancer types exhibiting abnormalities of the ErbB network . This translates into tumour shrinkage in a variety of in vivo rodent models of such cancers . DB08916 retains inhibitory effects on signal transduction and in vitro and in vivo cancer cell growth in tumours resistant to reversible P00533 inhibitors , such as those exhibiting the T790M mutations . Several combination treatments have been explored to prevent and/or overcome development of resistance to afatinib , the most promising being those with P00533 - or P04626 -targeted antibodies , other tyrosine kinase inhibitors or inhibitors of downstream signalling molecules . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . P01308 secretory defects and impaired islet architecture in pancreatic beta-cell-specific P40763 knockout mice . Normal islet formation and function depends on the action of various growth factors operating in pre- and postnatal development ; however , the specific physiological function of each factor is largely unknown . Loss-of-function analyses in mice have provided little information so far , perhaps due to functional redundancies of the growth factors acting on the pancreas . The present study focuses on the role of the transcription factor P40763 in insulin-producing cells . P40763 is one of the potential downstream mediators for multiple growth factors acting on the pancreatic beta-cells , including betacellulin , hepatocyte growth factor , growth hormone , and heparin-binding P01133 -like growth factor . To elucidate its role in the beta-cells , the P40763 gene was disrupted in insulin-producing cells in mice ( P40763 -insKO ) , using a cre-mediated gene recombination approach . Unexpectedly , P40763 -insKO mice exhibited an increase in appetite and obesity at 8 weeks of age or older . The mice showed partial leptin resistance , suggesting that expression of the RIP ( rat insulin promoter ) -cre transgene in hypothalamus partially inhibited the appetite-regulating system . Intraperitoneal glucose tolerance tests , performed in non-obese 5-week-old mice , showed that the P40763 -insKO mice were glucose intolerant . Islet perifusion experiments further revealed a deficiency in early-phase insulin secretion . Whereas islet insulin content or islet mass was not affected , expression levels of P11168 , Q09428 , and P15692 were significantly reduced in P40763 -insKO islets . Interestingly , P40763 -insKO mice displayed impaired islet morphology : alpha-cells were frequently seen in central regions of islets . Our present observations demonstrate a unique role of P40763 in maintaining glucose-mediated early-phase insulin secretion and normal islet morphology . DB08916 , erlotinib and gefitinib in the first-line therapy of P00533 mutation-positive lung adenocarcinoma : a review . Non-small cell lung cancer ( NSCLC ) consists of several histomorphologically defined phenotypes that display an enormous genetic variability . In recent years , epidermal growth factor receptor ( P00533 ) mutation-positive lung adenocarcinoma has emerged as a unique subset of NSCLC in terms of etiopathogenesis and tumor biology . Since the introduction of the reversible P00533 tyrosine kinase inhibitors ( TKIs ) erlotinib and gefitinib , patients with metastatic P00533 mutation-positive lung cancer can be offered a therapeutic alternative that has proven its superiority over standard platinum-based chemotherapy . However , primary or acquired resistance limits the therapeutic success of these targeted agents . Irreversible inhibitors targeting all ErbB family receptor tyrosine kinases , such as afatinib and dacomitinib , have been developed to confer sustained disease control in ErbB-dependent cancers . The large LUX-Lung 3 phase III trial recently reported afatinib to be clearly superior over the most effective platinum doublet in patients with P00533 mutation-positive lung cancer . To fully exploit the clinical activity of afatinib , proactive management of its gastrointestinal and dermatologic toxicities is advised . [ Adverse events of afatinib as first-line treatment for five cases of advanced lung adenocarcinoma and review of literature ] . BACKGROUND AND OBJECTIVE : DB08916 is an irreversible ErbB-family blocker with a clinical activity in non-small cell lung cancer with epidermal growth factor receptor ( P00533 ) mutations . The aim of this study is to assess the safety of afatinib in patients with advanced lung adenocarcinoma . METHODS : Patients with lung adenocarcinoma ( stage IIIb or IV ) with P00533 mutations were first-line treated with an oral administration of afatinib ( 40 mg/d ) until disease progression . Adverse events , effects , and survival condition were observed . RESULTS : The most common adverse events were diarrhea ( n=5 , 100 % ) , skin rash ( n=4 , 80 % ) , and mucositis/stomatitis ( n=4 , 80 % ) . Moderate toxicities not exceeding grade 3 were observed . Relatively , the most serious adverse reaction was mucositis/stomatitis . Mild diarrhea occurred in all patients . Three patients experienced temporary drug withdrawal and dose reduction because of adverse reaction . Among the four patients who were evaluated , partial response was observed in two patients ( 50 % ) , one with stable disease ( 25 % ) and one with progressive disease ( 25 % ) . Median progression-free survival was 9.7 months , whereas median overall survival was 18.4 months . CONCLUSIONS : DB08916 was approved as first-line treatment for patients with advanced lung adenocarcinoma . The most common adverse events were diarrhea and skin rash . However , mucositis/stomatitis related to afatinib should also be considered . Considering the small number of cases , the conclusion requires more trials for confirmation . DB08916 and lung cancer . P00533 tyrosine kinase inhibitors ( P00533 -TKI ) have an established role in the treatment of non-small-cell lung cancer ( NSCLC ) . First-generation reversible DB00171 -competitive P00533 -TKIs are approved for the initial treatment of patients with P00533 mutation-positive advanced NSCLC . DB08916 is an irreversible second-generation P00533 -TKI with potent preclinical activity against P00533 ( wild type and mutant ) , P04626 , Q15303 and P00533 -mutant NSCLC with acquired resistance to reversible P00533 -TKI . LUX-Lung 3 trial demonstrated superiority of afatinib to cisplatin and pemetrexed in the frontline treatment of treatment-naïve patients with advanced adenocarcinoma of the lung and P00533 mutation . Based on these results , afatinib was recently approved for the first-line treatment of NSCLC patients with P00533 mutation . This article summarizes current status of preclinical and clinical development of afatinib in NSCLC . Oral epidermal growth factor receptor tyrosine kinase inhibitors for the treatment of non-small cell lung cancer : comparative pharmacokinetics and drug-drug interactions . The development of orally active small molecule inhibitors of the epidermal growth factor receptor ( P00533 ) has led to new treatment options for non-small cell lung cancer ( NSCLC ) . Patients with activating mutations of the P00533 gene show sensitivity to , and clinical benefit from , treatment with P00533 tyrosine kinase inhibitors ( P00533 -TKls ) . First generation reversible DB00171 -competitive P00533 -TKls , gefitinib and erlotinib , are effective as first , second-line or maintenance therapy . Despite initial benefit , most patients develop resistance within a year , 50-60 % of cases being related to the appearance of a T790M gatekeeper mutation . Newer , irreversible P00533 -TKls - afatinib and dacomitinib - covalently bind to and inhibit multiple receptors in the ErbB family ( P00533 , P04626 and Q15303 ) . These agents have been mainly evaluated for first-line treatment but also in the setting of acquired resistance to first-generation P00533 -TKls . DB08916 is the first ErbB family blocker approved for patients with NSCLC with activating P00533 mutations ; dacomitinib is in late stage clinical development . Mutant-selective P00533 inhibitors ( AZD9291 , CO-1686 , HM61713 ) that specifically target the T790M resistance mutation are in early development . The P00533 -TKIs differ in their spectrum of target kinases , reversibility of binding to P00533 receptor , pharmacokinetics and potential for drug-drug interactions , as discussed in this review . For the clinician , these differences are relevant in the setting of polymedicated patients with NSCLC , as well as from the perspective of innovative anticancer drug combination strategies . Extracellular signal-regulated kinase/mitogen-activated protein kinases block internalization of delta-opioid receptors . Translocation of G protein-coupled receptors ( GPCRs ) from the cell membrane to cytosol depends on the kind of ligand activating the receptor . This principle is clearly demonstrated for opioid receptors , because diverse opiate agonists rapidly induce receptor internalization , whereas morphine almost fails . We report here the impact of mitogen-activated protein ( Q96HU1 ) kinase isoforms extracellular signal-regulated kinase ( P29323 )1/2 on the internalization of delta-opioid receptors ( DORs ) expressed in human embryonic kidney ( P29320 )293 cells . Receptor activation by etorphine turned out to transiently phosphorylate P29323 / Q96HU1 kinases and bring about Q8IXH6 internalization within 20 min . In contrast , prolonged exposure of HEK293 cells to morphine excited persistent phosphorylation of P29323 / Q96HU1 kinases , and those cells failed to internalize the opioid receptor . When P29323 / Q96HU1 kinase phosphorylation was blocked by 2'-Amino-3'-methoxyflavone ( PD98059 ) , morphine gained the ability to strongly induce Q8IXH6 endocytosis . The importance of activated Q96HU1 kinases for Q8IXH6 internalization is further demonstrated by glutamate and paclitaxel because these substances induce phosphorylation of P27361 /2 and concomitantly prevent Q8IXH6 sequestration by etorphine . In addition , receptor internalization by morphine was facilitated by inhibition of protein kinase C and opioid-mediated transactivation of epidermal growth factor receptor ( P00533 ) , both activating P29323 / Q96HU1 kinases by opioids . The mechanism affording Q8IXH6 internalization by PD98059 may relate to arrestin , which uncouples GPCRs and thus triggers receptor internalization . Arrestin considerably translocates toward the cell membrane upon Q8IXH6 activation by morphine in presence of the Q96HU1 kinase blocker , but it fails in the absence of PD98059 . We conclude that P29323 / Q96HU1 kinase activity prevents opioid receptor desensitization and sequestration by blocking arrestin 2 interaction with activated DORs . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . DB08916 : first global approval . DB08916 , an irreversible inhibitor of the ErbB family of tyrosine kinases , is under development with Boehringer Ingelheim for the once-daily , oral treatment of cancer . DB08916 downregulates ErbB signalling by covalently binding to epidermal growth factor receptor ( P00533 ) , human epidermal growth factor receptor ( HER ) 2 and Q15303 , irreversibly inhibiting tyrosine kinase autophosphorylation . It also inhibits transphosphorylation of P21860 . Oral afatinib ( Gilotrif™ ) has been approved in the US for the first-line treatment of patients with metastatic non-small-cell lung cancer ( NSCLC ) who have tumours with P00533 exon 19 deletions or exon 21 ( L858R ) substitution mutations as detected by a US FDA-approved test . DB08916 has also been approved in Taiwan for the first-line treatment of patients with P00533 mutation-positive NSCLC . In addition , the European Medicines Agency 's Committee for Medicinal Products for Human Use has recommended the approval of afatinib ( Giotrif® ) for the treatment of patients with locally advanced or metastatic NSCLC with activating P00533 mutations who are P00533 tyrosine kinase inhibitor naïve . DB08916 is also under regulatory review in Canada , Japan and other Asian countries . This article summarizes the milestones in the development of afatinib , leading to this first approval in patients with metastatic NSCLC . Current and emerging targeted therapies for metastatic breast cancer . The success of endocrine therapies for hormone receptor-positive breast cancer and trastuzumab and lapatinib for targeting human epidermal growth factor receptor 2 ( P04626 ) -positive tumors has paved the way for the clinical development of several other metastatic breast cancer ( MBC ) -targeted therapies . Although the benefit of the anti- P15692 ( vascular endothelial growth factor ) monoclonal antibody bevacizumab in the MBC setting has become a topic of debate , clinical trial results are accumulating , and phase 3 evaluations are ongoing for newer P04626 -targeted agents ( pertuzumab and trastuzumab-maytansine immunoconjugate ) and P15692 -targeted agents ( aflibercept ) , as well as dual , epidermal growth factor receptor/ P04626 -targeted agents ( afatinib [ DB08916 ] and neratinib ) , multitargeted tyrosine kinase inhibitors ( sunitinib and pazopanib ) , and mammalian target of rapamycin ( everolimus ) and poly ( ADP-ribose ) polymerase 1 inhibitors ( iniparib , olaparib ) . These agents as well as other novel classes of anticancer agents are being tested in clinical trials with the potential of addressing unmet therapeutic needs in the MBC patient population . 20-HETE in neovascularization . Cytochrome P450 4A/F ( CYP4A/F ) converts arachidonic acid ( AA ) to 20-HETE by ω-hydroxylation . The contribution of 20-HETE to the regulation of myogenic response , blood pressure , and mitogenic actions has been well summarized . This review focuses on the emerging role of 20-HETE in physiological and pathological vascularization . 20-HETE has been shown to regulate vascular smooth muscle cells ( VSMC ) and endothelial cells ( EC ) by affecting their proliferation , migration , survival , and tube formation . Furthermore , the proliferation , migration , secretion of proangiogenic molecules ( such as HIF-1α , P15692 , SDF-1α ) , and tube formation of endothelial progenitor cells ( EPC ) are stimulated by 20-HETE . These effects are mediated through c-Src- and P00533 -mediated downstream signaling pathways , including MAPK and PI3K/Akt pathways , P29474 uncoupling , and NOX/ROS system activation . Therefore , the CYP4A/F-20-HETE system may be a therapeutic target for the treatment of abnormal angiogenic diseases . 5- Q13049 receptor induces P29323 phosphorylation and proliferation through ADAM-17 tumor necrosis factor-alpha-converting enzyme ( P78536 ) activation and heparin-bound epidermal growth factor-like growth factor ( HB- P01133 ) shedding in mesangial cells . In this study , we present multiple lines of evidence to support a critical role for heparin-bound P01133 ( epidermal growth factor ) -like growth factor ( HB- P01133 ) and tumor necrosis factor-alpha-converting enzyme ( P78536 ) ( P78536 ) in the transactivation of P01133 receptor ( P00533 ) , P29323 phosphorylation , and cellular proliferation induced by the 5-HT(2A) receptor in renal mesangial cells . 5-hydroxy-tryptamine ( 5-HT ) resulted in rapid activation of P78536 , HB- P01133 shedding , P00533 activation , P29323 phosphorylation , and longer term increases in DNA content in mesangial cells . P29323 phosphorylation was attenuated by 1 ) neutralizing P00533 antibodies and the P00533 kinase inhibitor , AG1478 , 2 ) neutralizing HB- P01133 , but not amphiregulin , antibodies , heparin , or CM197 , and 3 ) pharmacological inhibitors of matrix-degrading metalloproteinases or P78536 small interfering RNA . Exogenously administered HB- P01133 stimulated P29323 phosphorylation . Additionally , P78536 was co-immunoprecipitated with HB- P01133 . Small interfering RNA against P78536 also blocked 5-HT-induced increases in P29323 phosphorylation , HB- P01133 shedding , and DNA content . In aggregate , this work supports a pathway map that can be depicted as follows : 5-HT --> 5-HT(2A) receptor --> P78536 --> HB- P01133 shedding --> P00533 --> P29323 --> increased DNA content . To our knowledge , this is the first time that P78536 has been implicated in 5-HT-induced P00533 transactivation or in proliferation induced by a G protein-coupled receptor in native cells in culture . P01308 -like growth factor-1 receptor and ligand targeting in head and neck squamous cell carcinoma . DB01277 receptor ( IGF-1R ) signaling is associated with increased tumorigenesis of epithelial cancers . In light of recent epidemiological studies correlating high circulating levels of DB01277 with increased risk of second primary tumors ( SPTs ) of the head and neck , we examined IGF system and epidermal growth factor receptor ( P00533 ) expression in human head and neck squamous cell carcinoma ( HNSCC ) matched pairs and a cross-section of HNSCC cell lines . Employing the latter , we demonstrated that DB01277 stimulated S-phase transition in a PI 3-K/Akt and Erk-dependent manner in 5 of 8 cell lines , with Erk activation being dependent upon P00533 kinase activity . DB01277 stimulated thymidine incorporation was inhibited by treatment with P17936 , the IGF-1R tyrosine kinase inhibitor DB00238 -AEW541 , or the P00533 tyrosine kinase inhibitor AG1478 . Combining P17936 with DB00238 -AEW541 or AG1478 abrogated DB01277 responses at 10-fold lower doses than either compound alone . These results demonstrate the potential for co-targeting the IGF system and P00533 in HNSCC . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Diarrhea associated with afatinib : an oral ErbB family blocker . Gastrointestinal ( GI ) adverse events ( AEs ) are frequently observed in patients receiving P01133 receptor ( P00533 ; also known as P00533 or ErbB1 ) tyrosine kinase inhibitor therapy . GI AEs are among the most common and most impactful on a patient 's quality of life . Severe diarrhea can result in fluid and electrolyte losses , leading to dehydration , electrolyte imbalances and renal insufficiency . DB08916 is an irreversible , oral , ErbB family blocker , inhibiting P00533 ( ErbB1 ) , P04626 ( ErbB2 ) and ErbB4 receptor kinases . It also inhibits transphosphorylation of ErbB3 . Similar to reversible tyrosine kinase inhibitors of P00533 , GI AEs - in particular , diarrhea - have frequently been observed in afatinib-treated patients . This article summarizes current data on afatinib-associated diarrhea and provides strategies for its management . Patient education , early identification , timely management and ongoing assessment will help to prevent aggravation , afatinib dose reductions or therapy discontinuation , encouraging patient compliance and allowing patients to obtain the maximum therapeutic benefit from this agent . [ A new perspective in the treatment of non-small-cell lung cancer (NSCLC). Role of afatinib : An oral and irreversible ErbB family blocker ] . Tyrosine kinase inhibitors ( TKI ) that block epidermal growth factor receptor ( P00533 ) pathway have demonstrated a clinical benefit for patients with non-small-cell lung cancer ( NSCLC ) harboring P00533 mutations . The currently available TKI ( gefitinib and erlotinib ) are P00533 reversible inhibitors . DB08916 is an oral , irreversible ErbB family blocker that covalently binds and blocks signaling from P00533 ( ErbB1 ) , P04626 ( ErbB2 ) and ErbB4 . The compound inhibits also the transphosphorylation of ErbB3 . With this mode of action , afatinib is thought to have a mechanistic advantage over P00533 blockade alone , in that it provides a sustained , covalent inhibition of ErbB homo- and hetero-dimers . In the pivotal LUX-Lung 3 study , afatinib demonstrated a prolonged progression free survival over standard pemetrexed plus cisplatin chemotherapy ( 11.1 versus 6.9 months ; HR = 0.58 , 95 % CI : 0.43-0.78 ; P = 0.001 ) in P00533 mutation positive NSCLC patients . The compound has recently been granted a marketing authorization ( MA ) for the treatment of patients with locally advanced or metastatic NSCLC with activating P00533 mutation(s) and P00533 TKI-naive . In this paper are summarized the efficacy and safety data in this indication . DB08916 for Erlotinib Refractory Brain Metastases in a Patient with P00533 -Mutant Non-Small-Cell Lung Cancer : Can High-Affinity TKI Substitute for High-Dose TKI ? No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . Convergent and divergent cellular responses by ErbB4 isoforms in mammary epithelial cells . Associations of ErbB4 ( Q15303 / Q15303 ) , the fourth member of the P00533 family , with cancer are variable , possibly as a result of structural diversity of this receptor . There are multiple structural isoforms of Q15303 arising by alternative mRNA splicing , and a subset undergo proteolysis that releases membrane-anchored and soluble isoforms that associate with transcription factors and coregulators to modulate transcription . To compare the differential and common signaling activities of full-length ( FL ) and soluble intracellular isoforms of Q15303 , four JM-a isoforms ( FL and soluble intracellular domain ( ICD ) CYT-1 and CYT-2 ) were expressed in isogenic MCF10A cells and their biologic activities were analyzed . Both FL and ICD CYT-2 promoted cell proliferation and invasion , and CYT-1 suppressed cell growth . Transcriptional profiling revealed several new and underexplored Q15303 -regulated transcripts , including : proteases/protease inhibitors ( P08254 and P07093 ) , the YAP/Hippo pathway ( P29279 , O00622 , and P09486 ) , the mevalonate/cholesterol pathway ( P04035 , Q01581 , P01130 , and Q9UBM7 ) , and cytokines ( P10145 , P78556 , and P09341 ) . Many of these transcripts were subsequently validated in a luminal breast cancer cell line that normally expresses Q15303 . Furthermore , ChIP-seq experiments identified O75689 , P02649 , P09486 , P16949 , and Q05195 as novel molecular targets of Q15303 . These findings clarify the diverse biologic activities of Q15303 isoforms , and reveal new and divergent functions . IMPLICATIONS : ErbB4 as a regulator of Hippo and mevalonate pathways provides new insight into milk production and anabolic processes in normal mammary epithelia and cancer . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . Acquired resistance to epidermal growth factor receptor kinase inhibitors associated with a novel T854A mutation in a patient with P00533 -mutant lung adenocarcinoma . PURPOSE : Somatic mutations in the tyrosine kinase domain of the epidermal growth factor receptor ( P00533 ) gene are associated with sensitivity of lung adenocarcinomas to the P00533 tyrosine kinase inhibitors , gefitinib and erlotinib . Acquired drug resistance is frequently associated with a secondary somatic mutation that leads to the substitution of methionine for threonine at position 790 ( T790M ) . We aimed to identify additional second-site alterations associated with acquired resistance . EXPERIMENTAL DESIGN : Tumor samples were obtained from 48 patients with acquired resistance . Tumor cell DNA was analyzed for P00533 kinase domain mutations . Molecular analyses were then done to characterize the biological properties of a novel mutant P00533 allele . RESULTS : A previously unreported mutation in exon 21 of P00533 , which leads to substitution of alanine for threonine at position 854 ( T854A ) , was identified in one patient with a drug-sensitive P00533 L858R-mutant lung adenocarcinoma after long-term treatment with tyrosine kinase inhibitors . The T854A mutation was not detected in a pretreatment tumor sample . The crystal structure analyses of P00533 suggest that the T854 side chain is within contact distance of gefitinib and erlotinib . Surrogate kinase assays show that the P00533 T854A mutation abrogates the inhibition of tyrosine phosphorylation by erlotinib . Such resistance seems to be overcome by a new irreversible dual P00533 / P04626 inhibitor , DB08916 . CONCLUSIONS : The T854A mutation is the second reported second-site acquired resistance mutation that is within contact distance of gefitinib and erlotinib . These data suggest that acquired resistance to DB00171 -mimetic P00533 kinase inhibitors may often be associated with amino acid substitutions that alter drug contact residues in the P00533 DB00171 -binding pocket . Boca-dependent maturation of beta-propeller/ P01133 modules in low-density lipoprotein receptor proteins . The extracellular portions of cell surface receptor proteins are often comprised of independently folding protein domains . As they are translated into the endoplasmic reticulum ( ER ) , some of these domains require protein chaperones to assist in their folding . Members of the low-density lipoprotein receptor ( P01130 ) family require the chaperone called Boca in Drosophila or its ortholog , Mesoderm development , in the mouse . All LDLRs have at least one six-bladed beta-propeller domain , which is immediately followed by an epidermal growth factor ( P01133 ) repeat . We show here that Boca is specifically required for the maturation of these beta-propeller/ P01133 modules through the secretory pathway , but is not required for other P01130 domains . Protein interaction data suggest that as LDLRs are translated into the ER , Boca binds to the beta-propeller . Subsequently , once the P01133 repeat is translated , the beta-propeller/ P01133 module achieves a more mature state that has lower affinity for Boca . We also show that Boca-dependent beta-propeller/ P01133 modules are found not only throughout the P01130 family but also in the precursor to the mammalian P01133 ligand . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . DB08916 in NSCLC harbouring P00533 mutations . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Dermatologic adverse events associated with afatinib : an oral ErbB family blocker . Dermatologic adverse events ( AEs ) are frequently observed in patients receiving P01133 receptor ( P00533 ; also known as ErbB1 ) tyrosine kinase inhibitor therapy . The impact of these AEs goes beyond cosmesis to the discomfort from itching , pain and secondary infections , all of which may significantly impact on patient well-being , adherence and clinical outcomes . DB08916 is a potent , irreversible , oral , ErbB family blocker , inhibiting P00533 ( ErbB1 ) , P04626 ( ErbB2 ) and ErbB4 receptor kinases . It also inhibits transphosphorylation of ErbB3 . Similar to P00533 inhibitors , dermatologic AEs have been frequently observed in patients treated with afatinib . Papulopustular ( acneiform ) rash , pruritus , xerosis , paronychia and alopecia will require patient education and proactive treatment interventions . This article summarizes current data on the dermatologic AEs associated with afatinib treatment across the clinical trial program , and provides strategies for their effective management . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . DB08916 in the treatment of breast cancer . INTRODUCTION : DB00072 has dramatically improved outcomes for those diagnosed with human EGFR2 ( P04626 ) -positive breast cancer . Resistance to trastuzumab , however , is an ongoing problem that has led to the development of a number of new P04626 -targeted therapies . DB08916 is a novel , orally bioavailable irreversible pan-HER inhibitor that has been evaluated in multiple tumor types . It has also shown promise in NSCLC where it has earned FDA approval . Its activity in breast cancer is currently being evaluated . AREAS COVERED : This review briefly summarizes the current therapies available for P04626 -positive metastatic disease . This article also describes the data available for afatinib in breast cancer from preclinical analyses , published Phase I and II trials to ongoing and upcoming Phase II and III studies . EXPERT OPINION : While Phase I and II studies have demonstrated promising activity in P04626 -positive breast cancer , the Phase III randomized study of afatinib in trastuzumab-resistant metastatic breast cancer was halted early due to unfavorable risk-benefit analysis from the Independent Data Monitoring Committee ( IDMC ) . The successful development of afatinib in breast cancer will thus depend on aggressively preventing and managing its associated toxicities . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . The role of irreversible HER family inhibition in the treatment of patients with non-small cell lung cancer . Small-molecule tyrosine kinase inhibitors ( TKIs ) of the human epidermal growth factor receptor ( HER ) include the reversible epidermal growth factor receptor ( P00533 /HER-1 ) inhibitors gefitinib and erlotinib . P00533 TKIs have demonstrated activity in the treatment of patients with non-small cell lung cancer ( NSCLC ) harboring activating P00533 mutations ; however , multiple mechanisms of resistance limit the benefit of these drugs . Although resistance to P00533 TKIs can be intrinsic and correlated with molecular lesions such as in Kirsten rat sarcoma viral oncogene homolog ( P01116 ; generally observed in a wild-type P00533 background ) , acquired resistance to P00533 TKIs can evolve in the setting of activating P00533 mutations , such as in the case of P00533 T790M mutations . Several irreversible inhibitors that target multiple members of the HER family simultaneously are currently in clinical development for NSCLC and may have a role in the treatment of TKI-sensitive and TKI-resistant disease . These include PF00299804 , an inhibitor of P00533 /HER-1 , HER-2 , and HER-4 , and afatinib ( DB08916 ) , an inhibitor of P00533 /HER-1 , HER-2 , and HER-4 . Results of large , randomized trials of these agents may help to determine their potential for the treatment of NSCLC . Glioma cell activation by Alzheimer 's peptide Abeta1-42 , alpha1-antichymotrypsin , and their mixture . We compared the effects ofAlzheimer 's peptide ( Abeta1-42 ) , a,-antichymotrypsin ( ACT ) and an ACT/Abeta1-42 mixture on human glioma DK-MG cells . The solution of Abeta ( 5 microM ) formed by 2-h incubation at room temperature induced tumour necrosis factor-alpha ( P01375 ) and interleukin ( IL ) -6 levels by 55 and 45 % , respectively , and increased gelatinase B activity by 67 % , while exposure of cells to the ACT/Abeta1-42 mixture ( 1:10 molar ratio ACT : Abeta1-42 ) under the same experimental conditions showed no effect on P05231 levels or gelatinase B activity , but strongly induced P01375 ( by 190 % ) , compared to the controls . Stimulation of the cells with Abeta1-42 alone , but not with ACT , increased by about 20 % low-density lipoprotein ( LDL ) uptake and mRNA levels for P01130 and P04035 , while the ACT/Abeta1-42 mixture significantly increased LDL uptake ( by 50 % ) , up-regulated mRNA levels for P01130 and P04035 by 48 and 63 % , respectively , and increased lipid accumulation by about 20-fold . These data suggest a possible new role for Abeta in Alzheimer 's disease through its interaction with the inflammatory reactant , ACT . P01308 -like growth factor-1 receptor as a novel prognostic marker and its implication as a cotarget in the treatment of human adenocarcinoma of the esophagus . P01308 -like growth factor-1 receptor ( IGF-1R ) and human epidermal growth factor receptor-2 ( P04626 ) receptor expression has been found to be a key regulator of tumorigenesis . The purpose of our study was to establish the prognostic significance of IGF-1R in esophageal cancer and to determine the effect of IGF-1R and P04626 targeting with alpha-IR3 and Herceptin antibodies on the proliferation of esophageal cancer cells in vitro . IGF-1R expression and clinicopathological correlations were analyzed with a tissue microarray containing 234 esophageal cancer specimens ( 133 adenocarcinomas and 101 squamous cell carcinomas ) . Proliferation changes associated with Herceptin and alpha-IR3 blockage were evaluated with the unique human esophageal cancer cell lines Pt1590 and LN1590 . IGF-1R and P04626 expression levels , activation and phosphorylation status of downstream signaling proteins involved in the activation pathways were analyzed by Western blotting . IGF-1R overexpression was detected in 121 ( 52 % ) of the 234 esophageal tumors examined . In the subgroup of 87 P04626 -positive tumors , 93.1 % showed concordant overexpression for IGF-1R . IGF-1R was identified as a variable associated with reduced overall survival for adenocarcinoma ( p = 0.05 ) , but not for squamous cell carcinoma . The combination of Herceptin and alpha-IR3 was more effective in inhibiting in vitro proliferation than treatment with either agent alone ( p < 0.01 ) . This was associated with a decrease in P04626 and IGF-1R protein levels and suppression of Akt- and Q96HU1 kinase phosphorylation . IGF-1R expression can be used as a novel prognostic marker for adenocarcinomas of the esophagus . Cotreatment with IGF-1R and P04626 antibodies might become a valuable and effective treatment option in esophageal adenocarcinoma . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . [ Cellular adhesion signal transduction network of tumor necrosis factor-alpha induced hepatocellular carcinoma cells ] . OBJECTIVE : To systemically explore the cellular adhesion signal transduction network of tumor necrosis factor-alpha ( P01375 -α ) -induced hepatocellular carcinoma cells with bioinformatics tools . METHODS : Published microarray dataset of P01375 -α-induced HepG2 , human transcription factor database HTRI and human protein-protein interaction database HPRD were used to construct and analyze the signal transduction network . RESULTS : In the signal transduction network , MYC and SP1 were the key nodes of signaling transduction . Several genes from the network were closely related with cellular adhesion. P00533 ( P00533 ) is a possible key gene of effectively regulating cellular adhesion during the induction of P01375 -α . CONCLUSION : P00533 is a possible key gene for P01375 -α-induced metastasis of hepatocellular carcinoma . A phase I dose escalation study of DB08916 , an irreversible dual inhibitor of epidermal growth factor receptor 1 ( P00533 ) and 2 ( P04626 ) tyrosine kinase in a 2-week on , 2-week off schedule in patients with advanced solid tumours . To assess tolerability , pharmacokinetics ( PK ) , pharmacodynamics ( PD ) and clinical activity of the dual epidermal growth factor receptor ( P00533 ) 1 and 2 ( P04626 ) tyrosine kinase inhibitor DB08916 . An escalating schedule of once-daily ( OD ) DB08916 for 14 days followed by 14 days off medication was explored . Thirty-eight patients were enrolled . Dose levels were 10 , 20 , 30 , 45 , 70 , 85 , and 100 mg . At 100 mg dose-limiting toxicity ( DLT ) ( common toxicity criteria grade 3 skin rash and grade 3 diarrhoea despite treatment with loperamide ) occurred in two patients . In the next-lower dose of 70 mg , DLT ( grade 3 fatigue and ALAT elevation ) occurred in one of six patients . An intermediate dose level of 85 mg was studied . Here DLT occurred in two patients ( grade 3 diarrhoea despite treatment and grade 2 diarrhoea lasting more than 7 days despite treatment ) . An additional 12 patients were treated at 70 mg . DB08916 PK after single and multiple doses revealed moderately fast absorption , and no deviation from dose proportionality . Pharmacodynamics analysis in skin biopsies did not show significant changes in P00533 -associated biomarkers . However , a significant inhibitory effect on the proliferation index of epidermal keratinocytes was observed . No partial or complete responses were observed , stable disease lasting more than four cycles was seen in seven patients . The recommended dose for studies with DB08916 for 14 days followed by 14 days off medication is 70 mg OD . Next generation tyrosine kinase inhibitor ( TKI ) : afatinib . DB08916 is a recently introduced new tyrosine kinase inhibitor , approved by the USFDA on July 12 , 2013 . DB08916 is marketed under the trade name Gilotrif and developed by Boehringer Ingelheim GmbH . It is indicated for the first-line treatment of patients with metastatic non-small cell lung cancer ( NSCLC ) carrying P00533 exon 19 deletions or exon 21 ( L858R ) mutations . DB08916 is a covalent , irreversible inhibitor of epidermal growth factor receptor ( P00533 ) , human epidermal growth factor receptor 2 ( P04626 ) and Q15303 . Chemically afatinib is a 4-anilinoquinazoline derivative , having an acrylamide warhead . Gilotrif is the formulation of DB08916 di-meleate salt . Presently , afatinib has been approved in the USA , the European Union , Taiwan and Mexico . In this review , we have summarized the chemical characterization of afatinib , its synthesis , patent status , marketed formulation , available crystalline form and current clinical trials . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . Development of [18F]afatinib as new TKI-PET tracer for P00533 positive tumors . INTRODUCTION : DB08916 is an irreversible ErbB family blocker that was approved for the treatment of P00533 mutated non-small cell lung cancer in 2013 . Positron emission tomography ( PET ) with fluorine-18 labeled afatinib provides a means to obtain improved understanding of afatinib tumor disposition in vivo . PET imaging with [(18)F]afatinib may also provide a method to select treatment responsive patients . The aim of this study was to label afatinib with fluorine-18 and evaluate its potential as TKI-PET tracer in tumor bearing mice . METHODS : A radiochemically novel coupling , using peptide coupling reagent BOP , was explored and optimized to synthesize [(18)F]afatinib , followed by a metabolite analysis and biodistribution studies in two clinically relevant lung cancer cell lines , xenografted in nude mice . RESULTS : A reliable [(18)F]afatinib radiosynthesis was developed and the tracer could be produced in yields of 17.0 ± 2.5 % calculated from [(18)F]F(-) and > 98 % purity . The identity of the product was confirmed by co-injection on HPLC with non-labeled afatinib . Metabolite analysis revealed a moderate rate of metabolism , with > 80 % intact tracer in plasma at 45 min p.i . Biodistribution studies revealed rapid tumor accumulation and good retention for a period of at least 2 hours , while background tissues showed rapid clearance of the tracer . CONCLUSION : We have developed a method to synthesize [(18)F]afatinib and related fluorine-18 labeled 4-anilinoquinazolines . [(18)F] DB08916 showed good stability in vivo , justifying further evaluation as a TKI-PET tracer . Positive and negative selection for tumor necrosis factor responsiveness reveals an inhibitory role for P01133 receptor in P01375 -induced antiproliferation . P01375 ( P01375 ) induces dose-dependent , but incomplete cytotoxicity in ME-180 cervical carcinoma cells resulting in a significant reduction in cell viability . In this cell line there exists a characteristic residual tumor cell population that appears to be resistant to P01375 . In order to investigate tumor cell heterogeneity and characteristics that correlate with their escape from P01375 -induced cytotoxicity , P01375 -resistant ME-180 cell variants ( ME-180R ) were isolated from a population of ME-180 cervical carcinoma cells ( ME-180 parental ) . Incubation of ME-180 parental cells with P01375 resulted in measurable changes in tumor cell DNA structural integrity and dose-dependent cytotoxicity , whereas ME-180R cell growth and DNA integrity were not effected by incubation with P01375 . Binding of 125I-labeled P01375 to a P01375 -specific cell-surface receptor was measurable and equivalent on both ME-180R and ME-180 parental cells and both cell lines predominantly expressed the p55 form of the P01375 receptor based upon flow cytometric analysis . Although both cell lines shared similar doubling times , intrinsic P01133 receptor tyrosine kinase activity in ME-180R cells was found to be > 3-fold higher than that isolated from ME-180 parental cells . These results suggest that P01375 -responsiveness may be mediated at a point subsequent to P01375 binding and may be regulated , in part , by the expression of tyrosine kinase activity . To further explore this hypothesis , A431 vulvular carcinoma cells that express resistance to P01375 were cloned and variants were isolated that escaped P01133 -induced growth inhibition. ( ABSTRACT TRUNCATED AT 250 WORDS ) LUX-Lung 3 : redundancy , toxicity or a major step forward ? DB08916 as front-line therapy for patients with metastatic P00533 -mutated lung cancer . Mutant P01133 receptor ( P00533 ) is an attractive therapeutic target in patients with metastatic non-small cell lung cancer ( NSCLC ) . A new paradigm has been defined using DB00171 -competitive P00533 tyrosine kinase inhibitors ( TKIs ) , gefitinib and erlotinib , as the most effective first-line treatment . However , clinical benefit of P00533 -TKI is only transient and limited by primary or acquired resistance . DB08916 has been developed as a highly potent , irreversible inhibitor of P00533 , P04626 and ErbB4 , as well as transphosphorylation of ErbB3 . The clinical activity of afatinib in lung cancer has been extensively studied in the LUX-Lung series of trials . LUX-Lung 3 was a pivotal randomized Phase III study that recently led to the approval of afatinib ( Gilotrif ) in several countries for treatment of patients with metastatic NSCLC harboring somatic P00533 gene mutations . Here , we review the rationale , study design including end points , results and implications of this trial for current and future P00533 genotype-directed lung cancer therapies . DB08916 circumvents multidrug resistance via dually inhibiting DB00171 binding cassette subfamily G member 2 in vitro and in vivo . Multidrug resistance ( MDR ) to chemotherapeutic drugs is a formidable barrier to the success of cancer chemotherapy . Expressions of DB00171 -binding cassette ( DB01048 ) transporters contribute to clinical MDR phenotype . In this study , we found that afatinib , a small molecule tyrosine kinase inhibitor ( TKI ) targeting P00533 , HER-2 and HER-4 , reversed the chemoresistance mediated by Q9UNQ0 in vitro , but had no effect on that mediated by multidrug resistance protein P08183 and P33527 . In addition , afatinib , in combination with topotecan , significantly inhibited the growth of Q9UNQ0 - overexpressing cell xenograft tumors in vivo . Mechanistic investigations exhibited that afatinib significantly inhibited ATPase activity of Q9UNQ0 and downregulated expression level of Q9UNQ0 , which resulted in the suppression of efflux activity of Q9UNQ0 in parallel to the increase of intracellular accumulation of Q9UNQ0 substrate anticancer agents . Taken together , our findings may provide a new and useful combinational therapeutic strategy of afatinib with chemotherapeutical drug for the patients with Q9UNQ0 overexpressing cancer cells . Activation of IL-6R/ P23458 / P40763 signaling induces de novo resistance to irreversible P00533 inhibitors in non-small cell lung cancer with T790M resistance mutation . The secondary T790M mutation in epidermal growth factor receptor ( P00533 ) is the major mechanism of acquired resistance to P00533 tyrosine kinase inhibitors ( TKI ) in non-small cell lung cancer ( NSCLC ) . Although irreversible P00533 TKIs , such as afatinib or dacomitinib , have been introduced to overcome the acquired resistance , they showed a limited efficacy in NSCLC with T790M . Herein , we identified the novel de novo resistance mechanism to irreversible P00533 TKIs in H1975 and Q8NBP7 -GR cells , which are NSCLC cells with P00533 T790M . DB08916 activated interleukin-6 receptor ( IL-6R ) / P23458 / P40763 signaling via autocrine P05231 secretion in both cells . Inhibition of IL-6R/ P23458 / P40763 signaling pathway increased the sensitivity to afatinib . Cancer cells showed stronger P40763 activation and enhanced resistance to afatinib in the presence of MRC5 lung fibroblasts . Blockade of IL-6R/ P23458 significantly increased the sensitivity to afatinib through inhibition of afatinib-induced P40763 activation augmented by the interaction with fibroblasts , suggesting a critical role of paracrine IL-6R/ P23458 / P40763 loop between fibroblasts and cancer cells in the development of drug resistance . The enhancement of afatinib sensitivity by inhibition of IL-6R/ P23458 / P40763 signaling was confirmed in in vivo Q8NBP7 -GR xenograft model . Similar to afatinib , de novo resistance to dacomitinib in H1975 and Q8NBP7 -GR cells was also mediated by dacomitinib-induced P23458 / P40763 activation . Taken together , these findings suggest that IL-6R/ P23458 / P40763 signaling can be a potential therapeutic target to enhance the efficacy of irreversible P00533 TKIs in patients with P00533 T790M . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility . Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor ( P14416 ) in flexible behavioral adaptation , mostly shown in reward-based learning paradigms . Recent evidence from imaging genetics indicates that also intentional cognitive flexibility , associated with lateral frontal cortex , is affected by variations in P14416 signaling . In the present functional magnetic resonance imaging ( Q9BWK5 ) study , we tested the effects of a direct pharmacological manipulation of P14416 stimulation on intentional flexibility in a task-switching context , requiring switches between cognitive task rules and between response hands . In a double blind , counterbalanced design , participants received either a low dose of the P14416 agonist bromocriptine or a placebo in two separate sessions . DB01200 modulated the blood-oxygen-level-dependent ( BOLD ) signal during rule switching : rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo , at comparable performance levels . Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions . Hand-switching-related activity , in contrast , was reduced under bromocriptine in sensorimotor regions . Our results provide converging evidence for an involvement of P14416 signaling in fronto-striatal mechanisms underlying intentional flexibility , and indicate that the neural mechanisms underlying different types of flexibility ( cognitive vs motor ) are affected differently by increased dopaminergic stimulation . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . [ DB08916 ( DB08916 ) ] . DB08916 ( DB08916 ) is an irreversible multi-target HER receptor tyrosine kinase inhibitor developed in patients with advanced solid tumours . Several phase I studies were conducted in patients with non-small cell lung cancer ( NSCLC ) , as a single agent or in combination . In further phase II or III studies , patients were selected based on the duration of response to first generation P00533 -TKI in previous line ( supposed to have greater chance to have an activating P00533 mutation ) or based directly on the P00533 activating mutation status . Here , we report and comment the main results of these studies in lung cancer patients . This drug has been approved by the Food and Drug Administration in June 2013 for the first-line treatment of patients with metastatic NSCLC whose tumours have P00533 mutation . In Europe , it has been approved in September 2013 in the same indication . [ Hange-Shashin-to for preventing diarrhea during afatinib therapy ] . DB08916 is an epidermal growth factor receptor-tyrosine kinase inhibitor( P00533 -TKI) . In a randomized phase III study ( Lux- Lung 3 study ) employing patients harboring P00533 mutations , patients administered afatinib show a significantly longer progression free survival time(PFS)than those administeredcombination chemotherapy comprising cisplatin andpemetrexed . However , most of the patients ( 95.2 % ) treatedwith afatinib experiencedd iarrhea . In the present report , 16 patients with P00533 mutations were treatedby afatinib at our institution from May 2014 to December 2014 . Twelve patients were administered a diarrhea prevention herbal medicine , Hange-shashin-to . Seven of 12 patients ( 58 % ) had no diarrhea during the 28 days of therapy . All 4 of the patients who did not receive Hange-shashin-to experienced diarrhea above Grade 1 within 6 days of starting therapy . The rate of diarrhea differed significantly between the patients receiving and not receiving Hangeshashin- to . In conclusion , preventive administration of Hange-shashin-to may reduce the occurrence of diarrhea during afatinib treatment .

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interacts_with DB00181?

multiple_choice

Transmitter neurochemistry of the efferent neuron system innervating the labyrinth . It is likely that several mechanisms contribute to the efferent control of cochlear and vestibular function . Different effects are probably mediated by different neuronal transmitters . In spite of a number of transmitter candidates , it is still widely assumed that the entire efferent system can be globally characterized as cholinergic . We attempted to label retrogradely identified efferent neurons in the brainstem with a monoclonal antibody against choline acetyltransferase ( P28329 ) , the acetylcholine ( ACh ) synthesizing enzyme . Only a portion of the vestibular efferents could thus be shown to be cholinergic in the rat . Medial cochlear efferents , terminating under outer hair cells , may also be cholinergic since they stain intensely for acetylcholine esterase ( P22303 ) after pre-treatment with the P22303 inhibitor diisopropylfluorophosphate ( DB00677 ) . The lateral cochlear efferents terminating under inner hair cells , as well as more than half of the vestibular efferent neuron population , reacted negatively with either method designed to identify cholinergic neurons . Half of the lateral olivo-cochlear neuron population filled retrogradely with tritiated gamma-amino butyric acid [ ( 3H ] -GABA ) . These cells were similar in size and distribution to neurons staining for the GABA synthesizing enzyme glutamic acid decarboxylase ( Q99259 ) . Retrograde transport of [ 3H ] -aspartate from the inner ear to the brainstem was seen in half of the lateral olivocochlear population , as well as in part of the efferent vestibular population in group E and in the caudal pontine reticular nucleus ( P16435 ) . Since various peptides have also been located in efferent neurons , this system is chemically diversified . Several distinct mechanisms of efferent control with presumably differing functions must , therefore , exist . 5- Q9H205 - and P28335 -antagonist properties of cyamemazine : significance for its clinical anxiolytic activity . RATIONALE : DB09000 is a neuroleptic compound which possesses anxiolytic properties in humans . On the other hand , 5- Q9H205 - and P28335 -receptors have been implicated in anxiety disorders and a previous binding study has shown that cyamemazine possesses high affinity for both serotonin receptor types . OBJECTIVE : The present study was undertaken to establish whether cyamemazine antagonizes 5- Q9H205 - and/or P28335 -mediated responses , and whether it compares with reference compounds . METHODS : DB09000 was tested for its ability to antagonize : ( i ) 5- Q9H205 -dependent contraction of the isolated guinea-pig ileum and bradycardic responses in the rat and ( ii ) P28335 -dependent phospholipase C ( P98160 ) stimulation in rat brain membranes . RESULTS : In isolated guinea-pig ileum , cyamemazine potently and competitively antagonized 5-HT-dependent contractions ( pA2 = 7.52 +/- 0.08 ; n = 5 ) . In this test , cyamemazine was 5-7 times more potent ( pIC50 = 6.75 +/- 0.13 ) than tropisetron ( pIC50 = 6.02 +/- 0.04 ) . In rats , cyamemazine i.v. antagonized 5-HT-dependent bradycardic responses with ID50 % = 3.2 +/- 1.5 mg/kg ( n = 4 ) . Finally , in rat brain membranes cyamemazine antagonized P28335 -dependent P98160 stimulation with Ki = 424 nM ( mianserin exhibits a Ki = 113 nM ) . CONCLUSIONS : DB09000 behaves as an antagonist at both 5- Q9H205 - and P28335 -receptors , which compares well with reference compounds . These 5- Q9H205 - and P28335 -antagonistic actions of cyamemazine can be involved , at least in part , in its beneficial therapeutic actions in anxiety disorders . The heteromeric GABA-B receptor recognizes G-protein alpha subunit C-termini . The recently cloned GABA-B receptors are related to the metabotropic glutamate receptors ( mGlu receptors ) , the Ca2+-sensing receptor and one group of vomeronasal receptors . The GABA-B receptors likely function in a heterodimeric form , constituted of Q9UBS5 and O75899 . This novel feature in the G-protein coupled receptors ( GPCRs ) structure raises questions as to the mechanism of recognition of G-proteins by such receptors . In the present study we show that the Q9UBS5 and BR2 subunits form a functional receptor that recognizes the extreme C-termini of the G alpha i and G alpha o proteins when expressed in HEK293 cells . Indeed , heteromeric Q9UBS5 /BR2 receptors do not activate P98160 when co-expressed with G alpha q , but do so when co-expressed with the chimeric G alpha qi5 or G alpha qo5 subunits , the G alpha q subunit in which the 5 C-terminal residues are those of G alpha i or G alpha o , respectively . Interestingly , the heteromeric GABA-B receptor did not activate the chimeric G alpha qz5 subunit that contains the 5 C-terminal residues of G alpha z . Among the three residues that are distinct between G alpha qo5 and G alpha qz5 ( at position -5 , -4 and -1 ) , the amino acid residue at position -4 of G alpha o proteins is critical for specifying the coupling selectivity with the receptor and residue -5 influences the coupling efficacy . Interestingly , these findings correspond to data obtained with the Q14416 receptor , a distant relative of GABA-B proteins . This shows that the same molecular determinants of the G-protein alpha-subunits are involved in the specific recognition of both the heteromeric GABA-B receptors and the other GPCRs . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Identification of genes associated with enhanced metastasis of a large cell lung carcinoma cell line . We have compared the transcriptional profile of large cell lung tumor cell lines NIH-H460 and H460-M making use of the Affymetrix GeneChip system . H460-M is derived from NIH-H460 and displays enhanced experimental and spontaneous metastasis in nude mice . Out of the 12,600 genes investigated , 73 ( 0.6 % ) were up-regulated and 114 ( 0.9 % ) were down-regulated on the basis of the scoring criteria . We have classified the de-regulated genes according to the following categories : immune response , enzymes , modulation of transcription , signal transduction , cytoskeleton/adhesion and extracellular matrix associated proteins , cell-cycle/apoptosis , transporters and ' others ' . Among the remarkable features of this system are the up-regulation of the steady-state mRNA levels of neuroendocrine markers such as neurotensin ( P30990 ) , neuroendocrine-specific protein ( NSP ) , neural cell adhesion molecule 1 ( P13591 ) and gamma-aminobutyric acid B-type receptor ( O75899 ) in cell line H460-M . Semaphorin 3B ( Q13214 ) was dramatically down-regulated in cell line H460-M and emerged as the most interesting gene for target validation . The antimalarial drug mefloquine inhibits cardiac inward rectifier K+ channels : evidence for interference in PIP2-channel interaction . The antimalarial drug mefloquine was found to inhibit the KATP channel by an unknown mechanism . Because mefloquine is a Cationic amphiphilic drug and is known to insert into lipid bilayers , we postulate that mefloquine interferes with the interaction between PIP2 and Kir channels resulting in channel inhibition . We studied the inhibitory effects of mefloquine on Kir2.1 , Kir2.3 , Kir2.3(I213L) , and Kir6.2/SUR2A channels expressed in P29320 -293 cells , and on IK1 and Q14654 from feline cardiac myocytes . The order of mefloquine inhibition was Kir6.2/SUR2A ≈ Kir2.3 ( IC50 ≈ 2 μM ) > Kir2.1 ( IC50 > 30 μM ) . Similar results were obtained in cardiac myocytes . The Kir2.3(I213L) mutant , which enhances the strength of interaction with PIP2 ( compared to WT ) , was significantly less sensitive ( IC50 = 9 μM ) . In inside-out patches , continuous application of PIP2 strikingly prevented the mefloquine inhibition . Our results support the idea that mefloquine interferes with PIP2-Kir channels interactions . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . Type B gamma-aminobutyric acid receptors modulate the function of the extracellular Ca2+-sensing receptor and cell differentiation in murine growth plate chondrocytes . Extracellular calcium-sensing receptors ( CaRs ) and metabotropic or type B gamma-aminobutyric acid receptors ( GABA-B-Rs ) , two closely related members of family C of the G protein-coupled receptor superfamily , dimerize in the formation of signaling and membrane-anchored receptor complexes . We tested whether CaRs and two GABA-B-R subunits ( Q96GN5 and R2 ) are expressed in mouse growth plate chondrocytes ( GPCs ) by PCR and immunocytochemistry and whether interactions between these receptors influence the expression and function of the CaR and extracellular Ca(2+)-mediated cell differentiation . Both CaRs and the Q9UBS5 and -R2 were expressed in the same zones of the growth plate and extensively colocalized in intracellular compartments and on the membranes of cultured GPCs . The Q9UBS5 co-immunoprecipitated with the CaR , confirming a physical interaction between the two receptors in GPCs . In vitro knockout of Q9UBS5 genes , using a Cre-lox recombination strategy , blunted the ability of high extracellular Ca(2+) concentration to activate phospholipase C and P27361 /2 , suppressed cell proliferation , and enhanced apoptosis in cultured GPCs . In GPCs , in which the Q9UBS5 was acutely knocked down , there was reduced expression of early chondrocyte markers , aggrecan and type II collagen , and increased expression of the late differentiation markers , type X collagen and osteopontin . These results support the idea that physical interactions between CaRs and GABA-B-R1s modulate the growth and differentiation of GPCs , potentially by altering the function of CaRs . Screening for candidate gene regions in narcolepsy using a microsatellite based approach and pooled DNA . Narcolepsy is a complex sleep disorder characterized by excessive daytime sleepiness and cataplexy . Mutations in genes of the hypocretin ( orexin ) neurotransmitter system cause narcoleptic symptoms in animal models . The absence of hypocretin in the cerebrospinal fluid of human patients is hypothesized to originate from destruction of hypocretinergic cells in the hypothalamus , the cause of which remains unknown . Due to strong HLA association autoimmune models of narcolepsy pathogenesis are still mostly favored . Genetic predisposition factors other than HLA are likely to play a role in causing the disorder . We screened three sets of gene regions ( n=254 ) for association with narcolepsy using a microsatellite based approach and pooled DNA : genes related to immunity , particularly apoptosis ; genes related to regulation of circadian rhythmicity ; genes coding for several factors of neurotransmission . In relation to apoptosis an association was found for the Q99933 gene region . Interestingly , microsatellites representing four genomic regions related to neurotransmission revealed association with narcolepsy : P21964 , P14416 , Q9UBS5 , and P28223 . These results , although exploratory and still to be confirmed in independent samples , support a complex pathogenetic model for narcolepsy , including disturbances of neurotransmission rather than involvement of autoimmunity . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Puerarin alleviates noise-induced hearing loss via affecting PKCγ and GABAB receptor expression . Noise-induced hearing loss ( NIHL ) often results from prolonged exposure to high levels of noise . Our previous study revealed that during the development of NIHL , the expression of protein kinase C γ subunit ( PKCγ ) and GABAB receptor ( GABABR ) was changed within the cochlear nuclear complex ( CNC ) , suggesting that these molecules might be the potential targets for the treatment of NIHL . As an extending study , here we focused on puerarin , a major isoflavonoid extracted from Pueraria lobota , which has been used in the treatment of cardiovascular and cerebrovascular diseases , and investigated whether it could protect against NIHL by acting on PKCγ and GABABR . Transgenic Q99259 -GFP knock-in mice were subjected to the NIHL model and their auditory functions were evaluated by the auditory brainstem response thresholds and distortion product oto-acoustic emission signals . Our results showed that 200mg/kg puerarin treatment ameliorated the thresholds of auditory brainstem response of NIHL mice significantly . Triple immunofluorescence staining and electron microscopy results revealed that GFP-positive neurons in the superficial layers of CNC expressed both PKCγ and Q9UBS5 , and Q99259 -positive terminals contacted PKCγ- or Q9UBS5 -positive neurons . Immunoblotting and RT-PCR results showed that NIHL increased the expression of PKCγ but decreased that of Q9UBS5 and O75899 at both protein and mRNA levels in the CNC . Puerarin significantly attenuated the increased expression of PKCγ but elevated the reduced expression of Q9UBS5 and O75899 after noise exposure . Thus , we provided the first evidence that puerarin ameliorated the auditory functions of NIHL mice , and this effect may be due to its ability to regulate the expression of PKCγ and GABABR . Complex formation with the Type B gamma-aminobutyric acid receptor affects the expression and signal transduction of the extracellular calcium-sensing receptor . Studies with P29320 -293 cells and neurons . We co-immunoprecipitated the Ca(2+)-sensing receptor ( CaR ) and type B gamma-aminobutyric acid receptor ( GABA-B-R ) from human embryonic kidney ( P29320 ) -293 cells expressing these receptors and from brain lysates where both receptors are present . CaRs extensively co-localized with the two subunits of the GABA-B-R ( Q96GN5 and R2 ) in P29320 -293 cell membranes and intracellular organelles . Coexpressing CaRs and GABA-B-R1s in P29320 -293 cells suppressed the total cellular and cell surface expression of CaRs and inhibited phospholipase C activation in response to high extracellular [ Ca(2+) ] ( [Ca(2+)](e) ) . In contrast , coexpressing CaRs and GABA-B-R2s enhanced CaR expression and signaling responses to raising [Ca(2+)](e) . The latter effects of the O75899 on the CaR were blunted by coexpressing the Q9UBS5 . Coexpressing the CaR with Q9UBS5 or R2 enhanced the total cellular and cell surface expression of the Q9UBS5 or R2 , respectively . Studies with truncated CaRs indicated that the N-terminal extracellular domain of the CaR participated in the interaction of the CaR with the Q9UBS5 and R2 . In cultured mouse hippocampal neurons , CaRs co-localized with the Q9UBS5 and R2 . CaRs and GABA-B-R1s also co-immunoprecipitated from brain lysates . The expression of the CaR was increased in lysates from Q9UBS5 knock-out mouse brains and in cultured hippocampal neurons with their Q9UBS5 genes deleted in vitro . Thus , CaRs and GABA-B-R subunits can form heteromeric complexes in cells , and their interactions affect cell surface expression and signaling of CaR , which may contribute to extracellular Ca(2+)-dependent receptor activation in target tissues . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Identification of cell surface targets for HIV-1 therapeutics using genetic screens . Human immunodeficiency virus ( HIV ) drugs designed to interfere with obligatory utilization of certain host cell factors by virus are less likely to encounter development of resistant strains than drugs directed against viral components . Several cellular genes required for productive infection by HIV were identified by the use of genetic suppressor element ( GSE ) technology as potential targets for anti-HIV drug development . Fragmented cDNA libraries from various pools of human peripheral blood mononuclear cells ( PBMC ) were expressed in vitro in human immunodeficiency virus type 1 ( HIV-1 ) -susceptible cell lines and subjected to genetic screens to identify GSEs that interfered with viral replication . After three rounds of selection , more than 15000 GSEs were sequenced , and the cognate genes were identified . The GSEs that inhibited the virus were derived from a diverse set of genes including cell surface receptors , cytokines , signaling proteins , transcription factors , as well as genes with unknown function . Approximately 2.5 % of the identified genes were previously shown to play a role in the HIV-1 life cycle ; this finding supports the biological relevance of the assay . GSEs were derived from the following 12 cell surface proteins : P61073 , CCR4 , P32248 , P20702 , P16070 , Q08722 , P34810 , Q07108 , P04233 , Q99062 , Q9UBS5 , and P20333 . Requirement of some of these genes for viral infection was also investigated by using RNA interference ( RNAi ) technology ; accordingly , 10 genes were implicated in early events of the viral life cycle , before viral DNA synthesis . Thus , these cell surface proteins represent novel targets for the development of therapeutics against HIV-1 infection and AIDS . Differential display RT-PCR reveals genes associated with lithium-induced neuritogenesis in SK-N-MC cells . DB01356 is shown to be neurotrophic and protective against variety of environmental stresses both in vitro as well as in vivo . In view of the wider clinical applications , it is necessary to examine alterations in levels of expression of genes affected by lithium . DB01356 induces neuritogenesis in human neuroblastoma cell line SK-N-MC . Our aim was to elucidate genes involved in lithium-induced neuritogenesis using SK-N-MC cells . The differential display reverse transcriptase polymerase chain reaction ( DD-RT-PCR ) technique was used to study gene expression profiles in SK-N-MC cells undergoing lithium-induced neuritogenesis . Differential expression of genes in control and lithium ( 2.5 mM , 24 h ) -treated cells was compared by display of cDNAs generated by reverse transcription of mRNA followed by PCR using arbitrary primers . Expression of four genes was altered in lithium-treated cells . Real-time PCR was done to confirm the levels of expression of each of these genes using specific primers . DB01356 significantly up-regulated P13591 , a molecule known to stimulate neuritogenesis , occludin , a molecule participating in tight junctions and PKD2 , a molecule known to modulate calcium transport . P01160 32c , a gene whose function is not fully known yet , was found to be down-regulated by lithium . This is the first report demonstrating altered levels of expression of these genes in lithium-induced neuritogenesis and contributes four hitherto unreported candidates possibly involved in the process . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Potent and selective activation of the pancreatic beta-cell type K( DB00171 ) channel by two novel diazoxide analogues . AIMS/HYPOTHESIS : We investigated the pharmacological properties of two novel DB00171 sensitive potassium ( K( DB00171 ) ) channel openers , 6-Chloro-3-isopropylamino-4 H-thieno [ 3,2- e ] -1,2,4-thiadiazine 1,1-dioxide ( NNC 55-0118 ) and 6-chloro-3-(1-methylcyclopropyl)amino-4 H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-dioxide ( NN414 ) , on the cloned cardiac ( Kir6.2/SUR2A ) , smooth muscle ( Kir6.2/SUR2B ) and pancreatic beta cell ( Kir6.2/ Q09428 ) types of K( DB00171 ) channel . METHODS : We studied the effects of these compounds on whole-cell currents through cloned K( DB00171 ) channels expressed in Xenopus oocytes or mammalian cells ( HEK293 ) . We also used inside-out macropatches excised from Xenopus oocytes . RESULTS : In P29320 293 cells , NNC 55-0118 and NN414 activated Kir6.2/ Q09428 currents with EC(50) values of 0.33 micromol/l and 0.45 micromol/l , respectively , compared with that of 31 micro mol/l for diazoxide . Neither compound activated Kir6.2/SUR2A or Kir6.2/SUR2B channels expressed in oocytes , nor did they activate Kir6.2 expressed in the absence of Q09428 . Current activation was dependent on the presence of intracellular MgATP , but was not supported by MgADP . CONCLUSION/INTERPRETATION : Both NNC 55-0118 and NN414 selectively stimulate the pancreatic beta-cell type of K( DB00171 ) channel with a higher potency than diazoxide , by interaction with the Q09428 subunit . The high selectivity and efficacy of the compounds could prove useful for treatment of disease states where inhibition of insulin secretion is beneficial . Benzyl isothiocyanate ( BITC ) inhibits migration and invasion of human gastric cancer AGS cells via suppressing P29323 signal pathways . Metastasis suppressors and associated other regulators of cell motility play a critical initial role in tumor invasion and metastases . Benzyl isothiocyanate ( BITC ) is a hydrolysis compound of glucotropaeolin in dietary cruciferous vegetables . BITC has been found to exhibit prevention of cancers in laboratory animals and might also be chemoprotective in humans . Here , the purpose of this study was to investigate the effects of BITC on cell proliferation , migration , invasion and mitogen-activated protein kinase ( MAPK ) pathways of AGS human gastric cancer cells . Wound healing and Boyden chamber ( migration and invasion ) assays demonstrated that BITC exhibited an inhibitory effect on the abilities of migration and invasion in AGS cancer cells . BITC suppressed cell migration and invasion of AGS cells in a dose-dependent manner . Results from Western blotting indicated that BITC exerted an inhibitory effect on the P27361 /2 , Ras , P62993 , Rho A , P35228 , P35354 for causing the inhibitions of P08253 , -7 and -9 then followed by the inhibitions of invasion and migration of AGS cells in vitro . BITC also promoted O14733 , Q99759 , c-jun , P45983 /2 , P15692 , Sos1 , phosphoinositide 3-kinase ( PI3K ) , PKC , nuclear factor-kappaB ( NF-κB ) p65 in AGS cells . Results from real-time polymerized chain reaction ( PCR ) showed that BITC inhibited the gene expressions of P08253 ,-7 -9 , Q05397 , Q13464 and RhoA after BITC treatment for 24 and 48 hours in AGS cells . Taken together , the finding may provide new mechanisms and functions of BITC , which inhibit migration and invasion of human gastric cancer AGS cells . Immunocytochemical localization of Q9UBS5 receptor subunits in the basolateral amygdala . Gamma-aminobutyric acid B ( GABAB ) receptors ( GBRs ) are G-protein-coupled receptors that mediate a slow , prolonged form of inhibition in the basolateral amygdala ( P00519 ) and other brain areas . Recent studies indicate that this receptor is a heterodimer consisting of Q9UBS5 ( GBR1 ) and O75899 subunits . In the present investigation , antibodies to the Q9UBS5 subunit were used to study the neuronal localization of GBRs in the rat P00519 . GBR immunoreactivity was mainly found in spine-sparse interneurons and astrocytes at the light microscopic level . Very few pyramidal neurons exhibited perikaryal staining . Dual-labeling immunofluorescence analysis indicated that each of the four main subpopulations of interneurons exhibited GBR immunoreactivity . Virtually 100 % of large CCK+ neurons in the basolateral and lateral nuclei were GBR+ . In the basolateral nucleus 72 % of somatostatin ( Q8TE85 ) , 73 % of parvalbumin ( PV ) and 25 % of P01282 positive interneurons were GBR+ . In the lateral nucleus 50 % of somatostatin , 30 % of parvalbumin and 27 % of P01282 positive interneurons were GBR+ . Electron microscopic ( EM ) analysis revealed that most of the light neuropil staining seen at the light microscopic level was due to the staining of dendritic shafts and spines , most of which probably belonged to spiny pyramidal cells . Very few axon terminals ( Ats ) were GBR+ . In summary , this investigation demonstrates that the distal dendrites of pyramidal cells , and varying percentages of each of the four main subpopulations of interneurons in the P00519 , express GBRs . Because previous studies suggest that GBR-mediated inhibition modulates DB01221 -dependent EPSPs in the P00519 , these receptors may play an important role in neuronal plasticity related to emotional learning .

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Genetic polymorphisms , the metabolism of estrogens and breast cancer : a review . Breast cancer is the most common female cancer and the second cause of cancer death in women . Despite recent breakthroughs , much of the etiology of this disease is unknown and the most important risk factor , i.e. , exposure to endogenous and exogenous estrogen throughout life can not explain the heterogeneity of prognosis nor clinical features of patients . Recently , many gene polymorphisms in the metabolism of breast cancer have been described as possible neoplasm etiologic factors . This review is an attempt to summarize the current knowledge about these polymorphisms and to determine new target genes for diagnosis and treatment of the disease . Polymorphisms in the genes P05093 , P11511 , P04798 , P05177 , Q16678 , P22309 , P50225 , 17-hydroxysteroid-dehydrogenase , P21964 , Q86UG4 , P03372 , and Q92731 are described . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . How valid is single nucleotide polymorphism ( SNP ) diagnosis for the individual risk assessment of breast cancer ? The number of reports investigating disease susceptibility based on the carriage of low-penetrance , high-frequency single nucleotide polymorphisms ( SNPs ) has increased in recent years . Evidence is accumulating defining specific individual variations in breast cancer susceptibility . Genetic variations of estradiol and xenobiotics metabolisms as well as genes involved in cell-cycle control have been described as significant contributors to breast cancer susceptibility , with variations depending on ethnic background and co-factors such as smoking and family history of breast cancer . In sum , the highest level of evidence to date linking SNPs and breast cancer comes from nested case-control studies within the prospective Nurses ' Health Study . These data establish seven SNPs - hPRB +331G/A , AR CAG repeat , P11511 (TTTA)10 , P04798 MspI , P11473 FOK1 , P18887 Arg194Trp and O43543 Arg188His - as small but significant risk factors for spontaneous , non-hereditary breast cancer . In addition , meta-analysis of data in the literature establishes the P36897 *6A , HRAS1 , GSTP Ile105Val and P09488 SNPs as low-penetrance genetic risk factors of sporadic breast cancer . The clinical consequences of such a risk elevation may be detailed instruction of the patient as to general measures of breast cancer prevention such as a low-fat diet , optimization of body mass index , physical exercise , avoidance of alcohol and long-term hormone replacement therapy , and participation in a breast cancer screening program between the ages of 50 and 70 years . Specific surgical or drug interventions such as prophylactic mastectomy and oophorectomy or prophylactic intake of tamoxifen are not indicated based on SNP analysis at this time . c-Fos protein as a target of anti-osteoclastogenic action of vitamin D , and synthesis of new analogs . Although active vitamin D drugs have been used for the treatment of osteoporosis , how the vitamin D receptor ( P11473 ) regulates bone cell function remains largely unknown . Using osteoprotegerin-deficient mice , which exhibit severe osteoporosis due to excessive receptor activator of NF-kappaB ligand/receptor activator of NF-kappaB ( O14788 / Q9Y6Q6 ) stimulation , we show herein that oral treatment of these mice with 1alpha,25-dihydroxyvitamin D3 [ DB00136 ] inhibited bone resorption and prevented bone loss , suggesting that P11473 counters O14788 / Q9Y6Q6 signaling . In P09603 -dependent osteoclast precursor cells isolated from mouse bone marrow , DB00136 potently and dose-dependently inhibited their differentiation into multinucleate osteoclasts induced by O14788 . Among signaling molecules downstream of Q9Y6Q6 , DB00136 inhibited the induction of c-Fos protein after O14788 stimulation , and retroviral expression of c-Fos protein abrogated the suppressive effect of DB00136 on osteoclast development . By screening vitamin D analogs based on their c-Fos-suppressing activity , we identified a new analog , named DD281 , that inhibited bone resorption and prevented bone loss in ovariectomized mice , more potently than DB00136 , with similar levels of calcium absorption . Thus , c-Fos protein is an important target of the skeletal action of P11473 -based drugs , and DD281 is a bone-selective analog that may be useful for the treatment of bone diseases with excessive osteoclastic activity . Distinct HDACs regulate the transcriptional response of human cyclin-dependent kinase inhibitor genes to Trichostatin A and 1alpha,25-dihydroxyvitamin D3 . The anti-proliferative effects of histone deacetylase ( HDAC ) inhibitors and 1alpha,25-dihydroxyvitamin D3 [ DB00136 ] converge via the interaction of un-liganded vitamin D receptor ( P11473 ) with co-repressors recruiting multiprotein complexes containing HDACs and via the induction of cyclin-dependent kinase inhibitor ( CDKI ) genes of the INK4 and Cip/Kip family . We investigated the effects of the HDAC inhibitor Trichostatin A ( P32119 ) and DB00136 on the proliferation and CDKI gene expression in malignant and non-malignant mammary epithelial cell lines . P32119 induced the INK4-family genes p18 and p19 , whereas the Cip/Kip family gene P38936 was stimulated by DB00136 . Chromatin immunoprecipitation and RNA inhibition assays showed that the co-repressor NCoR1 and some HDAC family members complexed un-liganded P11473 and repressed the basal level of CDKI genes , but their role in regulating CDKI gene expression by P32119 and DB00136 were contrary . O15379 and HDAC7 attenuated DB00136 -dependent induction of the P38936 gene , for which NCoR1 is essential . In contrast , P32119 -mediated induction of the p18 gene was dependent on O15379 and P56524 , but was opposed by NCoR1 and un-liganded P11473 . This suggests that the attenuation of the response to P32119 by NCoR1 or that to DB00136 by HDACs can be overcome by their combined application achieving maximal induction of anti-proliferative target genes . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . ExoS of Pseudomonas aeruginosa binds to a human Q2M1P5 to induce cytotoxicity in cultured human bronchial epithelial cells . The lungs are a major site of Pseudomonas aeruginosa infection in patients with compromised immune systems . P. aeruginosa secretes a number of toxins by a type III secretion system , and these are important in virulence . One of these toxins , ExoS can induce a cytotoxic effect and is associated with the ability to produce lung damage . ExoS is a bifunctional toxin , with N-terminal P20936 ( P20936 ) activity and a C-terminal ADP ribosyl transferase ( P09874 ) domain . Although these two domains have numerous potential cellular targets , the overall mechanism of ExoS-induced cytotoxicity remains unclear . We carried out a yeast two-hybrid screen using the ExoS truncation mutant ExoSΔ ( residue 1-388 ) , which lacks the 14-3-3 binding site in the P09874 domain , to identify unknown cellular targets associated with ExoS-induced cytotoxicity . We identified the mammalian factor , kinesin family member 7 ( Q2M1P5 ) , which is involved in Hedgehog signaling , as a binding partner for ExoSΔ P06681 . A pull-down assay revealed that ExoS bound to the truncated Q2M1P5 gene encoding the N-terminal domain ( residues 1-109 ) of Q2M1P5 . Yeast two-hybrid analysis showed that the P09874 domain ( residues 234-354 ) of ExoS bound to the truncated Q2M1P5 . Furthermore , exoS gene expression and silencing the expression of Q2M1P5 both caused significant cytotoxicity in cultured human bronchial epithelial cells ( BEAS-2B ) . Taken together , our results suggest that ExoS could induce cytotoxicity in BEAS-2B cells by interacting with Q2M1P5 . 1alpha,25-dihydroxyvitamin D3 and a variety of its natural metabolites transcriptionally repress nuclear-factor-kappaB-mediated interleukin-8 gene expression . Regulation of interleukin-8 ( P10145 ) gene transcription occurs mainly through the sequences -94 to -71 of the 5'-flanking region of the P10145 gene , involving the transcription factors nuclear factor for interleukin-6 ( NF- P05231 ) and nuclear factor kappaB ( NF-kappaB ) . The human melanoma cell line A3 was derived from G-361 cells by stable transfection with an P10145 promoter-luciferase construct containing these sequences . DB00136 ( calcitriol ) repressed P10145 promoter activity induced by tumor necrosis factor-alpha ( P01375 ) by 50 % , compared to 30 % inhibition using dexamethasone , an effect consistent with its effect on P01375 -induced P10145 release and P10145 mRNA levels . A variety of vitamin D metabolites caused the same repressive effect on P10145 promoter activation as calcitriol . However , only those metabolites which were able to transactivate a classical vitamin D response element had the ability to repress P10145 promoter activation , suggesting that this repression is mediated via vitamin D receptor ( P11473 ) . Furthermore , overexpression of P11473 in the parental G-361 cell line enhanced the repressive effect of calcitriol on activation of the P10145 promoter by either P01375 stimulation or overexpression of the NF-kappaB subunit p65 . Electrophoretic mobility shift assays using nuclear extracts from A3 cells showed that calcitriol decreased the abundance of nuclear factors bound to the NF-kappaB binding site of the P10145 promoter and this reduced binding of NF-kappaB proteins presumably contributes to its inhibitory action . Effect of P35354 inhibitors and non-steroidal anti-inflammatory drugs on a mouse fracture model . A randomised , blinded , prospective animal study with 296 male C57BL/6N mice was performed to evaluate the biomechanical , biomolecular , biochemical , and histological impact of anti-inflammatory medications on fracture healing . A reproducible closed tibia fracture was created and stabilised with an intramedullary pin . Animals were randomised to placebo , ketorolac , ibuprofen , celecoxib , or rofecoxib treatment groups with biomechanical and biochemical testing at 4 , 8 , and 12 weeks . A second arm of the study was conducted in which animals were randomised to indomethacin or placebo treatment with biomechanical testing at 12 weeks . Histological and biomolecular studies were performed at 2 weeks on all groups in the first arm of the study . Biomechanical testing consisted of three-point bending evaluating maximum load , energy absorbed to maximum load , and stiffness . Safranin O-Fast Green stain was performed for histology . Biochemical quantifications of chondroitin and dermatan sulphate , hydroxyproline , total protein , and DNA content were performed . P02818 and collagen types II and X were evaluated by in situ hybridisation . Some mechanical differences were seen between ketorolac and placebo at 4 weeks with respect to energy absorbed , but there were no differences in maximum load or stiffness seen between any treatment group and placebo at any time point . Indomethacin , celecoxib , rofecoxib , ibuprofen , and ketorolac did not significantly affect fracture healing in this young murine model . Vitamin D mediates its action in human colon carcinoma cells in a calcium-sensing receptor-dependent manner : downregulates malignant cell behavior and the expression of thymidylate synthase and survivin and promotes cellular sensitivity to DB00544 . Vitamin D ( VD ) protects against colon carcinogenesis by mechanisms not fully understood . We had earlier reported on the similarity in the biologic action of VD and that of the calcium-sensing receptor ( P41180 ) in human colon carcinoma cells . At the molecular level , the P41180 gene contains 2 VD response elements and VD stimulates the expression of P41180 . In this study , we investigated on the relationship between VD action and P41180 function . We determined and compared the action of VD in human colon carcinoma cells ( P35520 , Moser , Caco-2 and HCT116 ) and their P41180 knocked-down counterparts . VD inhibited cellular proliferation , cellular invasion , and anchorage-independent growth and stimulated the expression of P38936 /Waf1 but not in P41180 knocked-down cells . These results demonstrate , for the first time , that the known tumor-suppressive function of VD requires functional P41180 and knocking down P41180 expression abrogated this function of VD . We recently reported that activation of P41180 in human colon carcinoma cells downregulated the expression of thymidylate synthase ( TS ) and survivin and promoted a significant increase in sensitivity to cytotoxic drugs . We now demonstrate , for the first time , that VD suppressed the expression of TS and survivin , TS and survivin gene transcriptional activities and promoted a cytotoxic response to DB00544 in a P41180 -dependent manner . Ectopic expression of wild-type P41180 in colon carcinoma cells also inhibited the expression of TS and survivin and enhanced cellular sensitivity to DB00544 . VD , however , could no longer enhance cellular sensitivity to DB00544 in cells overexpressing P41180 . Genotoxicity and induction of DNA damage responsive genes by food-borne heterocyclic aromatic amines in human hepatoma HepG2 cells . Heterocyclic aromatic amines ( HAAs ) are potential human carcinogens formed in well-done meats and fish . The most abundant are 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine ( PhIP ) , 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline ( MeIQx ) , 2-Amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinoxaline ( 4,8-DiMeIQx ) and 2-Amino-3-methyl-3H-imidazo[4,5-f]quinoline ( IQ ) . HAAs exert genotoxic activity after metabolic transformation by CYP1A enzymes , that is well characterized , however the genomic and intervening responses are not well explored . We have examined cellular and genomic responses of human hepatoma HepG2 cells after 24h exposure to HAAs . Comet assay revealed increase in formation of DNA strand breaks by PhIP , MeIQx and IQ but not 4,8-DiMeIQx , whereas increased formation of micronuclei was not observed . The four HAAs up-regulated expression of genes encoding metabolic enzymes P04798 , P05177 and P22309 and expression of P04637 and its downstream regulated genes P38936 , GADD45α and Q07812 . Consistent with the up-regulation of P38936 and GADD45α the cell-cycle analysis showed arrest in S-phase by PhIP and IQ , and in P55008 -phase by 4,8-DiMeIQx and MeIQx . The results indicate that upon exposure to HAAs the cells respond with the cell-cycle arrest , which enables cells to repair the damage or eliminate them by apoptosis . However , elevated expression of P10415 and down-regulation of Q07812 may indicate that HAAs could suppress apoptosis meaning higher probability of damaged cells to survive and mutate . DB01645 enhancement of respiratory allergen trimellitic anhydride-induced IgE production by adult B6C3F1 mice following in utero and postnatal exposure . The objective of the present study was to determine if exposure to the phytoestrogen genistein ( GEN ) during immune development had any effects on the production of IgE by adult mice following dermal treatment with trimellitic anhydride ( TMA ) , a respiratory allergen . B6C3F1 mice were exposed to GEN either by feeding at 500 ppm or by gavage ( 20 mg/kg ) for varied periods from gestation day ( GD ) 14 to postnatal day ( P01160 ) 84 . In utero exposure to GEN by feeding increased the production of IgE at P01160 84 in male mice but not in female mice . In male mice , continuous exposure to GEN postnatally diminished the in utero exposure-induced enhancement in serum total IgE production . However , continuous exposure to GEN from GD 14 to P01160 84 was required to increase serum total IgE production in female mice . In utero exposure to GEN by gavage increased the production of IgE at P01160 84 in female mice but not in male mice when the mice were maintained on the NIH-07 rodent diet in which a medium level of phytoestrogens was present . The enhancement in IgE production after GEN exposure in females but not in males was associated with decreases in the percentages of P01730 (+)CD25(+) T suppressor cells , and increases in the natural killer ( NK ) cell activity , the basal splenocyte proliferation , the expression of P42081 by B cells , and the production of P60568 and P05112 . Overall , the results demonstrated that GEN differentially modulated the developing immune system in male and female mice , and that more IgE was produced upon exposure to TMA in the adult . Genotyping of patients with sporadic and radiation-associated meningiomas . Ionizing radiation is the most established risk factor for meningioma formation . Our aim was to evaluate the main effect of selected candidate genes on the development of meningioma and their possible interaction with ionizing radiation in the causation of this tumor . The total study population included 440 cases and controls : 150 meningioma patients who were irradiated for tinea capitis in childhood , 129 individuals who were similarly irradiated but did not develop meningioma , 69 meningioma patients with no previous history of irradiation , and 92 asymptomatic population controls . DNA from peripheral blood samples was genotyped for single nucleotide polymorphisms ( SNP ) in 12 genes : P35240 , P18887 , O43542 , P13010 , P18074 , Ki-ras , p16 , cyclin D1 , P60484 , P12830 , P01137 , and P37173 . SNP analysis was done using the MassArray system ( Sequenom , San Diego , CA ) and computerized analysis by SpectroTYPER . Logistic regressions were applied to evaluate main effect of each gene on meningioma formation and interaction between gene and radiation . Intragenic SNPs in the Ki-ras and P18074 genes were associated with meningioma risk ( odds ratio , 1.76 ; 95 % confidence interval , 1.07-2.92 and odds ratio , 1.68 ; 95 % confidence interval , 1.00-2.84 , respectively ) . A significant interaction was found between radiation and cyclin D1 and p16 SNPs ( P for interaction = 0.005 and 0.057 , respectively ) . Our findings suggest that Ki-ras and P18074 SNPs are possible markers for meningioma formation , whereas cyclin D1 and p16 SNPs may be markers of genes that have an inverse effect on the risk to develop meningioma in irradiated and nonirradiated populations . Nuclear factor of activated T cells ( NFAT ) as a molecular target for 1alpha,25-dihydroxyvitamin D3-mediated effects . The molecular basis of the immunomodulatory properties of 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) remains elusive . We demonstrate here that DB00136 -mediated suppressive effects on the inducible expression of cytokine genes in human T cells may , in part , be due to diminished activity of the transcription factor NFAT . The vitamin D3 receptor ( P11473 ) and its heterodimeric partner retinoid X receptor alpha ( RXR alpha ) specifically bound to the distal NFAT site in the human P60568 promoter , and this binding was abolished by mutating unique regions in the NFAT oligonucleotide . In vitro inhibition of NFAT complex formation was noted when P11473 -RXR alpha heterodimers were added to DNA binding reactions containing nuclear extracts from activated B or T cells , whereas in vitro NFkappaB complex formation was not significantly influenced . Furthermore , DB00136 treatment of activated T cells resulted in decreased formation of NFAT complexes detected upon incubation of nuclear extracts from these cells with 32P-labeled probe . Transient expression of both P11473 and RXR alpha , but not of a single component , was capable of inhibiting expression of a NFAT-driven reporter gene in stimulated jurkat cells in a ligand-dependent manner . These results suggest that NFAT plays a crucial role in DB00136 -mediated immunosuppressive activity . Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins . Poly(ADP-ribose) is formed in possibly all multicellular organisms by a familiy of poly(ADP-ribose) polymerases ( PARPs ) . P09874 , the best understood and until recently the only known member of this family , is a DNA damage signal protein catalyzing its automodification with multiple , variably sized ADP-ribose polymers that may contain up to 200 residues and several branching points . Through these polymers , P09874 can interact noncovalently with other proteins and alter their functions . Here we report the discovery of a poly(ADP-ribose)-binding sequence motif in several important DNA damage checkpoint proteins . The 20-amino acid motif contains two conserved regions : ( i ) a cluster rich in basic amino acids and ( ii ) a pattern of hydrophobic amino acids interspersed with basic residues . Using a combination of alanine scanning , polymer blot analysis , and photoaffinity labeling , we have identified poly(ADP-ribose)-binding sites in the following proteins : p53 , P38936 (CIP1/ P38936 ) , xeroderma pigmentosum group A complementing protein , P52701 , P49916 , P18887 , DNA polymerase epsilon , DNA-PK(CS) , P12956 , NF-kappaB , inducible nitric-oxide synthase , caspase-activated DNase , and telomerase . The poly(ADP-ribose)-binding motif was found to overlap with five important functional domains responsible for ( i ) protein-protein interactions , ( ii ) DNA binding , ( iii ) nuclear localization , ( iv ) nuclear export , and ( v ) protein degradation . Thus , PARPs may target specific signal network proteins via poly(ADP-ribose) and regulate their domain functions . Role of efflux pump activity in lapatinib/caelyx combination in breast cancer cell lines . The aim of this study was to investigate , at preclinical level , efflux pump modulation induced by lapatinib , a small-molecule dual inhibitor of the epidermal growth factor receptor ( P00533 ) , in P04626 -negative or P04626 -positive breast cancer cell lines ( SkBr3 and BRC230 ) . We also evaluated the cytotoxic activity and modulation of biomolecular cellular pathways regulated by caelyx and lapatinib , used singly or in combination , at concentrations corresponding to peak plasma level in the two cell lines . DB01259 was active in the P04626 -overexpressing cell line , SkBr3 , but not in BRC230 cell line , which does not express P04626 . Conversely , caelyx exerted a cytotoxic effect on both the cell lines . Simultaneous exposure to lapatinib and caelyx in SkBr3 cell line produced an additive cytotoxic effect with dephosphorylation of P04626 and P00533 , an upregulation of P38936 , and an induction of apoptosis through dephosphorylation of Q92934 and caspase cleavage . In BRC230 , simultaneous treatment induced a synergistic effect that was because of , at least in part , an upregulation of P38936 . DB01259 also blocked efflux pumps , such as the breast cancer resistance protein I by increasing the length of time in which caelyx was present in tumor cell cytoplasm , which led to caspase cleavage , Q92934 dephosphorylation , and apoptosis . Our data indicate that lapatinib used in combination with caelyx is active in P04626 -expressing cells , probably because of lapatinib-induced dephosphorylation of the P04626 - P00533 pathway , and also in non- P04626 -expressing cells , possibly because lapatinib blocks efflux pump activity , increasing the length of time of intracellular exposure to caelyx and thereby increasing its cytotoxic effect . Extracorporeal shock waves stimulate osteoblast activities . The extracorporeal shock wave therapy ( ESWT ) is an extensively applied treatment for musculoskeletal disorders because it promotes bone repair . The aim of this study was to evaluate the direct effect of ESWT on murine osteoblasts to clarify the cellular mechanism that leads to the induction of osteogenesis . Osteoblasts in culture flasks were treated with ESWT pulses ( 500 impulses of 0.05 mJ/mm(2) ) generated by an electromagnetic device . Using western blot analysis 3h after ESWT , an increased expression of Bax was found , indicating a fast pro-apoptotic effect of treatment on some of the osteoblasts . Activation of the cyclin E2/ P24941 is the complex that regulates the P55008 -S transition and is essential for cell proliferation . It was evident 24 to 72h after treatment , indicating a proliferative stimulus . A decreased expression of osteoprotegerin ( O00300 ) and receptor activator NF kappa B ligand ( O14788 ) 24 and 48h after ESW , followed by a later increase of O00300 , paired with a much smaller increase of O14788 , was evident by real-time polymerase chain reaction ( PCR ) . The decreased O14788 / O00300 ratio suggests inhibition of osteoclastogenesis . We can conclude that ESWT induces bone repair through the proliferation and differentiation of osteoblasts and the reduction of their secretion of pro-osteoclastogenic factors . Oxidative stress and cigarette smoke alter chromatin remodeling but differentially regulate NF-kappaB activation and proinflammatory cytokine release in alveolar epithelial cells . Oxidative stress is implicated in lung inflammation due to its effect on proinflammatory gene transcription . Changes in gene transcription depend on chromatin remodeling and the relative activities of histone acetyltransferases ( HATs ) and histone deacetylases ( HDACs ) . Alterations in the nuclear histone acetylation:deacetylation balance may result in uncontrolled transcription of specific proinflammatory genes . We studied the effect of hydrogen peroxide ( H2O2 ) and cigarette smoke condensate ( CSC ) on histone acetylation:deacetylation in human alveolar epithelial cells ( A549 ) . H2O2 and CSC significantly increased acetylation of histone H4 proteins and were associated with decreased HDAC activity and Q92769 levels in A549 cells . Also , the decreased Q92769 activity was due to protein modification by aldehydes and nitric oxide products . Pretreatment of A549 cells with N-acetyl-l-cysteine attenuated the oxidant-mediated reduction in HDAC activity . Treatment of A549 cells with CSC did not cause nuclear factor-kappaB ( NF-kappaB ) activation or expression and release of either interleukin ( IL ) -8 or P05231 . However , H2O2 , tumor necrosis factor-alpha ( P01375 ) , and IL-1beta significantly increased NF-kappaB activation and expression of P10145 compared with control cells . Interestingly , CSC dose dependently inhibited P01375 - and IL-1beta-mediated NF-kappaB activation and P10145 expression . Thus , H2O2 and CSC enhance acetylation of histone proteins and decrease histone deacetylase activity but differentially regulate proinflammatory cytokine release in alveolar epithelial cells . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Molecular-pathological prognostic factors of gastric cancer : a review . Invasion and metastasis are critical determinants of cancer morbidity . Genes and molecules participating in these steps must be regarded as potential prognostic factors . Growth factors and their receptors , cell-cycle regulators , cell-adhesion molecules and matrix-degrading enzymes are those to be used as prognostic factors , including epidermal growth factor ( P01133 ) , P01133 receptor , P21802 , HER-2 , interleukin ( IL ) -8 , vascular endothelial growth factor ( P15692 ) , cyclin E , p27 , P12830 , CD44v6 , matrix metalloproteinase-1 ( P03956 ) , and tissue inhibitor of matrix metalloproteinase-1 ( P01033 ) . Alterations in epigenetics , such as aberrant DNA methylation and histone modification that are , in part , associated with the tumor progression of gastric cancer , can be candidate prognostic factors . The number of methylated genes may serve as a marker of tumor progression . Genetic polymorphism not only affects cancer susceptibility but also influences malignant phenotype ; examples include single-nucleotide polymorphism in the HER-2 and P14780 genes . Comprehensive gene expression analyses are useful to search for novel genes related to invasion and metastasis and potential prognostic factors . Serial analysis of gene expression ( Q9NXZ1 ) has identified several these genes , such as Q12864 , P02649 , P35637 , P02452 , P08123 , Q6UX06 , and Q16674 . Overexpression of Q16674 is found to be associated with poor prognosis . Microarray analysis has great potential for identifying the characteristics of individual cancers , from the view point of gene expression profiles . A combination of these examinations can not only foretell a patient 's prognosis but can also give information directly connected with personalized cancer medicine and prevention . Protein kinases mediate ligand-independent derepression of sumoylated progesterone receptors in breast cancer cells . In advanced breast tumors , protein kinases are upregulated and steroid hormone receptors often function independently of ligand . Herein , we explored mechanisms of ligand-independent progesterone receptor ( PR ) activity . We showed previously that growth factor-induced phosphorylation of PR DB00133 -294 blocks PR Lys-388 sumoylation . SUMO-deficient mutant PR-B ( K388R ) thus provides a model receptor for the study of PR function in the context of high kinase activities . T47D cells stably expressing K388R PR-B exhibited increased ligand-independent proliferation and growth in soft agar relative to cells expressing wt PR-B or phospho-mutant ( sumoylated ) S294A PR-B . Expression of selected PR target genes ( HB- P01133 , P35568 , and P52823 ) was significantly elevated in cells containing desumoylated ( K388R ) PR-B . Basal PR transcriptional activity occurred independently of progestins , was increased by activated P24941 , and attenuated by DB00834 . Notably , ChIP assays demonstrated that K388R PR-B and SRC1 were constitutively recruited to the P52823 promoter in the absence of progestin ; PR Lys-388 sumoylation was required for O15379 recruitment . Knock-down of P52823 inhibited proliferation of cells expressing K388R PR-B . These data suggest a mechanism whereby phosphorylated , and thus desumoylated , PRs mediate increased expression of growth promoting genes . Our data explain why breast cancer models often remain insensitive to progestins , but are growth-inhibited by antiprogestins , and underscore the need to target PR-B and associated kinase activities as part of breast cancer therapy . Evidence that Q13507 is a molecular component of the DB00136 -activated capacitative calcium entry ( CCE ) in muscle and osteoblast cells . In chick skeletal muscle and in rat osteoblast-like cells ( ROS 17/2.8 ) , 1alpha,25-dihydroxy-Vitamin-D(3) [ 1alpha,25(OH)(2)D(3) ] stimulates release of Ca(2+) from inner stores and extracellular cation influx through both voltage-dependent and capacitative Ca(2+) entry ( CCE ) channels . We investigated the involvement of TRPC proteins in CCE induced by 1alpha,25(OH)(2)D(3) . Two fragments were amplified by RT-PCR , exhibiting > 85 % sequence homology with human Q13507 . Northern and Western blots employing Q13507 -probes and anti- Q13507 antibodies , respectively , confirmed endogenous expression of a Q13507 -like protein . Both cell types transfected with anti- Q13507 antisense oligodeoxynucleotides showed reduced CCE and Mn(2+) entry induced by either thapsigargin or 1alpha,25(OH)(2)D(3) . In muscle cells , anti- P11473 antisense inhibited steroid-induced Ca(2+) and Mn(2+) influx and co-immunoprecipitation of Q13507 and P11473 was observed , suggesting an association between both proteins and a functional role of the receptor in 1alpha,25(OH)(2)D(3) activation of CCE . In osteoblasts , two PCR fragments showing high homology with human INAD-like sequences were obtained . Northern blot and antisense functional assays suggested the involvement of the INAD-like protein in CCE regulation by the hormone . Therefore , we propose that an endogenous Q13507 protein mediates 1alpha,25(OH)(2)D(3) modulation of CCE in muscle and osteoblastic cells , which seems to implicate P11473 - Q13507 association and the participation of a INAD-like scaffold protein . Southwest Oncology Group study S0413 : a phase II trial of lapatinib ( GW572016 ) as first-line therapy in patients with advanced or metastatic gastric cancer . BACKGROUND : DB01259 ( GW572016 ) is a dual tyrosine kinase inhibitor of epidermal growth factor receptor ( P00533 ) and human epidermal growth factor receptor 2 ( P04626 /ErbB2 ) , which are reported as overexpressed in 15 % -45 % of gastric cancers , making them potential targets . PATIENTS AND METHODS : The primary objective of this study was to assess response rate . Secondary objectives included overall survival ( OS ) , toxicity , and the relationship of P00533 , ErbB2 , and markers of angiogenesis with clinical outcome . DB01259 was administered to chemonaive metastatic gastric cancer patients at a dose of 1500 mg orally daily for 28 days . RESULTS : The study enrolled 47 patients from February 2005 until May 2006 . Four patients ( 9 % ) had a confirmed partial response ( PR ) , 1 ( 2 % ) had an unconfirmed PR , and 10 ( 23 % ) had stable disease . Median ( 95 % confidence interval ) time to treatment failure was 1.9 ( 1.6-3.1 ) months and OS was 4.8 ( 3.2-7.4 ) months . Significant adverse events : one grade 4 cardiac ischemia/infarction , one grade 4 fatigue , and one grade 4 emesis . One treatment-related death was due to central nervous system ischemia . An exploratory analysis of markers revealed gene expression of P04626 , interleukin ( IL ) -8 and genomic polymorphisms P10145 , and vascular endothelial growth factor correlated with OS . CONCLUSIONS : DB01259 is well tolerated , with modest single-agent activity in advanced/metastatic gastric cancer patients . Potential molecular correlatives were identified which warrant further validation . The calcium-sensing receptor -- a driver of colon cell differentiation . Dietary Ca(2+) reduces colon cell proliferation and carcinogenesis , but it becomes ineffective or even tumor-promoting during carcinogenesis . It appears that Ca(2+) and the colon cell P41180 together brake the massive cell production in normal colon crypts . The rapid proliferation of the transit-amplifying ( TA ) progeny of the colon stem cells at the bases of the crypts is driven by the " Wnt " signaling mechanism that stimulates proliferogenic genes and prevents apoptogenesis . It appears that TA cell cycling stops and terminal differentiation starts when the cells reach a higher level in the crypt where there is enough external Ca(2+) to stimulate the expression of CaSRs , the signals from which stimulate the expression of P12830 . At this point the P25054 ( adenomatous polyposis coli ) protein appears and some of it enters the nucleus . There it removes the apoptogenesis shield and stops the beta-cateninTcf-4 complex from driving further TA cell proliferation by releasing beta-catenin from the nucleus , and delivering it to cytoplasmic APCaxinGSK-3beta complexes for ultimate proteasomal destruction . Cytoplasmic beta-catenin is prevented from returning to the nucleus by destruction in APCaxinGSK-3beta complexes or locked by the emerging P12830 into adherens junctions which link the cell to proliferatively shut-down functioning cells with P25054 -dependent cytoskeletons moving up and out of the crypt . A common first step in colon carcinogenesis is the loss of functional P25054 which results in the retention of proliferogenic nuclear beta-cateninTcf-4 . This drives the eventual appearance of mutation accumulating , apoptosis-resistant clones the proliferation of which can not be inhibited by external Ca(2+) because of P41180 -disabling gene mutations . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . A cardiac pathway of cyclic GMP-independent signaling of guanylyl cyclase A , the receptor for atrial natriuretic peptide . Cardiac atrial natriuretic peptide ( P01160 ) regulates arterial blood pressure , moderates cardiomyocyte growth , and stimulates angiogenesis and metabolism . P01160 binds to the transmembrane guanylyl cyclase ( GC ) receptor , P16066 , to exert its diverse functions . This process involves a cGMP-dependent signaling pathway preventing pathological [Ca(2+)](i) increases in myocytes . In chronic cardiac hypertrophy , however , P01160 levels are markedly increased and P16066 /cGMP responses to P01160 are blunted due to receptor desensitization . Here we show that , in this situation , P01160 binding to P16066 stimulates a unique cGMP-independent signaling pathway in cardiac myocytes , resulting in pathologically elevated intracellular Ca(2+) levels . This pathway involves the activation of Ca(2+)-permeable transient receptor potential canonical 3/6 ( Q13507 / P13671 ) cation channels by P16066 , which forms a stable complex with Q13507 / P13671 channels . Our results indicate that the resulting cation influx activates voltage-dependent L-type Ca(2+) channels and ultimately increases myocyte Ca(2)(+)(i) levels . These observations reveal a dual role of the P01160 / P16066 -signaling pathway in the regulation of cardiac myocyte Ca(2+)(i) homeostasis . Under physiological conditions , activation of a cGMP-dependent pathway moderates the Ca(2+)(i)-enhancing action of hypertrophic factors such as angiotensin II . By contrast , a cGMP-independent pathway predominates under pathophysiological conditions when P16066 is desensitized by high P01160 levels . The concomitant rise in [Ca(2+)](i) might increase the propensity to cardiac hypertrophy and arrhythmias . The biological activities of 1alpha,25-dihydroxyvitamin D3 and its synthetic analog 1alpha,25-dihydroxy-16-ene-vitamin D3 in normal human osteoblastic cells and human osteosarcoma SaOS-2 cells are modulated by 17-beta estradiol and dependent on stage of differentiation . We compared the effects of 1alpha,25-dihydroxyvitamin D3 [ DB00136 ] and its analog , 1alpha,25-dihydroxy-16-ene-vitamin D3 [ 1alpha,25(OH)2-16-ene-D3 ] , as well as their interactions with 17-beta estradiol ( E2 ) on osteoblastic function in our human normal ( HOB ) and osteosarcoma SaOS-2 cell models representing two different stages of differentiation , the more differentiated HOB+DEX cells and SaOS+DEX cells , and the corresponding less differentiated HOB-DEX and SaOS-DEX cells . The differential effects of DB00136 and 1alpha,25(OH)2-16-ene-D3 and the modulation by E2 on ALP activity in HOB-DEX and HOB+DEX cells were small but significant . The most significant effects were seen in SaOS+DEX cells , in which 1alpha,25(OH)2-16-ene-D3 was 100-fold more potent than DB00136 , the maximal enhancement being exerted at 0.1 nM and 10 nM , respectively . E2 enhanced the stimulatory effects of both compounds , with ALP being increased 2-fold at 0.1 nM ( p < 0.001 ) . P02818 ( OC ) production in HOB-DEX cells was stimulated 1.3 to 1.4-fold by DB00136 and 1alpha,25(OH)2-16-ene-D3 at a concentration of 0.01 nM , with E2 inhibiting the effect of 1alpha,25(OH)2-16-ene-D3 . In SaOS-DEX and SaOS+DEX cells , DB00136 and 1alpha,25(OH)2-16-ene-D3 stimulated OC production 1.6-fold at 0.1 nM with E2 slightly enhancing the effect of DB00136 . Western blot analysis of DB00136 receptor ( P11473 ) levels showed that in SaOS+DEX cells , the effect of DB00136 was larger than that of 1alpha,25(OH)2-16-ene-D3 . These results show that 1alpha,25(OH)2-16-ene-D3 is biologically active in human osteoblasts . Prenatal exposure to bisphenol A promotes angiogenesis and alters steroid-mediated responses in the mammary glands of cycling rats . Prenatal exposure to Q03001 disturbs mammary gland histoarchitecture and increases the carcinogenic susceptibility to chemical challenges administered long after Q03001 exposure . Our aim was to assess the effect of prenatal Q03001 exposure on mammary gland angiogenesis and steroid hormone pathways in virgin cycling rats . Pregnant Wistar rats were exposed to either 25 or 250 g/kg/day ( 25 and 250 Q03001 , respectively ) or to vehicle . Female offspring were autopsied on postnatal day ( P01160 ) 50 or 110 . Ovarian steroid serum levels , the expression of steroid receptors and their co-regulators Q9Y6Q9 and Q9Y618 in the mammary gland , and angiogenesis were evaluated . At P01160 50 , all Q03001 -treated animals had lower serum levels of progesterone , while estradiol levels remained unchanged . The higher dose of Q03001 increased mammary ERα and decreased Q9Y6Q9 expression at P01160 50 and P01160 110 . Q9Y618 protein levels were similar among groups at P01160 50 , whereas at P01160 110 , animals exposed to 250 Q03001 showed a lower Q9Y618 expression . Interestingly , in the control and 25 Q03001 groups , Q9Y618 increased from P01160 50 to P01160 110 . At P01160 50 , an increased vascular area associated with higher P15692 expression was observed in the 250 Q03001 -treated rats . At P01160 110 , the vascular area was still increased , but P15692 expression was similar to that of control rats . The present results demonstrate that prenatal exposure to Q03001 alters the endocrine environment of the mammary gland and its angiogenic process . Increased angiogenesis and altered steroid hormone signals could explain the higher frequency of pre-neoplastic lesions found later in life . This article is part of a Special Issue entitled ' Endocrine disruptors ' . Recombinant human prothrombin kringle-2 induces bovine capillary endothelial cell cycle arrest at G0- P55008 phase through inhibition of cyclin D1/ P11802 complex : modulation of reactive oxygen species generation and up-regulation of cyclin-dependent kinase inhibitors . P00734 is a plasma glycoprotein involved in blood coagulation and , as we have previously reported , prothrombin kringles inhibit BCE ( bovine capillary endothelial ) cell proliferation . To reveal the mechanism , we investigated the influence of rk-2 ( recombinant human prothrombin kringle-2 ) on the BCE cell cycle progression and ROS ( reactive oxygen species ) generation using FACS ( fluorescence-activated cell sorter ) analysis . Cell cycle analysis showed a decrease of G(1) phase cells in cells treated with P09038 ( basic fibroblast growth factor ) and an increase in cells treated with rk-2 , as compared with the control cells . But , the portion of the S phase was reversed . In Western blot analysis , P09038 induced cytoplasmic translocation of P38936 (Waf1/Cip1) and p27(Kip1) and phosphorylation of p27(Kip1) but rk-2 treatment inhibited translocation of P38936 (Waf1/Cip1) and p27(Kip1) from nucleus to cytoplasm and phosphorylation of p27(Kip1) . Also , rk-2 induced up-regulation of p53 and nuclear P38936 (Waf1/Cip1) and inhibited the cyclin D1/ P11802 ( cyclin-dependent kinase 4 ) complex . The ROS level of rk-2-treated BCE cells was increased 2-fold when compared with the control , but treatment with Q9C000 ( N-Acetyl-L : -cysteine ) , an anti-oxidant , decreased ROS generation about 55 % as compared with the rk-2 treatment . Q9C000 treatment also restored cell cycle progression inhibited by rk-2 and down-regulated p53 and nuclear P38936 (Waf1/Cip1) expression induced by rk-2.These data suggest that rk-2 induces the BCE cell cycle arrest at G(0)-G(1) phase through inhibition of the cyclin D1/ P11802 complex caused by increase of ROS generation and nuclear cyclin-dependent kinase inhibitors . 1Alpha,25(OH)2D3-induced transrepression by vitamin D receptor through E-box-type elements in the human parathyroid hormone gene promoter . Although transactivation by the liganded vitamin D receptor ( P11473 ) is well described at the molecular level , the precise molecular mechanism of negative regulation by the liganded P11473 remains to be elucidated . We have previously reported a novel class of negative vitamin D response element ( nVDRE ) called 1alphanVDRE in the human 25(OH)D31alpha-hydroxylase [ 1alpha(OH)ase ] gene by DB00136 -bound P11473 . This element was composed of two E-box-type motifs that bound to VDIR for transactivation , which was attenuated by liganded P11473 . Here , we explore the possible functions of VDIR and E-box motifs in the human ( h ) PTH and hPTHrP gene promoters . Functional mapping of the DB05829 and hPTHrP promoters identified E-box-type elements acting as nVDREs in both the DB05829 promoter ( hPTHnVDRE ; -87 to -60 bp ) and in the hPTHrP promoter ( hPTHrPnVDRE ; -850 to -600 bp ; -463 to -104 bp ) in a mouse renal tubule cell line . The hPTHnVDRE alone was enough to direct ligand-induced transrepression mediated through P11473 /retinoid X receptor and VDIR . Direct DNA binding of hPTHnVDRE to VDIR , but not P11473 /retinoid X receptor , was observed and ligand-induced transrepression was coupled with recruitment of P11473 and histone deacetylase 2 ( Q92769 ) to the DB05829 promoter . These results suggest that negative regulation of the DB05829 gene by liganded P11473 is mediated by VDIR directly binding to the E-box-type nVDRE at the promoter , together with recruitment of an HDAC corepressor for ligand-induced transrepression . An essential role of the CAAT/enhancer binding protein-alpha in the vitamin D-induced expression of the human steroid/bile acid-sulfotransferase ( Q06520 ) . The vitamin D receptor ( P11473 ) regulates steroid and drug metabolism by inducing the genes encoding phase I and phase II enzymes . Q06520 is a liver- and intestine-expressed sulfo-conjugating enzyme that converts the alcohol-OH of neutral steroids , bile acids , and drugs to water-soluble sulfated metabolites . DB00136 [ 1,25-(OH)2D3 ] induces Q06520 gene transcription after the recruitment of P11473 to the vitamin D-responsive chromatin region of Q06520 . A composite element in human Q06520 directs the 1,25-(OH)2D3-mediated induction of natural and heterologous promoters . This element combines a P11473 /retinoid X receptor-alpha-binding site [ vitamin D response element ( VDRE ) ] , which is an imperfect inverted repeat 2 of AGCTCA , and a CAAT/enhancer binding protein ( C/EBP ) -binding site located 9 bp downstream to VDRE . The binding sites were identified by EMSA , antibody supershift , and deoxyribonuclease I footprinting . C/EBP-alpha at the composite element plays an essential role in the P11473 regulation of Q06520 , because 1 ) induction was lost for promoters with inactivating mutations at the VDRE or C/EBP element ; 2 ) Q06520 induction by 1,25-(OH)2D3 in C/EBP-alpha-deficient cells required the expression of cotransfected C/EBP-alpha ; and 3 ) C/EBP-beta did not substitute for C/EBP-alpha in this regulation . P11473 and C/EBP-alpha were recruited concurrently to the composite element along with the coactivators p300 , steroid receptor coactivator 1 ( Q15788 ) , and P12931 -2 , but not Q9Y6Q9 . P11473 and C/EBP-alpha associated endogenously as a DNA-dependent , coimmunoprecipitable complex , which was detected at a markedly higher level in 1,25-(OH)2D3-treated cells . These results provide the first example of the essential role of the interaction in cis between C/EBP-alpha and P11473 in directing 1,25-(OH)2D3-induced expression of a P11473 target gene . Antagonistic action of novel 1alpha,25-dihydroxyvitamin D3-26 , 23-lactone analogs on differentiation of human leukemia cells ( HL-60 ) induced by 1alpha,25-dihydroxyvitamin D3 . We examined the effects of two novel 1alpha,25-dihydroxyvitamin D3-26,23-lactone ( 1alpha,25-lactone ) analogues on human promyelocytic leukemia cell ( HL-60 ) differentiation using the evaluation system of the vitamin D nuclear receptor ( P11473 ) /vitamin D-responsive element ( DRE ) -mediated genomic action stimulated by 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) and its analogues . We found that the 1alpha,25-lactone analogues ( 23S ) -25-dehydro-1alpha-hydroxyvitamin-D3-26,23-lactone ( TEI-9647 ) , and ( 23R ) -25-dehydro-1alpha-hydroxyvitamin-D3-26,23-lactone ( TEI-9648 ) bound much more strongly to the P11473 than the natural ( 23S , 25R ) - DB00136 -26,23-lactone , but did not induce cell differentiation even at high concentrations ( 10(-6) M ) . Intriguingly , the differentiation of HL-60 cells induced by DB00136 was inhibited by either TEI-9647 or TEI-9648 but not by the natural lactone . In contrast , retinoic acid or 12-O-tetradecanoylphorbol-13-acetate-induced HL-60 cell differentiation was not blocked by TEI-9647 or TEI-9648 . In separate studies , TEI-9647 ( 10(-7) M ) was found to be an effective antagonist of both DB00136 ( 10(-8) M ) mediated induction of P38936 ( P38936 , CIP1 ) in HL-60 cells and activation of the luciferase reporter assay in COS-7 cells transfected with cDNA containing the DRE of the rat DB00146 -24-hydroxylase gene and cDNA of the human P11473 . Collectively the results strongly suggest that our novel 1alpha,25-lactone analogues , TEI-9647 and TEI-9648 , are specific antagonists of 1alpha , 25(OH)2D3 action , specifically P11473 /DRE-mediated genomic action . As such , they represent the first examples of antagonists , which act on the nuclear P11473 . DB00136 inhibits transcriptional potential of nuclear factor kappa B in breast cancer cells . 1alpha,25-Dihydroxyvitamin D(3) ( VD(3) ) , the biologically active form of vitamin D , may have either pro- or anti-inflammatory activities because of its diverse actions on nuclear factor kappa B ( NF-kappaB ) . Previous studies indicated that VD(3) can either activate or inhibit NF-kappaB via Akt-induced I kappaB alpha phosphorylation and increase in I kappaB alpha synthesis respectively . At present , the relevant contribution of each mechanism has not been fully explored . We observed a VD(3)-mediated NF-kappaB inhibitory effect in vitamin D receptor ( P11473 ) -positive MCF-7 breast cancer cells . We showed that VD(3) induced P11473 -dependent I kappaB alpha expression but still able to lead on transient NF-kappaB p65 nuclear translocation through Akt-induced I kappaB alpha phosphorylation . Upon TNFalpha stimulation , VD(3) was not capable to inhibit I kappaB alpha degradation , p65 nuclear translocation and p65/p50-DNA binding . Here , we found that VD(3) strongly repressed p65 transactivation in MCF-7 cells using Gal4-p65 chimeras system . P11473 was required for the VD(3)-mediated transrepression and mutations in P11473 affected its suppressive ability . We also demonstrated that neither inhibition of p65 phosphorylation nor acetylation was responsible for the transrepression . In fact , we found that treatment of MCF-7 cells with histone deacetylase inhibitors abrogated VD(3)-induced p65 transrepression . In addition , knockdown of two nuclear corepressors O15379 and Q9Y618 relieved p65 transactivation and particular TNFalpha-triggered gene expression . In conclusion , the reduction of gene activation by VD(3) in breast cancer cells was caused by the interference of the transactivation potential of NF-kappaB p65 subunit . Our studies provide a scientific background for rational use of vitamin D in the prevention and treatment of inflammatory diseases . DB00203 enhances neurogenesis and oligodendrogenesis in ischemic brain of middle-aged mouse . Adult neural stem cells give rise to neurons , oligodendrocytes and astrocytes . Aging reduces neural stem cells . Using an inducible nestin-CreER( P24752 )/R26R-yellow fluorescent protein ( YFP ) mouse , we investigated the effect of DB00203 , a phosphodiesterase type 5 ( O76074 ) inhibitor , on nestin lineage neural stem cells and their progeny in the ischemic brain of the middle-aged mouse . We showed that focal cerebral ischemia induced nestin lineage neural stem cells in the subventricular zone ( SVZ ) of the lateral ventricles and nestin expressing NeuN positive neurons and adenomatous polyposis coli ( P25054 ) positive mature oligodendrocytes in the ischemic striatum and corpus callosum in the aged mouse . Treatment of the ischemic middle-aged mouse with DB00203 increased nestin expressing neural stem cells , mature neurons , and oligodendrocytes by 33 , 75 , and 30 % , respectively , in the ischemic brain . These data indicate that DB00203 amplifies nestin expressing neural stem cells and their neuronal and oligodendrocyte progeny in the ischemic brain of the middle-aged mouse . Vitamin D regulates the gut microbiome and protects mice from dextran sodium sulfate-induced colitis . The active form of vitamin D [ DB00136 , 1,25(OH)2D3 ] and the vitamin D receptor ( P11473 ) regulate susceptibility to experimental colitis . The effect of the bacterial microflora on the susceptibility of C57BL/6 mice to dextran sodium sulfate-induced colitis was determined . Mice that can not produce 1,25(OH)2D3 [ Cyp27b1 ( Cyp ) knockout ( KO ) ] , P11473 KO as well as their wild-type littermates were used . Cyp KO and P11473 KO mice had more bacteria from the Bacteroidetes and Proteobacteria phyla and fewer bacteria from the Firmicutes and Deferribacteres phyla in the feces compared with wild-type . In particular , there were more beneficial bacteria , including the Lactobacillaceae and Lachnospiraceae families , in feces from Cyp KO and P11473 KO mice than in feces from wild-type . Helicobacteraceae family member numbers were elevated in Cyp KO compared with wild-type mice . Depletion of the gut bacterial flora using antibiotics protected mice from colitis . 1,25(OH)2D3 treatment ( 1.25 μg/100 g diet ) of Cyp KO mice decreased colitis severity and reduced the numbers of Helicobacteraceae in the feces compared with the numbers in the feces of untreated Cyp KO mice . The mechanisms by which the dysbiosis occurs in P11473 KO and Cyp KO mice included lower expression of P12830 on gut epithelial and immune cells and fewer tolerogenic dendritic cells that resulted in more gut inflammation in P11473 and Cyp KO mice compared with wild-type mice . Increased host inflammation has been shown to provide pathogens with substrates to out-compete more beneficial bacterial species . Our data demonstrate that vitamin D regulates the gut microbiome and that 1,25(OH)2D3 or P11473 deficiency results in dysbiosis , leading to greater susceptibility to injury in the gut . A concise and efficient route to 2alpha-(omega-hydroxyalkoxy)-1alpha,25-dihydroxyvi tam in D3 : remarkably high affinity to vitamin D receptor . [ reaction : see text ] A convenient and potentially valuable synthetic approach to the novel 2alpha-functionalized 1alpha,25-dihydroxyvitamin D3 [ DB00136 ] derivatives ( 1a-c ) , which are the P06681 -epimer of ED-71 and its analogues , has been developed . The C2alpha-modified ring A precursors ( 1,7-enynes 16 , n = 0 , 1 , and 2 ) were constructed stereoselectively starting from D-glucose in high yield . In the synthesized 2alpha-(omega-hydroxyalkoxy)- DB00136 derivatives , 1a and 1b showed a greater binding affinity to vitamin D receptor ( P11473 ) , up to 1.8 times that of the native hormone . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Regulation of the human P38936 (waf1/cip1) gene promoter via multiple binding sites for p53 and the vitamin D3 receptor . The main regulator of the human tumor suppresser gene P38936 (waf1/cip1) is the transcription factor p53 , but more recently it has been suggested to be a primary anti-proliferative target for the nuclear receptor P11473 in the presence of its ligand 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) . To identify P11473 responding regions , we analyzed 20 overlapping regions covering the first 7.1 kb of the P38936 (waf1/cip1) promoter in MCF-7 human breast cancer cells using chromatin immuno-precipitation assays ( ChIP ) with antibodies against p53 and P11473 . We confirmed two known p53 binding regions at approximate positions -1400 and -2300 and identified a novel site at position -4500 . In addition , we found three P11473 -associated promoter regions at positions -2300 , -4500 and -6900 , i.e. two regions showed binding for both p53 and P11473 . In silico screening and in vitro binding assays using recombinant and in vitro translated proteins identified five p53 binding sites within the three p53-positive promoter regions and also five DB00136 response elements within the three P11473 -positive regions . Reporter gene assays confirmed the expected responsiveness of the respective promoter regions to the p53 inducer 5-fluorouracil and DB00136 . Moreover , re-ChIP assays confirmed the functionality of the three DB00136 -reponsive promoter regions by monitoring simultaneous occupancy of P11473 with the co-activator proteins CBP , Q15788 and Q15648 . Taken together , we demonstrated that the human P38936 ( ( waf1/cip1 ) ) gene is a primary DB00136 -responding gene with at least three P11473 binding promoter regions , in two of which also p53 co-localizes . Recruitment and subnuclear distribution of the regulatory machinery during 1alpha,25-dihydroxy vitamin D3-mediated transcriptional upregulation in osteoblasts . The architectural organization of the genome and regulatory proteins within the nucleus supports gene expression in a physiologically regulated manner . In osteoblastic cells ligand activation induces a nuclear punctate distribution of the 1alpha,25-dihydroxy vitamin D3 ( DB00136 ) receptor ( P11473 ) and promotes its interaction with transcriptional coactivators such as Q15788 , NCoA-62/Skip , and Q15648 . Here , we discuss evidence demonstrating that in osteoblastic cells P11473 binds to the nuclear matrix fraction in a DB00136 -dependent manner . This interaction occurs rapidly after exposure to DB00136 and does not require a functional P11473 DNA binding domain . The nuclear matrix-bound P11473 molecules colocalize with the also nuclear matrix-associated coactivator Q15648 . We propose a model where the rapid association of P11473 with the nuclear matrix fraction represents an event that follows DB00136 -dependent nuclear localization of P11473 , but that precedes DB00136 -dependent transcriptional upregulation at target genes . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . Effects of interleukin-2 ( P60568 ) on human plasma lipid , lipoprotein , and P02741 . Six patients with confirmed malignant disease received four consecutive weekly cycles of human recombinant interleukin-2 ( P60568 ) 4 days/week , continuous iv. infusion , 3 X 10(6) U/m2/day . Plasma cholesterol decreased a mean of 7 % within 24 hours after P60568 infusion and decreased by 33 % within 4 days . Plasma cholesterol was significantly lower than baseline concentration by day 21 ( -21 % ) , and day 25 ( -41 % ) was significantly lower than day 21 . Decreased plasma cholesterol was the result of decreased HDL and LDL cholesterol concentrations . Plasma triglyceride demonstrated a mean increase of 46 % after 4 days of therapy and remained greater than baseline concentrations at all time points analyzed . Apolipoprotein AI and AII decreased concomitantly with HDL-cholesterol concentrations , whereas apolipoprotein B after an initial mean decrease of 17 % during the first cycle was not significantly different from baseline during the fourth cycle . P02649 and P08519 were not significantly affected by P60568 treatment . Plasma P02741 ( CRP ) increased by 79 % within 24 hours of therapy , increased by 254 % on day 4 , then decreased to baseline concentrations by day 21 after 3 days off of P60568 . Day 25 CRP was elevated compared to both baseline and day 21 concentrations . P60568 induced plasma lipoprotein changes may be due in part to the induction of interferon gamma . Src and P61073 are involved in the invasiveness of breast cancer cells with acquired resistance to lapatinib . DB01259 is a dual P00533 and ErbB-2 tyrosine kinase inhibitor that has significantly improved the clinical outcome of ErbB-2-overexpressing breast cancer patients . However , patients inexorably develop mechanisms of resistance that limit the efficacy of the drug . In order to identify potential targets for therapeutic intervention in lapatinib-resistant patients , we isolated , from ErbB-2-overexpressing SK-Br-3 breast cancer cells , the SK-Br-3 Lap-R-resistant subclone , which is able to routinely grow in 1 µM lapatinib . Resistant cells have a more aggressive phenotype compared with parental cells , as they show a higher ability to invade through a matrigel-coated membrane . DB01259 -resistant cells have an increased Src kinase activity and persistent levels of activation of P27361 /2 and AKT compared with parental cells . Treatment with the Src inhibitor saracatinib in combination with lapatinib reduces AKT and P27361 /2 phosphorylation and restores the sensitivity of resistant cells to lapatinib . SK-Br-3 Lap-R cells also show levels of expression of P61073 that are higher compared with parental cells and are not affected by Src inhibition . Treatment with saracatinib or a specific P61073 antibody reduces the invasive ability of SK-Br-3 Lap-R cells , with the two drugs showing cooperative effects . Finally , blockade of Src signaling significantly increases P50591 -induced cell death in SK-Br-3 Lap-R cells . Taken together , our results demonstrate that breast cancer cells with acquired resistance to lapatinib have a more aggressive phenotype compared with their parental counterpart , and that Src signaling and P61073 play an important role in this phenomenon , thus representing potential targets for therapeutic intervention in lapatinib-resistant breast cancer patients . Contemporary management of metastatic castration-resistant prostate cancer . PURPOSE OF REVIEW : This review will describe the contemporary management of castration-resistant prostate cancer in the context of multiple recent advances . RECENT FINDINGS : Two novel agents have been added to the therapeutic armamentarium including cabazitaxel and sipuleucel-T . DB06772 , a novel taxane extended survival in men with progressive metastatic CRPC following conventional docetaxel-based chemotherapy . Sipuleucel-T , an autologous dendritic cell based vaccine extended survival in men with relatively asymptomatic metastatic CRPC without visceral metastasis . A third agent , abiraterone acetate , an orally administered P05093 inhibitor , which suppresses androgen synthesis has been preliminarily reported to significantly prolong survival following prior docetaxel and approval by regulatory agencies is anticipated . Baseline and early changes in circulating tumor cells appear useful as a prognostic factor . Additionally , data demonstrated the superiority of denosumab , a Q9Y6Q6 -ligand antagonist , compared to zoledronic acid in the prevention of skeletal related events in men with bone metastases . SUMMARY : DB06772 , sipuleucel-T and denosumab were approved in 2010 by regulatory agencies in the USA for men with metastatic CRPC , and approval of abiraterone acetate is anticipated based on the results of a phase III trial . The evaluation of circulating tumor cells assists in determining prognosis , although its utility for clinical decision-making entails validation . 1,25-Dihydroxycholecalciferol enhances butyrate-induced P38936 (Waf1/Cip1) expression . Butyrate , a short-chain fatty acid produced in the colon , as well as its prodrug tributyrin , reduce proliferation and increase differentiation of colon cancer cells . P38936 (Waf1/Cip1) and p27(Kip1) are negative regulators of cell cycle and are thought to have a key function in the differentiation of various cell lines . We studied the effects of butyrate on differentiation , P11473 expression , as well as on P38936 (Waf1/Cip1) and p27(Kip1) expression in human colon cancer cells ( Caco-2 ) . Butyrate induced cell differentiation , which was further enhanced after addition of DB00136 . Synergistic effect of butyrate and dihydroxycholecalciferol in Caco-2 cells was due to butyrate-induced overexpression of P11473 . While butyrate as well as dihydroxycholecalciferol increased P38936 (Waf1/Cip1) and p27(Kip1) expression , in contrast combined exposure of butyrate and dihydroxycholecalciferol resulted in a synergistic amplification of P38936 (Waf1/Cip1) , but not of p27(Kip1) expression . These data imply that butyrate selectively increases P38936 (Waf1/Cip1) expression via upregulation of P11473 in Caco-2 cells . Antisense inhibition of vitamin D receptor expression induces apoptosis in monoblastoid U937 cells . The active vitamin D3 metabolite DB00136 ( 1,25(OH)2D3 ) acts as an antiproliferative and differentiating agent for the monoblastoid cell line U937 and as an important immunologic mediator implicated particularly in the function of cells belonging to the monocyte/macrophage lineage . These effects are controlled by the vitamin D receptor ( P11473 ) , a member of the steroid hormone receptor family . The objective of this study was to develop U937 transfectants expressing antisense P11473 mRNA , and to use these to examine the role of 1,25(OH)2D3- P11473 interaction in this lineage . A 2-kb P11473 cDNA insert ( including the complete P11473 coding region ) was cloned in an antisense orientation into the EBV episomal vector pMEP4 under the control of an inducible promoter and transfected into U937 . The resultant cell line , DH42 , was hygromycin resistant , contained P11473 cDNA , expressed fewer VDRs than controls , and showed a substantial decrease in antiproliferative response to 1,25(OH)2D3 . However , 1,25(OH)2D3 increased the number of cells expressing macrophage cell surface Ags , including P08571 and CD11b . A subpopulation of smaller cells did not express the differentiation markers after cadmium stimulation . Cell cycle analysis showed shifts in the distribution of cells from P55008 to S phase , which were more pronounced after cadmium treatment . A considerable proportion of cells were outside the cycle and DNA fragmentation confirmed apoptosis . Thus , the functional outcome of the P11473 antisense transfection suggests that in the myelomonocytic lineage , P11473 expression may act as a protective mechanism against programmed cell death . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . DB00136 induces nitric oxide production in cultured endothelial cells . BACKGROUND : Recently , DB00136 ( vitD ) has received increasing interest for its effects on many tissues and organs other than bone . A number of experimental studies have shown that vitD may have an important role in modifying risk for cardiovascular disease . AIMS : This study was planned to test the effects of vitD on endothelial nitric oxide ( NO ) production and to study the intracellular pathways leading to NO release . METHODS : In human umbilical vein endothelial cells ( HUVEC ) cultures the effects of vitD on NO production and p38 , Akt , P29323 and P29474 phosphorylations were examined in absence or in presence of the NO synthase inhibitor L-NAME and protein kinases specific inhibitors SB203580 , wortmannin and UO126 . RESULTS : VitD caused a concentration-dependent increase in NO production . The maximum effect was observed at a concentration of 1 nM and the optimal time of stimulation was 1 min . Effects induced by vitD were abolished by L-NAME and by pre-treatment with protein kinases inhibitors . To verify the effective involvement of vitD receptor ( P11473 ) in the action mechanism of vitD , experiments were repeated in presence of the specific P11473 ligands ZK159222 and ZK191784 . CONCLUSIONS : The results of this study demonstrate that vitD can induce a significant increase in endothelial NO production . VitD interaction with P11473 caused the phosphorylation of p38 , AKT and P29323 leading to P29474 activation . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . [ Gene polymorphism of the vitamin D receptor , vitamin D-binding protein and calcium-sensing receptor in respect of calcium-phosphate disturbances in chronic dialysis patients ] . Dialysed patients suffering from chronic kidney disease ( CKD ) show varied levels of concentration of parathyroid hormone ( PTH ) in the blood . One of the factors in charge of regulating levels of PTH concentration is DB00136 [ 1,25-(OH)2D3 ] . Its deficiency in advanced stages of CKD is common . Vitamin D supplementation is not always effective in reaching levels of PTH concentration recommended by KDIGO for the dialysed patients . That suggests , among other things , disturbances in 1,25-(OH)2D3 , reaching its place of target effect and having the desired final result . Disturbances of vitamin D target pathway can be genetically conditioned , hence the aim of this paper is to describe the distribution of polymorphic variants of vitamin D-binding protein gene ( VDBP ) , vitamin D receptor gene ( P11473 ) and gene of the calcium-sensing receptor ( P41180 ) with respect to PTH concentrations in serum and response to cinacalcet treatment in patients with secondary hyperparathyroidism in view of the differences in demographical , clinical and laboratory data of the dialysed patients . Effects of FK228 , a novel histone deacetylase inhibitor , on human lymphoma U-937 cells in vitro and in vivo . FK228 [ ( E ) -(1S,4S,10S,21R)-7- [ ( Z ) -ethylidene ] -4,21-diisopropyl-2-oxa-12,13-dithia-5,8,20,23-tetraazabicyclo-[8,7,6]-tricos-16-ene-3,6,9,19,22-pentanone ; FR901228 , depsipeptide ] is a novel histone deacetylase inhibitor that shows therapeutic efficacy in Phase I trials of patients with malignant lymphoma . However , its mechanism of action has not been characterized . In this study , we examined the in vitro and in vivo effects of FK228 on human lymphoma U-937 cells . FK228 very strongly inhibited the growth of U-937 cells with an IC(50) value of 5.92 nM . In a scid mouse lymphoma model , mice treated with FK228 once or twice a week survived longer than control mice , with median survival times of 30.5 ( 0.56 mg/kg ) and 33 days ( 0.32 mg/kg ) , respectively ( vs. 20 days in control mice ) . Remarkably , 2 out of 12 mice treated with FK228 ( 0.56 mg/kg once or twice a week ) survived past the observation period of 60 days . The apoptotic population of U-937 cells time-dependently increased to 37.7 % after 48 hr of treatment with FK228 . In addition , FK228 induced P55008 and G2/M arrest and the differentiation of U-937 cells to the CD11b(+)/ P08571 (+) phenotype . Expression of P38936 ( P38936 /Cip1) and gelsolin mRNA increased up to 654- and 152-fold , respectively , after 24hr of treatment with FK228 . FK228 caused histone acetylation in P38936 ( P38936 /Cip1) promoter regions , including the Sp1-binding sites . In conclusion , ( i ) FK228 prolonged the survival time of scid mice in a lymphoma model , and ( ii ) the beneficial effects of FK228 on human lymphoma may be exerted through the induction of apoptosis , cell cycle arrest , and differentiation via the modulation of gene expression by histone acetylation . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . NSA9 , a human prothrombin kringle-2-derived peptide , acts as an inhibitor of kringle-2-induced activation in EOC2 microglia . In neurodegenerative diseases , such as Alzheimer 's and Parkinson 's , microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds . P00734 and kringle-2 increase levels of NO and the mRNA expression of P35228 , IL-1beta , and P01375 in microglial cells . In contrast , the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1beta , P01375 , and P05231 in LPS-activated EOC2 microglia . In this study , we investigated the anti-inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia . Treatment with 20-100 muM of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation . NO production induced by MAPKs and NF-kappaB was similarly reduced by inhibitors of P29323 ( PD98059 ) , p38 ( SB203580 ) , NF-kappaB ( DB06151 ) , and NSA9 . These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2 . Caveolae and caveolin-1 are implicated in 1alpha,25(OH)2-vitamin D3-dependent modulation of Src , MAPK cascades and P11473 localization in skeletal muscle cells . We previously reported that DB00136 induces non-transcriptional rapid responses through activation of MAPKs in C2C12 skeletal muscle cells . However , there is little information on the molecular mechanism underlying the initiation of DB00136 signaling through this pathway . Plasma membrane components have been involved in some non-genomic effects . In this work , we investigated the role of caveolae and caveolin-1 ( cav-1 ) in DB00136 -stimulation of c-Src and MAPKs . When proliferating cells were pretreated with methyl beta cyclodextrin ( MbetaCD ) , a caveolae disrupting agent , under conditions in which cell morphology is not affected and no signs of apoptosis are observed , DB00136 -dependent activation of P27361 /2 , p38 MAPK and c-Src was suppressed . Similar results were obtained by siRNA technology whereby silencing of cav-1 expression abolished activation of c-Src and MAPKs induced by the hormone . By confocal immunocytochemistry it was observed that cav-1 colocalizes with c-Src in the periplasma membrane zone at basal conditions . Hormone treatment disrupted the colocalization of these proteins and redistributed them into cytoplasm and nucleus . Co-immunoprecipitation assays corroborated these observations . Changes in P11473 localization after DB00136 exposure were also investigated . Confocal microscopy images showed that the hormone induces P11473 translocation to the plasma membrane , and this effect is abolished by MbetaCD . Altogether , these data suggest that caveolae is involved upstream in c-Src-MAPKs activation by DB00136 and that P11473 and cav-1 participate in the rapid signaling elicited by the hormone .

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interacts_with DB08868?

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DB03203 -1-phosphate induces thrombin receptor Q96RI0 expression to enhance cell migration and P35354 formation in human monocytes . Thrombin is not only a central factor in blood coagulation but also stimulates inflammatory processes , including monocyte responses , via activation of PARs . The signaling lipid Q14703 is a major determinant of monocyte function . Here , we established an interaction between Q14703 and human monocyte responses to thrombin . Q14703 induced P25116 and Q96RI0 mRNA and total protein expression in human monocytes and U937 cells in a concentration ( 0.1-10 μM ) - and time ( 1-24 h ) -dependent manner , respectively . However , only Q96RI0 cell-surface expression was increased significantly by Q14703 , whereas P25116 remained unaffected . This response was associated with activation of the Akt , Erk , and p38 pathway and induction of P35354 but not P23219 . Q96RI0 -mediated induction of P35354 was prevented by the PI3K inhibitor LY ( 10 μM ) . Preincubation of human monocytes with Q14703 ( 1 μM ; 16 h ) resulted in an enhanced chemotaxis toward thrombin or to selective AP for Q96RI0 but not P25116 . Furthermore , down-regulation of Q96RI0 transcription with siRNA attenuated the chemotactic response to thrombin and AP4 . In conclusion , Q14703 enhances monocyte responses to thrombin via up-regulation of Q96RI0 expression , which promotes cell migration and P35354 abundance . This mechanism may facilitate monocyte recruitment to sites of vessel injury and inflammation . Functional consequences of Q14703 receptor modulation in rat oligodendroglial lineage cells . DB08868 ( FTY720 ) and its phosphorylated form FTY720P are modulators of sphingosine-1-phosphate ( Q14703 ) receptors , which are G-protein coupled receptors linked to cell migration and vascular maturation . The efficacy of FTY720 in autoimmune diseases such as multiple sclerosis and its animal models has been attributed to its inhibition of lymphocyte trafficking to target organs . In this study , we examined the role of Q14703 receptors in cultured rat oligodendrocytes ( OLGs ) and OLG progenitor cells ( OPCs ) using the active phosphorylated form of FTY720 . We found that ( 1 ) FTY720P improves the survival of neonatal rat OLGs during serum withdrawal , which is associated with the phosphorylation of extracellular signal regulated kinases ( P27361 /2 ) and Akt ; ( 2 ) FTY720P regulates OPC differentiation into OLGs in a concentration-dependent manner ; and ( 3 ) Q14703 receptors are differentially modulated by platelet-derived growth factor ( PDGF ) resulting in downregulation of Q9H228 and upregulation of P21453 in OPCs . In addition , siRNA studies revealed that P21453 participates in PDGF-induced OPC mitogenesis . We conclude that P21453 and Q9H228 serve different functions during oligodendroglial development , and possibly during remyelination . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . Prominence of central sphingosine-1-phosphate receptor-1 in attenuating aβ-induced injury by fingolimod . FTY720 ( fingolimod ) , the sphingosine-1-phosphate ( Q14703 ) analogue , has been experimentally indicated to exert substantial ameliorating effects in animal models of Alzheimer 's disease ( AD ) . The present work aims to answer whether central P21453 ( P21453 ) plays significant role in the impact of fingolimod in AD . To verify the prominence of central FTY720 phosphorylation , DMS ( sphingosine kinase inhibitor ) was infused intracerebrally in parallel with systemic FTY720 administration to prevent central formation of FTY720-P as the recognized active ligand for S1PRs . The corresponding P21453 modulation was also investigated using the pharmacological blockage of central P21453 by W123 . Both DMS and W123 were efficiently capable of suppressing FTY720-ameliorating effects in AD animals , either on memory deficit or on P36551 -II and P01375 -α expression . Our data conclude that experimental benefits of FTY720 in the context of AD depend on central P21453 modulation , as well as on Q14703 kinase activity in the brain . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . Sterol-regulatory-element-binding protein 2 and nuclear factor Y control human farnesyl diphosphate synthase expression and affect cell proliferation in hepatoblastoma cells . P14324 ( farnesyl diphosphate synthase ) catalyses the formation of farnesyl diphosphate , a key intermediate in the synthesis of cholesterol and isoprenylated cellular metabolites . P14324 is also the molecular target of nitrogen-containing bisphosphonates , which are used as bone-antiresorptive drugs in various disorders . In the present study , we characterized the sterol-response element and NF-Y ( nuclear factor Y ) -binding site in the human P14324 promoter . Using a luciferase assay , electrophoretic mobility-shift assay and chromatin immunoprecipitation assay , we demonstrated that these elements are responsible for the transcription of the P14324 gene , and that its transcriptional activation is mediated by Q12772 ( sterol-regulatory-element-binding protein 2 ) and NF-Y . We also investigated whether sterol-mediated P14324 expression is involved in the cell proliferation induced by zoledronic acid , an P14324 inhibitor . We show that the Q12772 - and NF-Y-mediated regulation of P14324 gene transcription modulates cell proliferation . These results suggest that Q12772 and NF-Y are required to trigger cell proliferation through the induction of P14324 expression and that the pharmacological action of zoledronic acid is involved in this pathway . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Regulation of extracellular signal-regulated kinase 1/2 influences hippocampal neuronal survival in a rat model of diabetic cerebral ischemia . Activation of extracellular signal-regulated kinase 1/2 has been demonstrated in acute brain ischemia . We hypothesized that activated extracellular signal-regulated kinase 1/2 can protect hippocampal neurons from injury in a diabetic model after cerebral ischemia/reperfusion . In this study , transient whole-brain ischemia was induced by four-vessel occlusion in normal and diabetic rats , and extracellular signal-regulated kinase 1/2 inhibitor ( U0126 ) was administered into diabetic rats 30 minutes before ischemia as a pretreatment . Results showed that the number of surviving neurons in the hippocampal P00915 region was reduced , extracellular signal-regulated kinase 1/2 phosphorylation and P12956 activity were decreased , and pro-apoptotic Bax expression was upregulated after intervention using U0126 . These findings demonstrate that inhibition of extracellular signal-regulated kinase 1/2 activity aggravated neuronal loss in the hippocampus in a diabetic rat after cerebral ischemia/reperfusion , further decreased DNA repairing ability and accelerated apoptosis in hippocampal neurons . P27361 /2 activation plays a neuroprotective role in hippocampal neurons in a diabetic rat after cerebral ischemia/reperfusion . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . DB03203 1-phosphate in amniotic fluid modulates cyclooxygenase-2 expression in human amnion-derived Q9NZQ3 cells . The metabolism of arachidonic acid , in particular the generation of prostaglandins ( PGs ) , has been proposed to play a key role in the regulation of labor . Moreover , several extracellular proteins have been reported to modulate PG synthesis in amnion cells . In this study , we found that lipid components dissolved in the amniotic fluid modulate PG synthesis in Q9NZQ3 human amnion cells and identified one of these components as a sphingosine 1-phosphate ( Q14703 ) . Q9NZQ3 cells express several Q14703 receptors including P21453 , O95136 , and Q99500 . When Q9NZQ3 cells were stimulated with Q14703 , DB00917 synthesis increased in a concentration-dependent manner , showing maximal activity at around 100 nM . Q14703 treatment also caused the up-regulation of cyclooxygenase-2 ( P35354 ) mRNA and protein , which was apparent within 3-12 h of stimulation . In terms of the intracellular signaling pathway of Q14703 -induced Q9NZQ3 cell activation , we found that Q14703 stimulated two kinds of MAPK , P29323 , and p38 kinase . We examined the roles of these two MAPKs in Q14703 -induced P35354 expression . Q14703 -induced P35354 expression was blocked completely by PD-98059 but not by SB-203580 , suggesting that P29323 has a critical role in the process . Transfection of P21453 or Q99500 but not of O95136 antisense oligonucleotide inhibited Q14703 -induced P35354 expression and DB00917 production in Q9NZQ3 cells , indicating the involvements of P21453 and Q99500 in the processes . This study demonstrates the physiological role of Q14703 in amniotic fluid and its effect on the modulation of P35354 expression and PGs synthesis in Q9NZQ3 cells . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Non-phosphorylated FTY720 induces apoptosis of human microglia by activating Q12772 . A synthetic analog of sphingosine named FTY720 ( DB08868 ) , phosphorylated by sphingosine kinase-2 , interacts with sphingosine-1-phosphate ( Q14703 ) receptors expressed on various cells . FTY720 suppresses the disease activity of multiple sclerosis ( MS ) chiefly by inhibiting Q14703 -dependent egress of autoreactive T lymphocytes from secondary lymphoid organs , and possibly by exerting anti-inflammatory and neuroprotective effects directly on brain cells . However , at present , biological effects of FTY720 on human microglia are largely unknown . We studied FTY720-mediated apoptosis of a human microglia cell line HMO6 . The exposure of HMO6 cells to non-phosphorylated FTY720 ( FTY720-non-P ) induced apoptosis in a dose-dependent manner with IC50 of 10.6 ± 2.0 μM , accompanied by the cleavage of caspase-7 and caspase-3 but not of caspase-9 . The apoptosis was inhibited by Z-DQMD-FMK , a caspase-3 inhibitor , but not by Pertussis toxin , a Gi protein inhibitor , suramin , a Q99500 / Q9H228 inhibitor , or W123 , a P21453 competitive antagonist , although HMO6 expressed P21453 , O95136 , and Q99500 . Furthermore , both phosphorylated FTY720 ( FTY720-P ) and SEW2871 , P21453 selective agonists , did not induce apoptosis of HMO6 . Genome-wide gene expression profiling and molecular network analysis indicated activation of transcriptional regulation by sterol regulatory element-binding protein ( SREBP ) in FTY720-non-P-treated HMO6 cells . Western blot verified activation of Q12772 in these cells , and apoptosis was enhanced by pretreatment with simvastatin , an activator of Q12772 , and by overexpression of the N-terminal fragment of Q12772 . These observations suggest that FTY720-non-P-induced apoptosis of HMO6 human microglia is independent of Q14703 receptor binding , and positively regulated by the Q12772 -dependent proapoptotic signaling pathway . Hydrogen peroxide induces P21453 receptors and sensitizes vascular endothelial cells to sphingosine 1-phosphate , a platelet-derived lipid mediator . DB03203 1-phosphate ( Q14703 ) is a platelet-derived angiogenic lipid growth factor , modulating G-protein-coupled Q14703 (1) receptors ( Q14703 (1)-R ) to activate endothelial nitric oxide synthase ( P29474 ) , as well as MAPK pathways in endothelial cells . We explored whether and how hydrogen peroxide ( H(2)O(2) ) , a representative reactive oxygen species , alters Q14703 (1)-R expression and influences Q14703 signaling in cultured bovine aortic endothelial cells ( BAECs ) . When BAECs are treated with pathophysiologically relevant concentrations of H(2)O(2) ( 150 microM for 30 min ) , Q14703 (1)-R protein expression levels are acutely augmented by approximately 30-fold in a dose-dependent fashion . When BAECs have been pretreated with H(2)O(2) , subsequent Q14703 stimulation ( 100 nM ) leads to a higher degree of P29474 enzyme activation ( assessed as intracellular cGMP content , 1.7 +/- 0.2-fold vs. no H(2)O(2) pretreatment groups , P < 0.05 ) , associated with a higher magnitude of phosphorylation responses of P29474 and MAPK P27361 /2 . Q99463 , an inhibitor of Src-family tyrosine kinase , abolished the effects of H(2)O(2) on both Q14703 (1)-R protein upregulation and enhanced BAEC responses to Q14703 . H(2)O(2) does not augment Q14703 (1) mRNA expression , whereas P15692 under identical cultures leads to increases in Q14703 (1) mRNA signals . Whereas H(2)O(2) attenuates proliferation of BAECs , addition of Q14703 restores growth responses of these cells . These results demonstrate that extracellularly administered H(2)O(2) increases Q14703 (1)-R expression and promotes endothelial responses for subsequent Q14703 treatment . These results may identify potentially important points of cross-talk between reactive oxygen species and sphingolipid pathways in vascular responses . Transduction of multiple effects of sphingosine 1-phosphate ( Q14703 ) on T cell functions by the P21453 G protein-coupled receptor . DB03203 1-phosphate ( Q14703 ) in blood , lymph , and immune tissues stimulates and regulates T cell migration through their Q14703 (1) ( endothelial differentiation gene encoded receptor-1 ) G protein-coupled receptors . We show now that Q14703 (1)Rs also mediate suppression of T cell proliferation and cytokine production . Uptake of [(3)H]thymidine by mouse P01730 T cells stimulated with anti-CD3 mAbs plus either anti- P10747 or P13232 was inhibited up to 50 % by 10(-9)-10(-6) M Q14703 . Suppression by Q14703 required Ca(2+) signaling and was reduced by intracellular DB02527 . Q14703 decreased P01730 T cell generation of P01579 and P05112 , without affecting P60568 . A Th1 line from D011.10 TCR transgenic mice without detectable Q14703 (1) was refractory to Q14703 until introduction of Q14703 (1) by retroviral transduction . Q14703 then evoked chemotaxis , inhibited chemotaxis to DB00833 -5 and DB00833 -21 , and suppressed Ag-stimulated proliferation and P01579 production . Thus , Q14703 (1) signals multiple immune functions of T cells as well as migration and tissue distribution . O95977 uses P04626 ( P04626 ) to regulate extracellular signal regulated kinase-1/2 in MDA-MB-453 breast cancer cells . We demonstrate here that the bioactive lipid sphingosine 1-phosphate ( Q14703 ) uses sphingosine 1-phosphate receptor 4 ( Q14703 (4) ) and human epidermal growth factor receptor 2 ( P04626 ) to stimulate the extracellular signal regulated protein kinase 1/2 ( P27361 /2 ) pathway in MDA-MB-453 cells . This was based on several lines of evidence . First , the Q14703 stimulation of P27361 /2 was abolished by JTE013 , which we show here is an Q14703 (2/4) antagonist and reduced by siRNA knockdown of Q14703 (4) . Second , the Q14703 -stimulated activation of P27361 /2 was almost completely abolished by a P04626 inhibitor ( ErbB2 inhibitor II ) and reduced by siRNA knockdown of P04626 expression . Third , phyto- Q14703 , which is an Q14703 (4) agonist , stimulated P27361 /2 activation in an Q14703 (4)- and P04626 -dependent manner . Fourth , FTY720 phosphate , which is an agonist at Q14703 (1,3,4,5) but not Q14703 (2) stimulated activation of P27361 /2 . Fifth , Q14703 stimulated the tyrosine phosphorylation of P04626 , which was reduced by JTE013 . P04626 which is an orphan receptor tyrosine kinase is the preferred dimerization partner of the P01133 receptor . However , P01133 -stimulated activation of P27361 /2 was not affected by siRNA knockdown of P04626 or by ErbB2 ( epidermal growth factor receptor 2 ( or P04626 ) ) inhibitor II in MDA-MB-453 cells . Moreover , Q14703 -stimulated activation of P27361 /2 does not require an P01133 receptor . Thus , Q14703 and P01133 function in a mutually exclusive manner . In conclusion , the magnitude of the signaling gain on the P27361 /2 pathway produced in response to Q14703 can be increased by P04626 in MDA-MB-453 cells . The linkage of Q14703 with an oncogene suggests that Q14703 and specifically Q14703 (4) may have an important role in breast cancer progression . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Farnesyl diphosphate synthase : the art of compromise between substrate selectivity and stereoselectivity . Farnesyl diphosphate ( FPP ) synthase catalyzes the consecutive head-to-tail condensations of isopentenyl diphosphate ( IPP , P01031 ) with dimethylallyl diphosphate ( DMAPP , P01031 ) and geranyl diphosphate ( GPP , Q99622 ) to give ( E,E ) -FPP ( C15 ) . The enzyme belongs to a genetically distinct family of chain elongation enzymes that install E-double bonds during each addition of a five-carbon isoprene unit . Analysis of the Q99622 and C15 products from incubations with avian P14324 reveals that small amounts of neryl diphosphate ( Z- Q99622 ) and ( Z,E ) -FPP are formed along with the E-isomers during the P01031 --> Q99622 and Q99622 --> C15 reactions . Similar results were obtained for P14324 from Escherichia coli , Artemisia tridentata ( sage brush ) , Pyrococcus furiosus , and Methanobacter thermautotrophicus and for GPP and FPP synthesized in vivo by E. coli P14324 . When ( R ) -[2-2H]IPP was a substrate for chain elongation , no deuterium was found in the chain elongation products . In contrast , the deuterium in ( S ) -[2-2H]IPP was incorporated into all of the products . Thus , the pro-R hydrogen at P06681 of IPP is lost when the E- and Z-double bond isomers are formed . The synthesis of Z-double bond isomers by P14324 during chain elongation is unexpected for a highly evolved enzyme and probably reflects a compromise between optimizing double bond stereoselectivity and the need to exclude DMAPP from the IPP binding site . Endogenous modulators and pharmacological inhibitors of histone deacetylases in cancer therapy . The class-I histone deacetylases ( HDACs ) Q13547 and Q92769 belong to a family of 11 zinc-dependent human HDACs and are overexpressed in many cancers . Inhibitors of these HDACs now in clinical trials show activity against several types of cancers . This review is focused on recent advances in both clinical and preclinical efforts to understand the basis for the actions of HDACis , with emphasis on implications for rational combinations with conventional or other targeted agents . We will address new perspectives on the molecular mechanisms by which HDACs act and how these actions relate to cancer . We will also review new evidence showing that HDACs are direct intracellular targets of the potent sphingolipid mediator Q14703 , the first identified endogenous nuclear regulator of these enzymes , linking sphingolipid metabolism in the nucleus to remodeling of chromatin and epigenetic regulation of gene expression . Understanding how endogenous molecules regulate HDAC activity in vivo may facilitate the search for safer and more effective anticancer drugs capable of interfering with HDAC functions in a highly specific manner . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Cyclical and dose-dependent responses of adult human mature oligodendrocytes to fingolimod . DB08868 is a sphingosine-1-phosphate ( Q14703 ) analogue that has been used in clinical trials as a systemic immunomodulatory therapy for multiple sclerosis . DB08868 readily accesses the central nervous system , raising the issue of its direct effects on neural cells . We assessed the effects of active fingolimod on dissociated cultures of mature , myelin-producing oligodendrocytes ( OLGs ) derived from adult human brain . Human OLGs express Q14703 receptor transcripts in relative abundance of Q9H228 > Q99500 > P21453 , with undetectable levels of O95977 . Low doses of fingolimod ( 100 pmol/L to 1 nmol/L ) induced initial membrane elaboration ( 2 days ) , subsequent retraction ( 4 days ) , and recurrence of extension with prolonged treatment ( 8 days ) . Higher doses ( 10 nmol/L to 1 mumol/L ) caused the opposite modulation of membrane dynamics . Retraction was rescued by co-treatment with the Q99500 / Q9H228 pathway antagonist , suramin , and was associated with RhoA-mediated cytoskeletal signaling . Membrane elaboration was mimicked using the P21453 agonist SEW2871 . DB08868 rescued human OLGs from serum and glucose deprivation-induced apoptosis , which was reversed with suramin co-treatment and mimicked using an Q9H228 agonist . High doses of fingolimod induced an initial down-regulation of Q9H228 mRNA levels relative to control ( 4 hours ) , subsequent up-regulation ( 2 days ) , and recurrent down-regulation ( 8 days ) . P21453 mRNA levels were inversely regulated compared with Q9H228 . These results indicate that fingolimod modulates maturity- and species-specific OLG membrane dynamics and survival responses that are directly relevant for myelin integrity . P05231 and P10145 release is mediated via multiple signaling pathways after stimulating dendritic cells with lysophospholipids . Lysophosphatidic acid ( P08519 ) and sphingosine 1-phosphate ( Q14703 ) are bioactive lipid mediators , which are known to play major roles in allergic reactions as well as in tumor pathogenesis . Here , the biological activities and signal pathways of these lysophospholipids ( LPLs ) in dendritic cells ( DCs ) were characterized further . Flow cytometric and immunoblot analyses indicate that immature as well as mature DCs express the P06858 receptors P21453 , Q99500 , Q9H228 , and Q9HBW0 , but not O95136 , O95977 , Q92633 , or Q9UBY5 . Moreover , enzyme-linked immunosorbent assay experiments demonstrate that simultaneous addition of these LPLs to immature DCs in the presence of lipopolysaccharide enhanced the secretion of the inflammatory cytokines interleukin ( IL ) -6 and P10145 in maturing DCs . In contrast , no modification of P05231 or P10145 release was observed after exposure of mature DCs to LPLs alone . In addition , studies with pertussis toxin and mitogen-activated protein kinase ( MAPK ) kinase inhibitor PD98059 suggested that Gi proteins and MAPK pathway are involved in these P06858 -induced cell responses . Corroborating these findings , we observed that LPLs induce the phosphorylation of extracellular signal-regulated kinase 1/2 in immature DCs but not in mature DCs . Further analyses show that inhibitors of phosholipase D , Rho , and protein kinase C also inhibited the P06858 -induced release of P05231 and P10145 . Therefore , our findings suggest that lipopolysaccharide in DCs uncouples P06858 receptors from the signal-transducing machinery during maturation and that exposure of LPLs at early time-points to maturing DCs modifies the proinflammatory capacity of mature DCs . DB11320 contributes to tissue remodeling via periostin expression . DB11320 is thought to have a critical role in the synthesis of extracellular matrix in skin and may be involved in tissue remodeling of allergic diseases . Recent studies revealed that periostin , a matricelluar protein , contributed to tissue remodeling ; however , a link between periostin and histamine remains unproven . We investigated whether periostin was involved in histamine-induced collagen production . Cultured dermal fibroblasts derived from wild-type ( WT ) or periostin knockout ( PN(-/-) ) mice were stimulated with histamine , and then collagen and periostin production was evaluated . DB11320 induced collagen gene expression in WT fibroblasts in the late phase but not in the early phase , whereas no effect on collagen expression was observed in histamine-stimulated PN(-/-) fibroblasts . In WT fibroblasts , histamine directly induced periostin expression in a dose-dependent manner , and an H1 receptor antagonist blocked both periostin and collagen expression . DB11320 activated extracellular signal-regulated kinase 1/2 ( P27361 /2 ) through the H1 receptor . Q15063 induction was inhibited by either H1 antagonist or P27361 /2 inhibitor treatment in vitro and was attenuated in P35367 (-/-) mice . Elevated expression of periostin was found in lesional skin from atopic dermatitis patients . These results suggest that histamine mediates periostin induction and collagen production through activation of the H1 receptor-mediated P27361 /2 pathway ; furthermore , histamine may accelerate the chronicity of atopic dermatitis . P21453 as a useful target for treatment of multiple sclerosis . DB03203 1-phosphate ( Q14703 ) , a lysophospholipid mediator , is generated from sphingosine by sphingosine kinases and binds five known cell surface receptors . P21453 ( P21453 ) plays an essential role in lymphocyte egress from secondary lymphoid organs ( Q12791 ) , as evinced by the inability of lymphocytes to exit from the Q12791 in mice lacking lymphocytic P21453 . DB08868 hydrochloride ( FTY720 ) is a first-in-class , orally active , Q14703 receptor modulator with a structure closely related to sphingosine . FTY720 was first synthesized by chemical modification of a natural product , myriocin . FTY720 is effectively converted to an active metabolite , FTY720 phosphate ( FTY720-P ) by sphingosine kinases . FTY720-P shows high affinity to 4 of the Q14703 receptors ( P21453 , Q99500 , O95977 , and Q9H228 ) . In particular , FTY720-P strongly induces internalization and degradation of P21453 , inhibits Q14703 responsiveness of lymphocytes in the Q12791 , and acts as a functional antagonist at lymphocytic P21453 . Consequently , FTY720 inhibits P21453 -dependent lymphocyte egress from the Q12791 to decrease circulation of lymphocytes including autoreactive Th17 cells and is highly effective in experimental autoimmune encephalomyelitis ( EAE ) , an animal model of multiple sclerosis ( MS ) . Because FTY720 shows a superior efficacy in relapsing remitting MS patients compared to intramuscular interferon-β-1a ( Avonex® ) , P21453 is presumed to be a useful target for the therapy of MS . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Q9NYA1 mediates head & neck squamous cell carcinoma invasion through sphingosine 1-phosphate receptor 1 . BACKGROUND : Head and neck squamous cell carcinoma ( HNSCC ) is characterized by aggressive loco-regional invasion . DB03203 kinase1 ( SphK1 ) , an enzyme in sphingolipid metabolism , is emerging as a key player in HNSCC pathology . The observation that SphK1 is overexpressed in all HNSCC stages and is associated with depth of tumor invasion , metastasis and clinical failure underscores the importance of SphK1 in HNSCC pathology . Still , the mechanisms underlying SphK1 regulation of invasion have not been delineated . Therefore , we sought to mechanistically describe how SphK1 regulates invasion in HNSCC . METHODS : Invasion assays were used to measure invasive ability of SphK1 overexpressing human tongue squamous cell carcinoma ( SCC-25 cells ) . Western blotting , quantitative qPCR , ELISA and zymography were used to measure the effect of SphK1 and sphingosine 1-phoshate receptor 1 ( P21453 ) on invasion measures , P08253 /9 , P12830 , P00533 , P05231 / P40763 , in SCC-25 cells . RESULTS : SphK1 expression is elevated in cells with an invasive phenotype as compared to non-invasive phenotype . We show SphK1 overexpression increased P01133 -induced P00533 / P29323 and AKT activity , increased matrix metalloproteinase ( MMP ) -2/9 mRNA and reduced P12830 . SphK1 overexpression also increased P05231 concentration and P01133 -induced P40763 phosphorylation , exemplifying that SphK1 modulates P05231 / P40763 signaling . Notably , we show that P21453 knockdown reduced P05231 / P40763 signaling , representing another pathway by which SphK1/ Q14703 regulates invasion . CONCLUSIONS : Taken together , our data suggest that SphK1 sits at the hub of multiple key signaling cascades , all which have been implicated in the regulation of invasiveness , making SphK1 an attractive target for the development of HNSCC therapies . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . DB03203 1-phosphate rescues canine LPS-induced acute lung injury and alters systemic inflammatory cytokine production in vivo . Q14703 has been demonstrated to protect against the formation of lipopolysaccharide ( LPS ) -induced lung edema when administered concomitantly with LPS . In the current study , we sought to determine the effectiveness of Q14703 to attenuate lung injury in a translationally relevant canine model of ALI when administered as rescue therapy . Secondarily , we examined whether the attenuation of LPS-induced physiologic lung injury after administration of Q14703 was , at least in part , caused by an alteration in local and/or systemic inflammatory cytokine expression . We examined 18 , 1-year-old male beagles prospectively in which we instilled bacterial LPS ( 2-4 mg/kg ) intratracheally followed in 1 h with intravenous Q14703 ( 85 microg/kg ) or vehicle and 8 h of high-tidal-volume mechanical ventilation . Q14703 attenuated the formation of Q(s)/Q(t) ( 32 % ) , and both the presence of protein ( 72 % ) and neutrophils ( 95 % ) in BAL fluid compared with vehicle controls . Although lung tissue inflammatory cytokine production was found to vary regionally throughout the LPS-injured lung , Q14703 did not alter the expression pattern . Similarly , BAL cytokine production was not altered significantly by intravenous Q14703 in this model . Interestingly , Q14703 potentiated the LPS-induced systemic production of 3 inflammatory cytokines , P01375 ( 6-fold ) , KC ( 1.2-fold ) , and P05231 ( 3-fold ) , without resulting in end-organ dysfunction . In conclusion , intravenous Q14703 reduces inflammatory lung injury when administered as rescue therapy in our canine model of LPS-induced ALI . This improvement is observed in the absence of changes in local pulmonary inflammatory cytokine production and an augmentation of systemic inflammation . DB08868 ( FTY720 ) enhances remyelination following demyelination of organotypic cerebellar slices . Remyelination , which occurs subsequent to demyelination , contributes to functional recovery and is mediated by oligodendrocyte progenitor cells ( OPCs ) that have differentiated into myelinating cells . Therapeutics that impact remyelination in the CNS could be critical determinants of long-term functional outcome in multiple sclerosis ( MS ) . DB08868 is a Q14703 receptor modulator in MS clinical trials due to systemic anti-inflammatory properties , yet may impact cells within the CNS by crossing the blood-brain barrier . Previous studies using isolated dissociated cultures indicate that neural cells express Q14703 receptors and respond to receptor engagement . Our objective was to assess the effects of fingolimod on myelin-related processes within a multicellular environment that maintains physiological cell-cell interactions , using organotypic cerebellar slice cultures . DB08868 treatment had no impact on myelin under basal conditions . DB08868 treatment subsequent to lysolecithin-induced demyelination enhanced remyelination and process extension by OPCs and mature oligodendrocytes , while increasing microglia numbers and immunoreactivity for the astrocytic marker glial fibrillary acidic protein . The number of phagocytosing microglia was not increased by fingolimod . Using Q14703 receptor specific agonists and antagonists , we determined that fingolimod-induced effects on remyelination and astrogliosis were mediated primarily through Q99500 and Q9H228 , whereas enhanced microgliosis was mediated through P21453 and Q9H228 . Taken together , these data demonstrate that fingolimod modulates multiple neuroglial cell responses , resulting in enhanced remyelination in organotypic slice cultures that maintain the complex cellular interactions of the mammalian brain . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Chronic inhibition of farnesyl pyrophosphate synthase attenuates cardiac hypertrophy and fibrosis in spontaneously hypertensive rats . P14324 ( FPPS ) , an essential enzyme in the mevalonate pathway , was reported to be upregulated in young spontaneously hypertensive rats ( SHR ) when compared with Wistar-Kyoto ( WKY ) rats , and this was accompanied by development of left ventricular hypertrophy . Five-week-old rats were daily gavaged with vehicle or an FPPS inhibitor ( alendronate , 1 or 10 mg/kg ) and blood pressures was monitored by the tail-cuff method every other week . Twelve weeks of alendronate treatment attenuated the left ventricular weight to body weight ratio ( LVW/BW ) , hydroxyproline content , collagen deposition in the interstitia , and gene expression of atrial natriuretic peptide , B-type natriuretic peptide , and procollagen type I/III in the SHR left ventricle , all of which were significantly higher in SHRs than in WKY rats . Furthermore , long-term treatment with an FPPS inhibitor significantly reduced RhoA activation , P29323 phosphorylation , and TGF-beta1 expression in the SHR left ventricle , all of which were upregulated more in SHRs than in WKY rats . In conclusion , chronic treatment with an FPPS inhibitor attenuates the development of cardiac hypertrophy and fibrosis , and the suppression of P27361 /2 phosphorylation and TGF-beta1 expression with inhibition of RhoA activation may be an important mechanism . DB03203 -1-phosphate receptors regulate individual cell behaviours underlying the directed migration of prechordal plate progenitor cells during zebrafish gastrulation . During vertebrate gastrulation , cells forming the prechordal plate undergo directed migration as a cohesive cluster . Recent studies revealed that P12830 -mediated coherence between these cells plays an important role in effective anterior migration , and that platelet-derived growth factor ( Pdgf ) appears to act as a guidance cue in this process . However , the mechanisms underlying this process at the individual cell level remain poorly understood . We have identified miles apart ( mil ) as a suppressor of defective anterior migration of the prospective prechordal plate in silberblick ( slb ) /wnt11 mutant embryos , in which P12830 -mediated coherence of cell movement is reduced. mil encodes Edg5 , a sphingosine-1-phosphate ( Q14703 ) receptor belonging to a family of five G-protein-coupled receptors ( S1PRs ) . Q14703 is a lipid signalling molecule that has been implicated in regulating cytoskeletal rearrangements , cell motility and cell adhesion in a variety of cell types . We examined the roles of Mil in anterior migration of prechordal plate progenitor cells and found that , in slb embryos injected with mil-MO , cells migrate with increased motility but decreased directionality , without restoring the coherence of cell migration . This indicates that prechordal plate progenitor cells can migrate effectively as individuals , as well as in a coherent cluster of cells . Moreover , we demonstrate that Mil regulates cell motility and polarisation through Pdgf and its intracellular effecter PI3K , but modulates cell coherence independently of the Pdgf/PI3K pathway , thus co-ordinating cell motility and coherence . These results suggest that the net migration of prechordal plate progenitors is determined by different parameters , including motility , persistence and coherence . DB08868 : direct CNS effects of sphingosine 1-phosphate ( Q14703 ) receptor modulation and implications in multiple sclerosis therapy . DB08868 is the first oral disease-modifying therapy approved for relapsing forms of multiple sclerosis ( MS ) . Following phosphorylation in vivo , the active agent , fingolimod phosphate ( fingolimod-P ) , acts as a sphingosine 1-phosphate ( Q14703 ) receptor modulator , binding with high affinity to four of the five known Q14703 receptors ( P21453 , Q99500 , O95977 and Q9H228 ) . The mechanism of action of fingolimod in MS has primarily been considered as immunomodulatory , whereby fingolimod-P modulates P21453 on lymphocytes , selectively retaining autoreactive lymphocytes in lymph nodes to reduce damaging infiltration into the central nervous system ( CNS ) . However , emerging evidence indicates that fingolimod has direct effects in the CNS in MS . For example , in the MS animal model of experimental autoimmune encephalomyelitis ( EAE ) , fingolimod is highly efficacious in both a prophylactic and therapeutic setting , yet becomes ineffective in animals selectively deficient for P21453 on astrocytes , despite maintained normal immunologic receptor expression and functions , and Q14703 -mediated immune activities . Here we review Q14703 signaling effects relevant to MS in neural cell types expressing Q14703 receptors , including astrocytes , oligodendrocytes , neurons , microglia and dendritic cells . The direct effects of fingolimod on these CNS cells observed in preclinical studies are discussed in view of the functional consequences of reducing neurodegenerative processes and promoting myelin preservation and repair . The therapeutic implications of Q14703 modulation in the CNS are considered in terms of the clinical outcomes of MS , such as reducing MS-related brain atrophy , and other CNS disorders . Additionally , we briefly outline other existing and investigational MS therapies that may also have effects in the CNS . Synthesis and evaluation of fluorinated fingolimod ( FTY720 ) analogues for sphingosine-1-phosphate receptor molecular imaging by positron emission tomography . DB03203 -1-phosphate ( Q14703 ) is a lysophospholipid that evokes a variety of biological responses via stimulation of a set of cognate G-protein coupled receptors ( GPCRs ) : P21453 - Q9H228 . Q14703 and its receptors ( S1PRs ) play important roles in the immune , cardiovascular , and central nervous systems and have also been implicated in carcinogenesis . Recently , the Q14703 analogue DB08868 ( FTY720 ) has been approved for the treatment of patients with relapsing multiple sclerosis . This work presents the synthesis of various fluorinated structural analogues of FTY720 , their in vitro and in vivo biological testing , and their development and application as [(18)F]radiotracers for the study of S1PR biodistribution and imaging in mice using small-animal positron emission tomography ( PET ) . DB03203 1-phosphate signalling in cancer . There is an increasing body of evidence demonstrating a critical role for the bioactive lipid Q14703 ( sphingosine 1-phosphate ) in cancer . Q14703 is synthesized and metabolized by a number of enzymes , including sphingosine kinase , Q14703 lyase and Q14703 phosphatases . Q14703 binds to cell-surface G-protein-coupled receptors ( P21453 - Q9H228 ) to elicit cell responses and can also regulate , by direct binding , a number of intracellular targets such as HDAC ( histone deacetylase ) 1/2 to induce epigenetic regulation . Q14703 is involved in cancer progression including cell transformation/oncogenesis , cell survival/apoptosis , cell migration/metastasis and tumour microenvironment neovascularization . In the present paper , we describe our research findings regarding the correlation of sphingosine kinase 1 and Q14703 receptor expression in tumours with clinical outcome and we define some of the molecular mechanisms underlying the involvement of sphingosine kinase 1 and Q14703 receptors in the formation of a cancer cell migratory phenotype . The role of sphingosine kinase 1 in the acquisition of chemotherapeutic resistance and the interaction of Q14703 receptors with oncogenes such as P04626 is also reviewed . We also discuss novel aspects of the use of small-molecule inhibitors of sphingosine kinase 1 in terms of allosterism , ubiquitin-proteasomal degradation of sphingosine kinase 1 and anticancer activity . Finally , we describe how Q14703 receptor-modulating agents abrogate Q14703 receptor-receptor tyrosine kinase interactions , with potential to inhibit growth-factor-dependent cancer progression . P35354 induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways . The functional significance of protease-activated receptors ( PARs ) in endothelial cells is largely undefined , and the intracellular consequences of their activation are poorly understood . Here , we show that the serine protease thrombin , a P25116 -selective peptide ( TFLLRN ) , and SLIGKV ( P55085 -selective peptide ) induce cyclooxygenase-2 ( P35354 ) protein and mRNA expression in human endothelial cells without modifying P23219 expression . P35354 induction was accompanied by sustained production of 6-keto-PGF1alpha , the stable hydrolysis product of prostacyclin , and this was inhibited by indomethacin and the P35354 -selective inhibitor NS398 . P25116 and P55085 stimulation rapidly activated both P27361 /2 and p38MAPK , and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha formation . Thrombin and peptide agonists of P25116 and P55085 increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus , and this , as well as PAR-induced 6-keto-PGF1alpha synthesis , was inhibited by co-infection with adenovirus encoding wild-type or mutated ( Y42F ) P25963 . Thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG-132 . Activation of P25116 or P55085 promoted nuclear translocation and phosphorylation of p65-NF-kappaB , and thrombin-induced but not P55085 -induced p65-NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK . Activation of Q96RI0 by AYPGKF increased phosphorylation of P27361 /2 and p38MAPK without modifying NF-kappaB activation or P35354 induction . Our data show that P25116 and P55085 , but not Q96RI0 , are coupled with P35354 expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on P27361 /2 , p38MAPK , and P25963 -dependent NF-kappaB activation . Activation of P00533 promotes squamous carcinoma SCC10A cell migration and invasion via inducing EMT-like phenotype change and P14780 -mediated degradation of P12830 . P00533 is a potent stimulator of invasion and metastasis in head and neck squamous cell carcinomas ( HNSCC ) . However , the mechanism by which P00533 may stimulate tumor cell invasion and metastasis still need to be elucidated . In this study , we showed that activation of P00533 by P01133 in HNSCC cell line SCC10A enhanced cell migration and invasion , and induced loss of epitheloid phenotype in parallel with downregulation of P12830 and upregulation of P19022 and vimentin , indicating that P00533 promoted SCC10A cell migration and invasion possibly by an epithelial to mesenchymal transition ( EMT ) -like phenotype change . Interestingly , activation of P00533 by P01133 induced production of matrix metalloproteinase-9 ( P14780 ) and soluble P12830 ( sE-cad ) , and knockdown of P14780 by siRNA inhibited sE-cad production induced by P01133 in SCC10A . Moreover , both P14780 knockdown and P12830 overexpression inhibited cell migration and invasion induced by P01133 in SCC10A . The results indicate that P00533 activation promoted cell migration and invasion through inducing P14780 -mediated degradation of P12830 into sE-cad . Pharmacologic inhibition of P00533 , MEK , and PI3K kinase activity in SCC10A reduced phosphorylated levels of P27361 /2 and AKT , production of P14780 and sE-cad , cell migration and invasion , and expressional changes of EMT markers ( P12830 and P19022 ) induced by P01133 , indicating that P00533 activation promotes cell migration and invasion via P27361 /2 and PI3K-regulated P14780 / P12830 signaling pathways . Taken together , the data suggest that P00533 activation promotes HNSCC SCC10A cell migration and invasion by inducing EMT-like phenotype change and P14780 -mediated degradation of P12830 into sE-cad related to activation of P27361 /2 and PI3K signaling pathways . Early changes in DB09150 incorporation by breast cancer cells treated with trastuzumab in normoxic conditions : role of the Akt-pathway , glucose transport and HIF-1α . HER-2 overexpression does not guarantee response to P04626 -targeting drugs such as trastuzumab , which is cardiotoxic and expensive , so early detection of response status is crucial . Factors influencing [(18)F] DB09150 incorporation in the timeframe of cell signalling down-regulation subsequent to trastuzumab treatment are investigated to provide a better understanding of the relationship between growth response and modulation of [(18)F] DB09150 incorporation . HER-2-overexpressing breast tumour cell lines , MDA-MB-453 , SKBr3 and BT474 and MDA-MB-468 ( P04626 non-over-expressor ) were treated with trastuzumab ( 4 h ) and probed for AKT , pAKT , P27361 /2 , pERK1/2 and HIF-1α to determine early signalling pathway inhibitory effects of trastuzumab . Cells incubated with trastuzumab and/or PI3K inhibitor LY294002 and P27361 /2 inhibitor U0126 and glucose transport and [(18)F] DB09150 incorporation measured . Cell lines expressed AKT , pAKT , P27361 /2 and pERK1/2 but not HIF-1α . DB00072 treatment decreased pAkt but not pERK1/2 levels . DB00072 did not further inhibit AKT when maximally inhibited with LY294002 . Treatment with LY294002 and trastuzumab for 4 h decreased [(18)F] DB09150 incorporation in BT474 and MDA-MB-453 but not SKBr3 cells . LY294002 inhibited glucose transport by each cell line , but the glucose transport rate was tenfold higher by SKBr3 cells than BT474 and MDA-MB-453 cells . AKT-induced uptake of [(18)F] DB09150 was found to be HIF-1α independent in breast cancer cell lines . AKT inhibition level and tumour cell glucose transport rate can influence whether or not PI3K inhibitors affect [(18)F] DB09150 incorporation which may account for the variation in preclinical and clinical findings associated with [(18)F] DB09150 -PET in response to trastuzumab and other HER-2 targeting drugs . Distinct energy requirements for human memory P01730 T-cell homeostatic functions . Differentiation and activation of P01730 memory T cells ( T(mem) cells ) require energy from different sources , but little is known about energy sources for maintenance and surveillance activities of unactivated T(mem) cells . Mitochondrial fatty acid oxidation ( FAO ) in human unactivated P01730 T(mem) cells was significantly enhanced by inhibition of glycolysis , with respective means of 1.7- and 4.5-fold for subjects < 45 yr and > 65 yr , and by stimulation of AMP-activated protein kinase , with respective means of 1.3- and 5.2-fold . However , Q99731 and sphingosine 1-phosphate ( Q14703 ) , which control homeostatic lymphoid trafficking of unactivated T(mem) cells , altered FAO and glycolysis only minimally or not at all . Inhibition of P01730 T(mem)-cell basal FAO , but not basal glycolysis , significantly suppressed Q99731 - and Q14703 -mediated adherence to collagen by > 50 and 20 % , respectively , and chemotaxis by > 20 and 50 % . Apoptosis of unactivated T(mem) cells induced by P60568 deprivation or Q99731 was increased significantly by > 150 and 70 % , respectively , with inhibition of FAO and by > 110 and 30 % with inhibition of glycolysis . Anti-TCR antibody activation of T(mem) cells increased their chemotaxis to P13501 , which was dependent predominantly on glycolysis rather than FAO . The sources supplying energy for diverse functions of unactivated T(mem) cells differ from that required for function after immune activation . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Hypoxic/normoxic preconditioning increases endothelial differentiation potential of human bone marrow CD133+ cells . CD133+ cells are hemangioblasts that have capacity to generate into both hematopoietic and endothelial cells ( ECs ) . Hypoxia/normoxia has shown to be the regulator of the balance between stemness and differentiation . In this study we performed Agilent 's whole human genome oligo microarray analysis and examined the differentiation potential of the bone-marrow-derived CD133+ cells after hypoxic/normoxic preconditioning of CD133+ cells . Results showed that there was no significant increase in erythroid colony forming unit ( CFU-E ) and CFU-granulocyte , erythrocyte , monocyte , and megakaryocyte formation with cells treated under hypoxia/normoxia . However , a significant increment of EC forming unit at 24 h ( 143.2 +/- 8.0 % ) compared to 0 h ( 100 +/- 11.4 % ) was observed in CFU-EC analysis . Reverse transcription-polymerase chain reaction and immunostaining analysis showed that the differentiated cells diminished hematopoietic stem cell surface markers and acquired the gene markers and functional phenotype of ECs . The transcriptome profile revealed a cluster of 232 downregulated and 498 upregulated genes in cells treated for 24 h under hypoxia . The upregulated genes include angiogenic genes , angiogenic growth factor genes , angiogenic cytokine and chemokine genes , as well as angiogenic-positive regulatory genes , including Q14512 , PDGFB , Q16663 , P48061 , P80162 , P05231 , P21246 , O14944 , P04626 , O95136 , P11487 , Q92913 , Q99988 , P05412 , L1CAM , Q02297 , P08138 , and PDGFB . On the other hand , angiogenesis inhibitors and related genes , including P29459 , P98177 , Q9NY15 , and P16035 , are downregulated . Taken together , hypoxic/normoxic preconditioning may lead to the differentiation of CD133+ cells toward endothelial lineage , which may improve the current clinical trial studies . DB03203 1-phosphate mediates proliferation and survival of mesoangioblasts . Mesoangioblasts are stem cells capable of differentiating in various mesodermal tissues and are presently regarded as suitable candidates for cell therapy of muscle degenerative diseases , as well as myocardial infarction . The enhancement of their proliferation and survival after injection in vivo could greatly improve their ability to repopulate damaged tissues . In this study , we show that the bioactive sphingolipid sphingosine 1-phosphate ( Q14703 ) regulates critical functions of mesoangioblast cell biology . Q14703 evoked a full mitogenic response in mesoangioblasts , measured by labeled thymidine incorporation and cell counting . Moreover , Q14703 strongly counteracted the apoptotic process triggered by stimuli as diverse as serum deprivation , P06681 -ceramide treatment , or staurosporine treatment , as assessed by cell counting , as well as histone-associated fragments and caspase-3 activity determinations . Q14703 acts both as an intracellular messenger and through specific membrane receptors . Real-time polymerase chain reaction analysis revealed that mesoangioblasts express the Q14703 -specific receptor Q99500 and , to a minor extent , P21453 and O95136 . By using Q14703 receptor subtype-specific agonists and antagonists , we found that the proliferative response to Q14703 was mediated mainly by O95136 . By contrast , the antiapoptotic effect did not implicate Q14703 receptors . These findings demonstrate an important role of Q14703 in mesoangioblast proliferation and survival and indicate that targeting modulation of Q14703 -dependent signaling pathways may be used to improve the efficiency of muscle repair by these cells . Disclosure of potential conflicts of interest is found at the end of this article . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Novel cinnamyl hydroxyamides and 2-aminoanilides as histone deacetylase inhibitors : apoptotic induction and cytodifferentiation activity . Four novel series of cinnamyl-containing histone deacetylase ( HDAC ) inhibitors 1-4 are described , containing hydroxamate ( 1 and 3 ) or 2-aminoanilide ( 2 and 4 ) derivatives . When screened against class I ( maize HD1-B and human Q13547 ) and class II ( maize HD1-A and human P56524 ) HDACs , most hydroxamates and 2-aminoanilides displayed potent and selective inhibition toward class I enzymes . Immunoblotting analyses performed in U937 leukemia cells generally revealed high acetyl-H3 and low acetyl-α-tubulin levels . Exceptions are compounds 3 f-i , 3 m-o , and 4 k , which showed higher tubulin acetylation than DB02546 . In U937 cells , cell-cycle blockade in either the G₂/M or G₁/S phase was observed with 1-4 . Five hydroxamates ( compounds 1 h-l ) effected a two- to greater than threefold greater percent apoptosis than DB02546 , and in the CD11c cytodifferentiation test some 2-aminoanilides belonging to both series 2 and 4 were more active than MS-275 . The highest-scoring derivatives in terms of apoptosis ( 1 k , 1 l ) or cytodifferentiation ( 2 c , 4 n ) also showed antiproliferative activity in U937 cells , thus representing valuable tools for study in other cancer contexts .

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Nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase isoforms during neuronal apoptosis . Treatment with cytosine beta-D-arabinoside ( AraC ; 300 microM ) induced a time-dependent accumulation of glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) protein in nuclei purified from cultured cerebellar granule cells , with a concomitant degradation of lamin B1 , a nuclear membrane protein and a substrate of P42574 /caspase-3 . Moreover , DB00128 - DB00142 - DB00161 - DB00128 -fluoromethyl ketone ( DEVD-fmk ) , a P42574 -selective antagonist , dose-dependently suppressed AraC-induced apoptosis of these neurons . Nuclear accumulation of P04406 protein was associated with a progressive decrease in the activity of uracil-DNA glycosylase ( P13051 ) , one of the nuclear functions of P04406 . The nuclear dehydrogenase activity of P04406 was initially increased after treatment and then decreased parallel to P13051 activity . Six P04406 isoforms were detected in the nuclei of AraC-treated cells . The more alkaline isoforms , 1-3 , constituted the bulk of the nuclear P04406 , and the remaining isoforms , 4-6 , were the minor species . Levels of all six isoforms were increased after treatment with AraC for 16 h ; a 4-h treatment increased levels of only isoforms 4 and 5 . Thus , it appears that various P04406 isoforms are differentially regulated and may have distinct apoptotic roles . Pretreatment with P04406 antisense oligonucleotide blocked the nuclear translocation of P04406 isoforms , and the latter process occurred concurrently with a decrease in cytosolic P04406 isoforms . Sodium nitroprusside-induced NAD labeling of nuclear P04406 showed a 60 % loss of P04406 labeling after AraC treatment , suggesting that the active site of P04406 may be covalently modified , denatured , or improperly folded . The unfolded protein response elicited by denatured P04406 may contribute to AraC-induced neuronal death . Single-prolonged stress induce changes of P62158 /CaMKIIα in the rats of dorsal raphe nucleus . Ca2+/calmodulin-dependent protein kinase IIα ( CaMKIIα ) is identified as a Ca2+-dependent kinase in brain involved in the activation of DB00150 hydroxylase ( P17752 ) acting through direct phosphorylation of P17752 , and playing key roles in the signaling pathways initiated by various G protein-coupled 5-HT receptors . The goal of this study is to detect whether there are changes of P62158 and CaMKIIα in dorsal raphe nucleus in the rats exposed to single-prolonged stress ( P49903 ) , which is a model employed in post-traumatic stress disorder ( PTSD ) study extensively . A total of 90 male Wistar rats were randomly divided into a normal control group and P49903 groups of 7d , 14d . The changes of P62158 /CaMKIIα were detected by immunohistochemistry , reverse transcription-polymerase chain reaction and western blot . Our results demonstrate that both expressions of P62158 and CaMKIIα significantly increase ( P < 0.001 ) in the P49903 7d group than that in the control group , and then decreased dramatically ( P < 0.001 ) 14 days after P49903 . Our results confirm that P49903 induce changes of P62158 /CaMKIIα in the dorsal raphe nucleus . Changes of P62158 /CaMKIIα may be associated with the activation of P08908 receptor , and may contribute to the progress of molecular mechanism of PTSD . Discriminative stimulus effects of gamma-hydroxybutyrate ( DB01440 ) and its metabolic precursor , DB04699 ( Q9BVC4 ) in rats . RATIONALE : Gamma-hydroxybutyrate ( DB01440 ) is becoming an increasingly popular drug of abuse . Metabolic precursors of DB01440 , DB04699 ( Q9BVC4 ) and 1,4-butanediol ( BDL ) , are commercially available industrial solvents that may also present potential health risks . Relatively little is known about the neurobehavioral effects of DB01440 and its precursors . OBJECTIVE : The aim of the present investigation was to characterize the discriminative stimulus effects of DB01440 and its precursor , Q9BVC4 . METHODS : Male Sprague-Dawley rats were trained to discriminate DB01440 [ 300 mg/kg , i.g. ; n=16 ] or Q9BVC4 ( 150 mg/kg , i.p. ; n=8 ) from vehicle under a fixed ratio 20 ( FR 20 ) schedule of food reinforcement . Stimulus generalization tests were then conducted with several compounds . RESULTS : DB01440 and Q9BVC4 produced cross-generalization and BDL was fully substituted for both DB01440 and Q9BVC4 . Two benzodiazepines , alprazolam and diazepam , and the P08908 agonist , buspirone , did not substitute for either training drug nor did ethanol or the DB01221 antagonists , PCP and ketamine . The DB01440 antagonist , NCS-382 , and the GABA(B) antagonist , CGP-35348 , blocked the discriminative stimulus effects of DB01440 but not those of Q9BVC4 . CONCLUSIONS : These findings suggest that DB01440 and its metabolic precursors produce similar subjective effects that differ from those of other sedative-hypnotic drugs . Further investigations into the neurochemical actions underlying the subjective effects of these drugs are warranted . Fibrinolytic activity in the inner ear of human beings and guinea pigs . Fibrinolytic activity in the inner ear of human beings and guinea pigs was assayed qualitatively and quantitatively . It was found that fibrinolytic activity of the stria vascularis was caused by a conversion of plasminogen to plasmin by tissue plasminogen activator and was moderate in degree compared with that of other organs in both species . In human beings , endolymph showed higher plasminogen activator activity than that of P04141 . P00747 activator activity was not detected in either endolymph or perilymph of guinea pigs . Thrombin infusion diminished plasminogen activator activity in the stria vascularis of guinea pigs . In vitro prostanoid release from spinal cord following peripheral inflammation : effects of DB05875 , DB01221 and capsaicin . 1. Spinal prostanoids are implicated in the development of thermal hyperalgesia after peripheral injury , but the specific prostanoid species that are involved are presently unknown . The current study used an in vitro spinal superfusion model to investigate the effect of DB05875 ( SP ) , N-methyl-d-aspartate ( DB01221 ) , and capsaicin on multiple prostanoid release from dorsal spinal cord of naive rats as well as rats that underwent peripheral injury and inflammation ( knee joint kaolin/carrageenan ) . 2 . In naive rat spinal cords , DB00917 and 6-keto-PGF1alpha , but not TxB2 , levels were increased after inclusion of SP , DB01221 , or capsaicin in the perfusion medium . 3 . Basal DB00917 levels from spinal cords of animals that underwent 5-72 h of peripheral inflammation were elevated relative to age-matched naive cohorts . The time course of this increase in basal DB00917 levels coincided with peripheral inflammation , as assessed by knee joint circumference . Basal 6-keto-PGF1alpha levels were not elevated after injury . 4 . From this inflammation-evoked increase in basal DB00917 levels , SP and capsaicin significantly increased spinal DB00917 release in a dose-dependent fashion . DB06774 -evoked increases were blocked dose-dependently by inclusion of S(+) ibuprofen in the capsaicin-containing perfusate . 5 . These data suggest a role for spinal DB00917 and P25103 activation in the development of hyperalgesia after injury and demonstrate that this relationship is upregulated in response to peripheral tissue injury and inflammation . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . Cyto- and receptor architecture of area 32 in human and macaque brains . Human area 32 plays crucial roles in emotion and memory consolidation . It has subgenual ( s32 ) , pregenual ( Q9H160 ) , dorsal , and midcingulate components . We seek to determine whether macaque area 32 has subgenual and pregenual subdivisions and the extent to which they are comparable to those in humans by means of NeuN immunohistochemistry and multireceptor analysis of laminar profiles . The macaque has areas s32 and Q9H160 . In s32 , layer IIIa/b neurons are larger than those of layer IIIc . This relationship is reversed in Q9H160 . Layer Va is thicker and Vb thinner in s32 . Area Q9H160 contains higher kainate , benzodiazepine ( BZ ) , and serotonin (5-HT)1A but lower N-methyl-D-aspartate ( DB01221 ) and α2 receptor densities . Most differences were found in layers I , II , and VI . Together , these differences support the dual nature of macaque area 32 . Comparative analysis of human and macaque s32 and Q9H160 supports equivalences in cyto- and receptor architecture . Although there are differences in mean areal receptor densities , there are considerable similarities at the layer level . Laminar receptor distribution patterns in each area are comparable in the two species in layers III-Va for kainate , DB01221 , γ-aminobutyric acid (GABA)B , BZ , and P08908 receptors . Multivariate statistical analysis of laminar receptor densities revealed that human s32 is more similar to macaque s32 and Q9H160 than to human Q9H160 . Thus , macaque 32 is more complex than hitherto known . Our data suggest a homologous neural architecture in anterior cingulate s32 and Q9H160 in human and macaque brains . Anti-stress effect of astragaloside IV in immobilized mice . ETHNOPHARMACOLOGICAL RELEVANCE : Astragaloside IV , a major component extracted from the roots of Astragalus membranaceus ( AM ) , possesses anti-inflammatory , anti-oxidative , anti-fibrotic , anti-infarction and immunoregulatory effects . To clarify anti-stress effect of AM , anxiolytic and anti-inflammatory effects of 80 % ethanol extract of AM and astragaloside IV were investigated in immobilization stress model . MATERIALS AND METHODS : The mice were orally administered with AM ( 50 , 200 , and 500 mg/kg ) , astragaloside IV ( 5 , 10 , and 20 mg/kg ) and buspirone , a positive drug , 1h before immobilization treated for 2h . For anxiolytic activity assay , EPM test was performed in mice . For anti-inflammatory activity assay , serum levels of corticosterone , P05231 and P01375 -α were measured using ELISA kits . RESULTS : AM extract and astragaloside IV increased dose-dependently time spent on open arms and open arm entries in the EPM test . Anxiolytic effects of AM extract ( 500 mg/kg ) and astragaloside IV ( 20 mg/kg ) were comparable to those of buspirone ( 1 mg/kg ) . Their anxiolytic effects were blocked by WAY-100635 ( 0.5 mg/kg , i.p. ) , a P08908 receptor antagonist ( p < 0.01 ) , but not by flumazenil ( 3 mg/kg , i.p. ) and bicuculline ( 0.5 mg/kg , i.p. ) , GABAA receptor antagonists . AM extract and astragaloside IV also reduced serum levels of corticosterone , P05231 and P01375 -α dose-dependently . CONCLUSIONS : AM , particularly astragaloside IV , may ameliorate immobilized stress-induced anxiety and inflammation . Clinical endocannabinoid deficiency ( CECD ) : can this concept explain therapeutic benefits of cannabis in migraine , fibromyalgia , irritable bowel syndrome and other treatment-resistant conditions ? OBJECTIVES : This study examines the concept of clinical endocannabinoid deficiency ( CECD ) , and the prospect that it could underlie the pathophysiology of migraine , fibromyalgia , irritable bowel syndrome , and other functional conditions alleviated by clinical cannabis . METHODS : Available literature was reviewed , and literature searches pursued via the National Library of Medicine database and other resources . RESULTS : Migraine has numerous relationships to endocannabinoid function . Anandamide ( AEA ) potentiates P08908 and inhibits 5- Q13049 receptors supporting therapeutic efficacy in acute and preventive migraine treatment . Cannabinoids also demonstrate dopamine-blocking and anti-inflammatory effects . AEA is tonically active in the periaqueductal gray matter , a migraine generator . THC modulates glutamatergic neurotransmission via DB01221 receptors . Fibromyalgia is now conceived as a central sensitization state with secondary hyperalgesia . Cannabinoids have similarly demonstrated the ability to block spinal , peripheral and gastrointestinal mechanisms that promote pain in headache , fibromyalgia , IBS and related disorders . The past and potential clinical utility of cannabis-based medicines in their treatment is discussed , as are further suggestions for experimental investigation of CECD via P04141 examination and neuro-imaging . CONCLUSION : Migraine , fibromyalgia , IBS and related conditions display common clinical , biochemical and pathophysiological patterns that suggest an underlying clinical endocannabinoid deficiency that may be suitably treated with cannabinoid medicines . Gene-gene interaction analyses between DB01221 receptor subunit and dopamine receptor gene variants and clozapine response . AIMS : To investigate the possible association and gene-gene interaction effects of polymorphisms in DB01221 receptor subunit ( GRIN1 , Q12879 and Q13224 ) and dopamine receptor ( P21728 , P14416 and P35462 ) genes with clozapine response . MATERIALS & METHODS : GRIN1 rs11146020 ( G1001C ) , Q12879 GT-repeat and Q13224 rs10193895 ( G-200T ) polymorphisms were tested for association in a Caucasian ( n = 183 ) and an African-American ( n = 49 ) sample using χ(2) and Q9UNW9 tests . Logistic regression and two-way Q9UNW9 were used to explore gene-gene interaction effects with dopamine receptor gene variants . RESULTS & CONCLUSION : This study does not support the involvement of the DB01221 receptor subunit gene polymorphisms in clozapine response . All tests for an association were negative . Gene-gene interaction analyses however yielded promising leads , including an observed effect between P21728 rs686 and P35462 Ser9Gly polymorphisms on clozapine response ( p = 0.002 ) . Modulation of NMDAR subunit expression by O94759 channels regulates neuronal vulnerability to ischemic cell death . Neuronal vulnerability to ischemia is dependent on the balance between prosurvival and prodeath cellular signaling . In the latter , it is increasingly appreciated that toxic Ca(2+) influx can occur not only via postsynaptic glutamate receptors , but also through other cation conductances . One such conductance , the Transient receptor potential melastatin type-2 ( O94759 ) channel , is a nonspecific cation channel having homology to Q96QT4 , a conductance reported to play a key role in anoxic neuronal death . The role of O94759 conductances in ischemic Ca(2+) influx has been difficult to study because of the lack of specific modulators . Here we used O94759 -null mice ( O94759 (-/-) ) to study how O94759 may modulate neuronal vulnerability to ischemia . O94759 (-/-) mice subjected to transient middle cerebral artery occlusion exhibited smaller infarcts when compared with wild-type animals , suggesting that the absence of O94759 is neuroprotective . Surprisingly , field potentials ( fEPSPs ) recorded during redox modulation in brain slices taken from O94759 (-/-) mice revealed increased excitability , a phenomenon normally associated with ischemic vulnerability , whereas wild-type fEPSPs were unaffected . The upregulation in fEPSP in O94759 (-/-) neurons was blocked selectively by a Q12879 antagonist . This increase in excitability of O94759 (-/-) fEPSPs during redox modulation depended on the upregulation and downregulation of Q12879 - and Q13224 -containing NMDARs , respectively , and on augmented prosurvival signaling via Akt and P29323 pathways culminating in the inhibition of the proapoptotic factor GSK3β . Our results suggest that O94759 plays a role in downregulating prosurvival signals in central neurons and that O94759 channels may comprise a therapeutic target for preventing ischemic damage . Molecular genetics of attention-deficit/hyperactivity disorder : an overview . As heritability is high in attention-deficit/hyperactivity disorder ( ADHD ) , genetic factors must play a significant role in the development and course of this disorder . In recent years a large number of studies on different candidate genes for ADHD have been published , most have focused on genes involved in the dopaminergic neurotransmission system , such as P21917 , P21918 , Q01959 / Q01959 , P09172 , DDC . Genes associated with the noradrenergic ( such as NET1/ P23975 , P08913 , P18825 ) and serotonergic systems ( such as 5-HTT/ P31645 , P28222 , P28223 , Q8IWU9 ) have also received considerable interest . Additional candidate genes related to neurotransmission and neuronal plasticity that have been studied less intensively include P60880 , P43681 , DB01221 , P23560 , P01138 , P20783 , P34130 /5 , P39905 . This review article provides an overview of these candidate gene studies , and summarizes findings from recently published genome-wide association studies ( GWAS ) . GWAS is a relatively new tool that enables the identification of new ADHD genes in a hypothesis-free manner . Although these latter studies could be improved and need to be replicated they are starting to implicate processes like neuronal migration and cell adhesion and cell division as potentially important in the aetiology of ADHD and have suggested several new directions for future ADHD genetics studies . Subdivisions of human parietal area 5 revealed by quantitative receptor autoradiography : a parietal region between motor , somatosensory , and cingulate cortical areas . Brodmann 's area ( BA ) 5 of the human superior parietal cortex occupies a central anatomical position between the primary motor ( BA 4 ) , somatosensory ( area 3b and BA 2 ) , cingulate ( area 23c ) , and superior parietal association cortex ( BA 7 ) . We studied the regional and laminar distributions of the binding sites of 12 different neurotransmitter receptors ( glutamatergic : AMPA , kainate , DB01221 ; GABAergic : GABAA , GABAB ; cholinergic : muscarinic M2 , nicotinic ; adrenergic : alpha1 , alpha2 ; serotoninergic : P08908 , 5-HT2 ; dopaminergic : D1 ) in human postmortem brains by means of quantitative receptor autoradiography , since the structural and functional aspects of human BA 5 are widely unknown , and previous observations have demonstrated characteristic differences in receptor distribution between motor and somatosensory areas . Binding site densities were measured in the cytoarchitectonically defined BA 5 and surrounding regions . Similarities and differences of receptor distribution between cortical areas were studied by cluster analysis of mean binding site densities averaged over all cortical layers , univariate and multivariate statistics , and by density profiles representing laminar receptor distribution patterns . Based on regional heterogeneities of binding site densities and of the cytoarchitecture within BA 5 , we suggest a subdivision into three subareas : medial area 5M , lateral area 5L , and area 5Ci in the region around the cingulate sulcus . BA 5 is therefore a heterogeneous cortical region , comprising three subareas showing receptor expression patterns similar to the adjoining higher order somatosensory , multimodal parietal , or cingulate regions . These findings suggest that human BA 5 constitutes a higher order cortical area , clearly distinct from the primary somatosensory and motor cortex . Synaptic Q12879 and Q13224 containing DB01221 receptors within the superficial dorsal horn activated following primary afferent stimulation . DB01221 receptors are important elements in pain signaling in the spinal cord dorsal horn . They are heterotetramers , typically composed of two Q05586 and two of four GluN2 subunits : Q12879 -2D . Mice lacking some of the GluN2 subunits show deficits in pain transmission yet functional synaptic localization of these receptor subtypes in the dorsal horn has not been fully resolved . In this study , we have investigated the composition of synaptic DB01221 receptors expressed in monosynaptic and polysynaptic pathways from peripheral sensory fibers to lamina I neurons in rats . We focused on DB05875 receptor-expressing ( P25103 + ) projection neurons , critical for expression of hyperalgesia and allodynia . EAB-318 and ( R ) -CPP , Q12879 /B antagonists , blocked both monosynaptic and polysynaptic DB01221 EPSCs initiated by primary afferent activation by ∼90 % . Physiological measurements exploiting the voltage dependence of monosynaptic EPSCs similarly indicated dominant expression of Q12879 /B types of synaptic DB01221 receptors . In addition , at synapses between C fibers and P25103 + neurons , DB01221 receptor activation initiated a secondary , depolarizing current . DB08954 , a Q13224 antagonist , caused modest suppression of monosynaptic DB01221 EPSC amplitudes , but had a widely variable , sometimes powerful , effect on polysynaptic responses following primary afferent stimulation when inhibitory inputs were blocked to mimic neuropathic pain . We conclude that Q13224 subunits are moderately expressed at primary afferent synapses on lamina I P25103 + neurons , but play more important roles for polysynaptic DB01221 EPSCs driven by primary afferents following disinhibition , supporting the view that the analgesic effect of the Q13224 antagonist on neuropathic pain is at least in part , within the spinal cord . Effect of tandospirone , a serotonin-1A receptor partial agonist , on information processing and locomotion in dizocilpine-treated rats . RATIONALE : Augmentation therapy with serotonin-1A receptor ( P08908 ) partial agonists has been suggested to ameliorate psychotic symptoms in patients with schizophrenia . OBJECTIVE AND METHODS : The objective of the present study was to examine the effect of repeated administration of tandospirone ( 0.05 and 5 mg/kg ) on locomotor activity in a novel environment and on sensorimotor gating in rats treated with the N-methyl-D-aspartate receptor antagonist MK-801 , which has been used in animal models of schizophrenia . Furthermore , we sought to determine whether the effect of tandospirone on these behavioural measures is blocked by WAY 100635 ( 0.3 mg/kg ) , a P08908 receptor antagonist , and whether there is an interaction between haloperidol ( 0.1 mg/kg ; a dopamine-D2 receptor antagonist ) and tandospirone . RESULTS : Tandospirone at 5 mg/kg , but not 0.05 mg/kg , decreased locomotor activity in saline or MK-801-treated rats , which were not affected by co-treatment with WAY 100635 . DB00502 decreased locomotion both in saline and MK-801-treated animals , and this effect was not evident in the latter group receiving the higher dose of tandospirone . Tandospirone ( 5 mg/kg ) -induced disruption of sensorimotor gating in saline or MK-801-treated animals was reversed by WAY-100635 , but not by haloperidol . CONCLUSIONS : These findings suggest that behavioural changes induced by tandospirone are not fully blocked by P08908 antagonists and that tandospirone ( 5 mg/kg ) potentiates the effect of MK-801 . Overall , these findings point to an interaction between DB01221 and 5-HT(1A) receptors . Part of the effect of tandospirone on locomotor activity may be mediated by the actions of its active metabolites on other neurotransmitter systems . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Density and distribution of hippocampal neurotransmitter receptors in autism : an autoradiographic study . Neuropathological studies in autistic brains have shown small neuronal size and increased cell packing density in a variety of limbic system structures including the hippocampus , a change consistent with curtailment of normal development . Based on these observations in the hippocampus , a series of quantitative receptor autoradiographic studies were undertaken to determine the density and distribution of eight types of neurotransmitter receptors from four neurotransmitter systems ( GABAergic , serotoninergic [ 5-HT ] , cholinergic , and glutamatergic ) . Data from these single concentration ligand binding studies indicate that the GABAergic receptor system ( 3[H]-flunitrazepam labeled benzodiazepine binding sites and 3[H]-muscimol labeled GABA(A) receptors ) is significantly reduced in high binding regions , marking for the first time an abnormality in the GABA system in autism . In contrast , the density and distribution of the other six receptors studied ( 3[H]-80H-DPAT labeled P08908 receptors , 3[H]-ketanserin labeled 5-HT2 receptors , 3[H]-pirenzepine labled M1 receptors , 3[H]-hemicholinium labeled high affinity choline uptake sites , 3[H]-MK801 labeled DB01221 receptors , and 3[H]-kainate labeled kainate receptors ) in the hippocampus did not demonstrate any statistically significant differences in binding . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . Statins exhibit anticancer effects through modifications of the pAkt signaling pathway . Statins are cholesterol lowering drugs that exhibit antitumor effects in several in vitro and in vivo models , and epidemiological studies indicate that statins prevent cancer . However , the molecular mechanism underlying the effects of statins still needs to be elucidated . We previously demonstrated that single doses of different statins rapidly affect Akt signaling via the purinergic receptor Q99572 . In particular , statins down-regulated nuclear pAkt . Here , we report that long-term treatment of A549 cells with high concentrations of statins ( 15-75 µM ) selects cell sub-populations exhibiting altered P2X receptor expression , signs of increased P60484 activity , enhanced Q6ZVD8 , decreased PI3K p110β and inhibited downstream pAkt signaling . Furthermore , the nuclear accumulation of pAkt in response to insulin was inhibited in selected cells . Statin-selected cells displayed reduced proliferation rate and were more vulnerable to etoposide- and 5-fluorouracil-elicited cytotoxic effects . The stability of a selected phenotype ( 50 µM ) was tested for three weeks in the absence of statins . This resulted in a reversal of some , but not all alterations . Importantly , the truncated nuclear insulin response was retained . We conclude that long-term treatment with high doses of statins selects cells exhibiting stable alterations in insulin-Akt signaling and which are vulnerable to DNA damage . Our studies strengthen the hypothesis that an altered Akt signaling has a role in chemopreventive effects of statins . Development of peptidomimetic ligands of Pro- DB00149 - DB00145 -NH(2) as allosteric modulators of the dopamine D(2) receptor . A variety of stable , small-molecule peptidomimetic ligands have been developed to elucidate the mechanism by which the neuropeptide Pro- DB00149 - DB00145 -NH(2) ( P00747 ) modulates dopaminergic neurotransmission . Photoaffinity labeling ligands based upon P00747 peptidomimetics have been used to establish that P00747 binds to the P14416 at a site that is different from the orthosteric site , thus making P00747 and its peptidomimetics allosteric modulators of the dopamine receptor . Through the design , synthesis and pharmacological evaluation of conformationally constrained peptidomimetics containing lactam , bicyclic , and spiro-bicyclic scaffolds , support was provided for the hypothesis that the bioactive conformation of P00747 is a type II β-turn . In addition , studies with peptidomimetics designed to mimic either a type VI β-turn or polyproline II helix conformation yielded molecules that were able to modulate dopamine receptors because of their ability to place the carboxamide NH(2) pharmacophore in the same topological space as that seen in the type II β-turn . Extensive studies with the spiro-bicyclic P00747 peptidomimetics also established that both positive and negative modes of modulation were possible for the same series of peptidomimetics simply as a result of minor differences in the stereochemistry about the bridgehead carbon within the scaffold . This information was used to transform existing positive modulators into negative modulators , which demonstrated that small structural changes in the spiro-bicyclic dopamine receptor modulators are capable of causing major changes in the modulatory activity of P00747 peptidomimetics . 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 . Neurological impairment in experimental antiphospholipid syndrome is associated with increased ligand binding to hippocampal and cortical serotonergic P08908 receptors . The antiphospholipid syndrome ( APS ) is an autoimmune disease where the presence of high titers of circulating autoantibodies causes thrombosis with consecutive infarcts . In experimental APS ( eAPS ) , a mouse model of APS , behavioral abnormalities develop in the absence of vessel occlusion or infarcts . Using brain hemispheres of control and eAPS mice with documented neurological and cognitive deficits , we checked for lymphocytic infiltration , activation of glia and macrophages , as well as alterations of ligand binding densities of various neurotransmitter receptors to unravel the molecular basis of this abnormal behavior . Lymphocytic infiltrates were immunohistochemically characterized using antibodies against CD3 , P01730 , CD8 and forkhead box P09131 ( Foxp3 ) , respectively . P14136 , Iba1 and P34810 -immunohistochemistry was performed , to check for activation of astrocytes , microglia and macrophages . Ligand binding densities of DB01221 , AMPA , GABAA and P08908 receptors were analyzed by in vitro receptor autoradiography . No significant inflammatory reaction occurred in eAPS mice . There was neither activation of astrocytes or microglia nor accumulation of macrophages . Binding values of excitatory and inhibitory neurotransmitter receptors were largely unchanged . However , ligand binding densities of the modulatory serotonergic P08908 receptors in the hippocampus and in the primary somatosensory cortex of eAPS mice were significantly upregulated which is suggested to induce the behavioral abnormalities observed . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . A negative feedback loop mediated by P40763 limits human Th17 responses . The transcription factor P40763 is critically required for the differentiation of Th17 cells , a T cell subset involved in various chronic inflammatory diseases . In this article , we report that P40763 also drives a negative-feedback loop that limits the formation of Q16552 -producing T cells within a memory population . By activating human memory P01730 (+)CD45RO(+) T cells at a high density ( HiD ) or a low density ( LoD ) in the presence of the pro-Th17 cytokines IL-1β , IL-23 , and TGF-β , we observed that the numbers of Th17 cells were significantly higher under LoD conditions . Assessment of P40763 phosphorylation revealed a more rapid and stronger P40763 activation in HiD cells than in LoD cells . Transient inhibition of active P40763 in HiD cultures significantly enhanced Th17 cell numbers . Expression of the P40763 -regulated ectonucleotidase P49961 , which catalyzes DB00171 hydrolysis , was higher in HiD , than in LoD , cell cultures . Interestingly , inhibition of P49961 ectonucleotidase activity enhanced Th17 responses under HiD conditions . Conversely , blocking the DB00171 receptor Q99572 reduced Th17 responses in LoD cultures . These data suggest that P40763 negatively regulates Th17 cells by limiting the availability of DB00171 . This negative-feedback loop may provide a safety mechanism to limit tissue damage by Th17 cells during chronic inflammation . Furthermore , our results have relevance for the design of novel immunotherapeutics that target the P40763 -signaling pathway , because inhibition of this pathway may enhance , rather than suppress , memory Th17 responses . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . [ Quantitative analysis of P11387 activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin-11 ] . DB00762 ( CPT-11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT-11 was mediated through interaction of the drugs with its target enzyme , P11387 ( topo I ) . In this study , we studied the relation between sensitivity to CPT-11 and topo I activity of glioma cells . Furthermore , we established CPT-11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT-11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT-11 sensitive group ( IC50 values for CPT-11 ; < 50 micrograms/ml ) tended to be higher than those in CPT-11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT-11 resistance cell lines ( T98G/CPT-11 and P13671 ) respectively exhibit a 5.4- and 7.3-fold increase in resistance to CPT-11 . No differences in topo I activity and intracellular accumulation of CPT-11 were observed between parent and CPT-11 resistant lines . On the other hand , topo I from T98G/CPT-11 and P13671 /CPT-11 cells were at least 4- and 2-fold resistant to the inhibitory effect of the CPT-11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT-11 . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . P23560 and basic fibroblast growth factor downregulate DB01221 receptor function in cerebellar granule cells . Evidence has accumulated to suggest that the DB01221 glutamate receptor subtype plays an important role in neuronal degeneration evoked by hypoxia , ischemia , or trauma . Cerebellar granule cells in culture are vulnerable to DB01221 -induced neuronal excitotoxicity . In these cells , brain-derived neurotrophic factor ( P23560 ) and basic fibroblast growth factor ( P09038 ) prevent the excitotoxic effect of DB01221 . However , little is known about the molecular mechanisms underlying the protective properties of these trophic factors . Using cultured rat cerebellar granule cells , we investigated whether P23560 and P09038 prevent DB01221 toxicity by downregulating DB01221 receptor function . Western blot and RNase protection analyses were used to determine the expression of the various DB01221 receptor subunits ( Q9UHB4 , Q12879 , Q13224 , and Q14957 ) after P23560 or P09038 treatment . P09038 and P23560 elicited a time-dependent decrease in the expression of Q12879 and Q14957 subunits . Because DB01221 receptor activation leads to increased intracellular Ca2+ concentration ( [Ca2+]i ) , we studied the effect of the P23560 - and P09038 -induced reduction in Q12879 and Q14957 synthesis on the DB01221 -evoked Ca2+ responses by single-cell fura-2 fluorescence ratio imaging . P23560 and P09038 reduced the DB01221 -mediated [Ca2+]i increase with a time dependency that correlates with their ability to decrease Q12879 and Q14957 subunit expression , suggesting that these trophic factors also induce a functional downregulation of the DB01221 receptor . Because sustained [Ca2+]i is believed to be causally related to neuronal injury , we suggest that P23560 and P09038 may protect cerebellar granule cells against excitotoxicity by altering the DB01221 receptor-Ca2+ signaling via a downregulation of DB01221 receptor subunit expression . Interactome mapping of the phosphatidylinositol 3-kinase-mammalian target of rapamycin pathway identifies deformed epidermal autoregulatory factor-1 as a new glycogen synthase kinase-3 interactor . The phosphatidylinositol 3-kinase-mammalian target of rapamycin ( PI3K- P42345 ) pathway plays pivotal roles in cell survival , growth , and proliferation downstream of growth factors . Its perturbations are associated with cancer progression , type 2 diabetes , and neurological disorders . To better understand the mechanisms of action and regulation of this pathway , we initiated a large scale yeast two-hybrid screen for 33 components of the PI3K- P42345 pathway . Identification of 67 new interactions was followed by validation by co-affinity purification and exhaustive literature curation of existing information . We provide a nearly complete , functionally annotated interactome of 802 interactions for the PI3K- P42345 pathway . Our screen revealed a predominant place for glycogen synthase kinase-3 ( GSK3 ) A and B and the AMP-activated protein kinase . In particular , we identified the deformed epidermal autoregulatory factor-1 ( O75398 ) transcription factor as an interactor and in vitro substrate of P49840 and P49841 . Moreover , GSK3 inhibitors increased O75398 transcriptional activity on the P08908 serotonin receptor promoter . We propose that O75398 may represent a therapeutic target of lithium and other GSK3 inhibitors used in bipolar disease and depression . Attenuated P08908 receptor signaling in brains of suicide victims : involvement of adenylyl cyclase , phosphatidylinositol 3-kinase , Akt and mitogen-activated protein kinase . Positron emission tomography studies in major depression show reduced serotonin (5-HT)1A receptor antagonist-binding potentials in many brain regions including occipital cortex . The functional meaning of this observation in terms of signal transduction is unknown . We used postmortem brain samples from depressed suicide victims to examine the downstream effectors of P08908 receptor activation . The diagnosis was established by means of psychological autopsy using Diagnostic and Statistical Manual of Mental Disorders ( DSM ) III-R criteria . Measurements of [35S]GTPgammaS binding to Galphai/o in the occipital cortex of suicide victims and matched controls revealed a blunted response in suicide subjects and a decrease in the coupling of P08908 receptor to adenylyl cyclase . No significant group differences were detected in the expression levels of Galphai/o , Galphaq/11 or Galphas proteins , or in the activity of DB02527 -dependent protein kinase A . Studies of a parallel transduction pathway downstream from P08908 receptor activation demonstrated a decrease in the activity of phosphatidylinositol 3-kinase and its downstream effector Akt , as well as an increase in P60484 ( phosphatase and tensin homolog deleted on chromosome 10 ) , the phosphatase that hydrolyzes phosphatidylinositol 3,4,5-triphosphate . Finally , the activation of extracellular signal-regulated kinases 1 and 2 was attenuated in suicide victims . These data suggest that the alterations in agonist-stimulated P08908 receptor activation in depressed suicide victims are also manifest downstream from the associated G protein , affecting the activity of second messengers in two P08908 receptor transduction pathways that may have implications for cell survival . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Clozapine inhibits catalepsy induced by olanzapine and loxapine , but prolongs catalepsy induced by P35240 23390 in rats . Loxapine ( 0.3 mg/kg s.c. ) , olanzapine ( 10 mg/kg s.c. ) and P35240 23390 ( R-(+)-chloro-2 , 3 , 4 , 5-tetrahydro-3-methyl-5-phenyl-1-H-3-benzazepine ; 1 mg/kg , s.c. ) , but not clozapine ( 10 mg/kg , s.c. ) , induced catalepsy in rats . Co-administration of clozapine ( 1 , 3 and 10 mg/kg s.c. ) dose-dependently inhibited loxapine-induced catalepsy . Clozapine ( 10 mg/kg s.c. ) also prevented the induction of catalepsy by olanzapine . In addition , clozapine abolished the catalepsy induced by loxapine when it was administered after the response had fully developed . In contrast , the duration of P35240 23390-induced catalepsy was prolonged by clozapine , indicating that its anti-catalepsy effects against olanzapine and loxapine are unlikely to be caused by muscle relaxation , sedation or stimulation . Since P35240 23390-induced catalepsy is reported to be blocked by scopolamine , dizocilpine ( MK-801 ) or 8-hydroxy-dipropylamino-tetralin , it is unlikely that muscarinic blockade , DB01221 ion channel blockade and P08908 receptor agonism , respectively , are involved in clozapine 's action , but the mechanism by which clozapine exerts this anti-cataleptic effect remains unknown . Neurophysiological analysis of circadian rhythm entrainment . We review recent studies in our laboratory that have investigated the neural mechanisms underlying photic entrainment of the mammalian circadian system . The results from studies of extracellular single-unit recordings and of photic induction of Fos-like immunoreactivity ( Fos-lir ) indicate that excitatory amino acid ( EAA ) transmission , and particularly activation of the N-methyl-D-aspartate ( DB01221 ) receptor subtype , is important for conveying photic information to suprachiasmatic nucleus ( SCN ) cells . We have also found that a subregion of the SCN still shows Fos-lir after blockade of EAA receptors , and we have evidence suggesting that these cells are innervated by a distinct subdivision of the retinal projection to the SCN . In addition , we have found that photic responses of cells in the intergeniculate leaflet ( which projects to the SCN ) and of SCN cells are modulated by serotonin ( 5-HT ) via a receptor that resembles the P08908 subtype . Ocular surface injury induces inflammation in the brain : in vivo and ex vivo evidence of a corneal-trigeminal axis . PURPOSE : To test whether a corneal injury can stimulate inflammation in the trigeminal ganglion ( TG ) , a structure located in the brain . METHODS : At 4 and 8 days after alkali burn induced in the right eyes of mice , in vivo magnetic resonance imaging ( Q9BWK5 ) of the brain was done before and after ultrasmall superparamagnetic iron oxide nanoparticle ( USPIO ) contrast to track macrophages . Trigeminal ganglia were stained for Prussian Blue and inflammatory cell markers . Interleukin-1β , P01375 -α , and P15692 transcripts were quantified on days 1 , 4 , and 8 , and 4 days after corneal topical anti-inflammatory treatment with 0.2 % dexamethasone . The expression of Substance P and its receptor P25103 was also measured in the TG on day 4 . RESULTS : Corneal alkali burn induced leukocyte infiltration , including T cells , in the right TG at 4 and 8 days . In vivo Q9BWK5 showed an increased contrast uptake in the right TG , which peaked at day 8 . Prussian Blue(+) USPIO(+) macrophages were observed in the right TG and exhibited an M2 phenotype . The M2-macrophage infiltration was preponderant in the TG after damage . The proinflammatory cytokines Substance P and P25103 were significantly increased in both the TGs . The expression of IL-1β and P15692 was significantly reduced in the right TG with dexamethasone treatment . CONCLUSIONS : We suggest , for the first time , inflammatory involvement of brain structures following ocular surface damage . Our findings support the hypothesis that the neuropeptide Substance P may be involved in the propagation of inflammation from the cornea to the TG through corneal nerves . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . TATA-driven transcriptional initiation and regulation of the rat serotonin P08908 receptor gene . The transcriptional initiation and regulation of the rat serotonin P08908 receptor gene were characterized . By three types of analyses , a single brain-specific site of transcriptional initiation was localized to -967 bp upstream of the translation initiation codon that is utilized both in hippocampus and in the rat raphe RN46A cell line . This major site of transcriptional initiation was located 58 bp downstream from a consensus TATA element , suggesting TATA-driven transcription of the rat P08908 receptor . To identify the promoter activity of the receptor gene , progressive 5' deletions of the -2,719/-117-bp fragment of the P08908 promoter linked to luciferase gene were transfected into P08908 -negative ( pituitary GH4C1 , Q9BTT4 myoblast , and P13671 glioma ) and P08908 -positive ( septal SN-48 and raphe RN46A ) cell lines . Enhancer regions were identified within a fragment between nucleotides -426 and -117 that selectively enhanced transcription in P08908 -positive cells . A nonselective enhancer/promoter that mediated expression in all cell lines was located upstream between -1,519 and -426 bp in a DNA segment containing consensus TATA , CCAAT , SP-1 , and AP-1 elements as well as a poly-GT26 dinucleotide repeat . Strong repression of transcription in all cell lines was conferred by the region upstream of -1,519 bp that contains a 152-bp DNA segment with > 80 % identity to RANTES , tumor necrosis factor-beta , and other immune system genes . Our results indicate that TATA-driven expression of the P08908 receptor is regulated by a novel proximal tissue-specific enhancer region , a nonselective promoter , and an upstream repressor region that is distinct from previously identified neuron-specific repressors . Polymorphisms influencing olanzapine metabolism and adverse effects in healthy subjects . OBJECTIVE : The pharmacokinetics of olanzapine and response to treatment could be affected by polymorphisms in genes coding for drug-metabolizing enzymes , transporters , or receptors . The aim of this study was to identify genetic markers predictive of pharmacokinetics , pharmacodynamics , and adverse effects of olanzapine . METHODS : Sixty-three healthy volunteers receiving a single 5-mg oral dose of olanzapine were genotyped for 39 genetic variants that could be related to the response to olanzapine . All genetic variants were analyzed by PharmaChip , but P14416 Taq1A polymorphism was determined by real-time polymerase chain reaction . Olanzapine was measured using high-performance liquid chromatography combined with tandem mass spectrometry . The relationship of gender and polymorphisms with olanzapine pharmacokinetics , the change in prolactin levels , and the incidence of adverse effects were evaluated by multiple regression analysis . RESULTS : The pharmacokinetics of olanzapine was influenced by polymorphisms in P20815 , P21266 , and Q13224 . P01236 levels were affected by gender and polymorphisms in P14416 and 5- P28223 . Polymorphisms in P11712 , P51580 , P22309 , P08183 , and 5- P28223 were related to some adverse effects of olanzapine . CONCLUSIONS : Several polymorphisms can explain differences in the pharmacokinetics , pharmacodynamics , and safety of olanzapine in healthy subjects . Whether these genetic factors influence the risk of therapeutic failure or tolerability in patients remains to be established . Pattern and pharmacology of propagating epileptiform activity in mouse cerebral cortex . Multiple extracellular recording electrodes were used to study the intra- and interhemispheric spread of stimulus-evoked epileptiform responses in adult mouse neocortical slices . Bath application of 20 microM bicuculline methiodide induced epileptiform activity that propagated at approximately 0.08 m/s over several millimeters in rostro-caudal and medio-lateral direction within the ipsilateral hemisphere and across the corpus callosum to the contralateral hemisphere . A vertical incision from layer II to subcortical regions did not prevent the spread to remote cortical regions , indicating that layer I plays a major role in the lateral propagation of epileptiform activity . The intra- and interhemispheric spread was not influenced by application of an N-methyl-d-aspartate ( DB01221 ) receptor antagonist , but blocked by an antagonist acting at the (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid ( AMPA ) -type glutamate receptor . The potential role of potassium channel activation in controlling the generation or spread of epileptiform activity was tested by applying the potassium channel opener cromakalim and the serotonin type 1A ( P08908 ) receptor agonist (+/-)-8-hydroxydipropylaminotetralin ( 8-OH-DPAT ) to the disinhibited slices . Whereas cromakalim reduced the neuronal excitability and blocked all epileptiform responses , 8-OH-DAPT did not affect the activity pattern . Our results suggest that propagating epileptiform activity in disinhibited neocortical structures is predominantly mediated by activation of AMPA receptors and controllable by activation of a voltage-dependent potassium current . Involvement of dopaminergic receptor signaling in the effects of glutamatergic receptor antagonists on conditioned place aversion induced by naloxone in single-dose morphine-treated rats . A better understanding of the neurochemical mechanisms mediating the aversive consequences of drug withdrawal is important for understanding drug addiction . We previously demonstrated that the inhibitory effect of glutamate receptor antagonists on the conditioned place aversion ( P15085 ) induced by naloxone-precipitated withdrawal after a single morphine exposure could be blocked by dopamine receptor antagonists . Thus , a glutamatergic-dopaminergic interaction may participate in this phenomenon . The current study was undertaken to further characterize this interaction by employing both D(1) ( P35240 23390 ) and D(2) ( raclopride and eticlopride ) dopamine receptor antagonists . The influence of these antagonists on the attenuation of P15085 by MK-801 ( DB01221 receptor antagonist ) , GYKI 52466 ( AMPA receptor antagonist ) , and MCPG ( metabotropic glutamate receptor antagonist ) was determined in rats receiving a single dose of morphine . The dopamine antagonists showed either a significant reversal or a tendency to reverse the effects of MK-801 on P15085 . The effect of GYKI 52466 was also attenuated by the blockade of either D(1) or D(2) receptors . The effect of MCPG , however , was only blocked by D(2) antagonists and not by the D(1) antagonist P35240 23390 . These results add evidence to the hypothesis that a glutamatergic-dopaminergic interaction may be involved in the P15085 induced by naloxone-precipitated withdrawal following a single morphine exposure and suggest that both D(1) and P14416 signaling mechanisms play a role in mediating the aversive aspects of acute dependence . Regulatory interplay of Cockayne syndrome B ATPase and stress-response gene P18847 following genotoxic stress . Cockayne syndrome type B ATPase ( Q03468 ) belongs to the SwItch/ DB02772 nonfermentable family . Its mutations are linked to Cockayne syndrome phenotypes and classically are thought to be caused by defects in transcription-coupled repair , a subtype of DNA repair . Here we show that after UV-C irradiation , immediate early genes such as activating transcription factor 3 ( P18847 ) are overexpressed . Although the P18847 target genes , including dihydrofolate reductase ( P00374 ) , were unable to recover RNA synthesis in Q03468 -deficient cells , transcription was restored rapidly in normal cells . There the synthesis of P00374 mRNA restarts on the arrival of RNA polymerase II and Q03468 and the subsequent release of P18847 from its DB02527 response element/ P39905 target site . In Q03468 -deficient cells P18847 remains bound to the promoter , thereby preventing the arrival of polymerase II and the restart of transcription . Silencing of P18847 , as well as stable introduction of wild-type Q03468 , restores RNA synthesis in UV-irradiated Q03468 cells , suggesting that , in addition to its role in DNA repair , Q03468 activity likely is involved in the reversal of inhibitory properties on a gene-promoter region . We present strong experimental data supporting our view that the transcriptional defects observed in UV-irradiated Q03468 cells are largely the result of a permanent transcriptional repression of a certain set of genes in addition to some defect in DNA repair . Glutamatergic ( N-methyl-D-aspartate receptor ) hypofrontality in schizophrenia : too little juice or a miswired brain ? P14416 blockade has been an obligate mechanism of action present in all medications that effectively treat positive symptoms of schizophrenia ( e.g. , delusions and hallucinations ) and have been approved by regulatory agencies since the 1950s . Blockade of 5-hydroxytryptamine(2A) receptors plays a contributory role in the actions of the second generation of antipsychotic drugs , the so-called atypical antipsychotics . Nevertheless , substantial unmet medical needs remain for the treatment of negative symptoms and cognitive dysfunction . Recognition that dissociative anesthetics block the N-methyl-D-aspartate ( DB01221 ) receptor channel has inspired a search for glutamatergic therapeutic mechanisms because ketamine and phencyclidine are known to induce psychotic-like symptoms in healthy volunteers and exacerbate the symptoms of patients with schizophrenia . Current pathophysiological theories of schizophrenia emphasize that hypofunction of DB01221 receptors at critical sites in local circuits modulate the function of a given brain region or control projections from one region to another ( e.g. , hippocampal-cortical or thalamocortical projections ) . The demonstration that a metabotropic glutamate 2/3 ( mGlu2/3 ) receptor agonist prodrug decreased both positive and negative symptoms of schizophrenia raised hopes that glutamatergic mechanisms may provide therapeutic advantages . In addition to discussing the activation of mGlu2 receptors with mGlu2/3 receptor agonists or mGlu2 receptor positive allosteric modulators ( PAMs ) , we discuss other methods that may potentially modulate circuits with hypofunctional DB01221 receptors such as glycine transporter inhibitors and mGlu5 receptor PAMs . The hope is that by modulating glutamatergic neurotransmission , the dysfunctional circuitry of the schizophrenic brain ( both local circuits and long-loop pathways ) will be improved . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . Serotonin inhibits epileptiform discharge by activation of P08908 receptors in P00915 pyramidal neurons . The anti-epileptiform effect of serotonin was characterized in cellular models of epilepsy using electrophysiological recording techniques . In the bicuculline model , both serotonin ( 20 microM ) and its P08908 agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT , 10 microM ) completely blocked the epileptiform discharge and caused membrane hyperpolarization and reduction in input resistance . These effects were completely antagonized by the P08908 receptor antagonist N-t-butyl-3(4-[2-methoxyphenyl]piperazin-1-yl)-2-phenyl-propanamid e ( WAY 100135 ) ( 10 microM ) . Epileptiform discharge induced by positive current injection was also blocked by serotonin . The presence of WAY 100135 renders serotonin ineffective in the same model . In the bicuculline model , epileptiform discharge blocked by serotonin reappeared and was also intensified when BaCl2 was added to the medium . To rule out the possibility of serotonin-induced hyperpolarization strengthening the inhibitory effect of endogenous Mg2+ on glutamate N-methyl-D-aspartic acid ( DB01221 ) receptor we studied the antiepileptic effect of serotonin in the 0 Mg2+ model . Spontaneous activity and evoked bursts seen with the 0 Mg2+ model were completely blocked by serotonin . WAY 100135 completely antagonized serotonin effects in this model as well . This study provides evidence suggesting that in rat P00915 pyramidal neurons , serotonin can inhibit epileptiform activity in a variety of accepted epilepsy cellular models and that inhibition of epileptiform bursts by serotonin may be mediated by activation of the P08908 receptor subtype . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . DB06016 ( RGH-188 ) , a potent D3/D2 dopamine receptor partial agonist , binds to dopamine D3 receptors in vivo and shows antipsychotic-like and procognitive effects in rodents . We investigated the in vivo effects of orally administered cariprazine ( RGH-188 ; trans-N-{4-[2-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-ethyl]-cyclohexyl}-N',N'-dimethyl-urea ) , a D(3)/ P14416 partial agonist with ∼10-fold preference for the D(3) receptor . Oral bioavailability of cariprazine at a dose of 1mg/kg in rats was 52 % with peak plasma concentrations of 91ng/mL . DB06016 10mg/kg had good blood-brain barrier penetration , with a brain/plasma AUC ratio of 7.6:1 . In rats , cariprazine showed dose-dependent in vivo displacement of [ (3)H ] (+)-PHNO , a dopamine D(3) receptor-preferring radiotracer , in the D(3) receptor-rich region of cerebellar lobules 9 and 10 . Its potent inhibition of apomorphine-induced climbing in mice ( ED(50)=0.27mg/kg ) was sustained for 8h . DB06016 blocked amphetamine-induced hyperactivity ( ED(50)=0.12mg/kg ) and conditioned avoidance response ( CAR ) ( ED(50)=0.84mg/kg ) in rats , and inhibited the locomotor-stimulating effects of the noncompetitive DB01221 antagonists MK-801 ( ED(50)=0.049mg/kg ) and phencyclidine ( ED(50)=0.09mg/kg ) in mice and rats , respectively . It reduced novelty-induced motor activity of mice ( ED(50)=0.11mg/kg ) and rats ( ED(50)=0.18mg/kg ) with a maximal effect of 70 % in both species . DB06016 produced no catalepsy in rats at up to 100-fold dose of its CAR inhibitory ED(50) value . DB06016 0.02-0.08mg/kg significantly improved the learning performance of scopolamine-treated rats in a water-labyrinth learning paradigm . Though risperidone , olanzapine , and aripiprazole showed antipsychotic-like activity in many of these assays , they were less active against phencyclidine and more cataleptogenic than cariprazine , and had no significant effect in the learning task . The distinct in vivo profile of cariprazine may be due to its higher affinity and in vivo binding to D(3) receptors versus currently marketed typical and atypical antipsychotics . The behavioural effects of MK-801 in rats : involvement of dopaminergic , serotonergic and noradrenergic systems . The non-competitive DB01221 receptor antagonist , MK-801 ( dizocilpine ) , induces in rats a characteristic behavioural syndrome with ataxia , stereotypies and hyperlocomotion . At least part of this behavioural syndrome is thought to be related to interactions between glutamatergic and dopaminergic neurotransmission . Based on recent biochemical evidence that serotonin ( 5-HT ) might also be involved in the effects of MK-801 several 5-HT receptor ligands were tested for effects on MK-801-induced behaviours . The P08908 receptor ligands , ipsapirone and NAN-190 , which are known to display antagonist-like properties in functional models of postsynaptic P08908 receptor activity attenuated or blocked the hyperlocomotion and head weaving observed after administration of MK-801 , whereas the 5-HT2 receptor antagonist , ritanserin , was ineffective in this respect . The dopamine receptor antagonist , haloperidol , and the alpha 1-adrenoceptor antagonist , prazosin , also attenuated behaviours induced by MK-801 . In contrast to its effects on stereotypies induced by MK-801 , ipsapirone potentiated rather than attenuated the stereotyped behaviour induced by the dopamine receptor agonist , apomorphine , indicating that antagonism of MK-801-induced stereotypies by ipsapirone may not be related to the dopaminergic system . The data indicate that , in addition to catecholaminergic systems , serotonergic neurotransmission is significantly involved in the mechanisms by which MK-801 alters behaviour in rats . Neuroprotection against N-methyl-D-aspartate-induced excitotoxicity in rat magnocellular nucleus basalis by the P08908 receptor agonist 8-OH-DPAT . The present study reports the neuroprotective efficacy of the P08908 receptor agonists 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and ipsapirone against in vivo excitotoxic neuronal injury . Excitotoxic cell death was induced by injections of N-methyl-D-aspartate ( DB01221 ) in the rat magnocellular nucleus basalis . The neurodegenerative effects were quantified by image analysis of the axonal density of the nucleus basalis projection to the somatosensory cortex visualized with acetylcholinesterase histochemistry . Pretreatment with 8-OH-DPAT -- but not ipsapirone -- 1 h prior to DB01221 infusion showed significant preservation of cortical cholinergic innervation in all doses tested . Furthermore , 8-OH-DPAT exhibited sustained efficacy under homeothermic conditions in which the body temperature was maintained at 36.8 +/- 0.1 degrees C . These data indicate that selective P08908 receptor activation by 8-OH-DPAT protects against DB01221 -induced excitotoxic neuronal damage , probably as a result of P08908 receptor-mediated neuronal hyperpolarization . The novel competitive N-methyl-D-aspartate ( DB01221 ) antagonist CGP 37849 preferentially induces phencyclidine-like behavioral effects in kindled rats : attenuation by manipulation of dopamine , alpha-1 and serotonin1A receptors . The novel competitive N-methyl-D-aspartate ( DB01221 ) receptor antagonist DL-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid ( CGP 37849 ) was found to produce a phencyclidine ( PCP ) -like behavioral syndrome ( ataxia , locomotion , stereotypies ) in amygdala-kindled rats , whereas the amphetamine-like behavioral alterations of the syndrome ( locomotion , stereotypies ) were only infrequently seen in nonkindled rats . In dose-response experiments in kindled and nonkindled rats , behavioral effects were scored using a ranked intensity scale , and the behaviors and behavioural scores determined after CGP 37849 were compared with those determined after i.p. administration of the noncompetitive DB01221 receptor antagonist dizocilpine maleate ( MK-801 ) . In kindled rats , 20 mg/kg of CGP 37849 produced about the same scores for hyperlocomotion and head weaving as 0.1 mg/kg of MK-801 . Kindled rats exhibited higher behavioral scores than nonkindled rats , especially in the case of CGP 37849 . The behavioral effects produced by CGP 37849 in kindled rats were almost indistinguishable from the PCP-like behavioral effects induced by MK-801 , indicating that CGP 37849 indeed produces a PCP-like pattern of behavior in kindled rats . Hyperlocomotion and head weaving induced by CGP 37849 in kindled rats could be attenuated or totally prevented by pretreatment with ipsapirone , a partial agonist/antagonist at postsynaptic 5-hydroxytryptamine ( 5-HT ) receptors of the P08908 subtype . Furthermore , these behavioural effects were attenuated or blocked by the dopamine antagonist haloperidol and the alpha-1 adrenoceptor antagonist , prazosin . The data demonstrate that kindling induces a hypersensitivity to PCP-like behavioral effects of competitive and noncompetitive DB01221 receptor antagonists , which could relate to the recent finding of increased function of DB01221 receptors following kindling. ( ABSTRACT TRUNCATED AT 250 WORDS ) 5-hydroxytryptamine 1A receptor activation protects against N-methyl-D-aspartate-induced apoptotic cell death in striatal and mesencephalic cultures . Apoptosis and glutamate-mediated excitotoxicity may play a role in the pathogenesis of many neurodegenerative disorders , including Parkinson 's disease ( PD ) . In the present study , we investigated whether stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor attenuates N-methyl-D-aspartate- ( DB01221 ) and 1-methyl-4-phenylpyridinium ( P25189 (+) ) -induced apoptotic cell death in cell culture models . A brief exposure ( 20 min ) of M213-2O striatal cells to DB01221 and glutamate produced a delayed increase in caspase-3 activity and DNA fragmentation in a dose- and time-dependent manner . DB01221 -induced caspase-3 activity and DNA fragmentation were almost completely blocked by the P08908 agonists 8-hydroxy-2-(di-n-propylamino)-tetralin ( 8-OH-DPAT ) and ( R ) -5-fluoro-8 hydroxy-2-(dipropylamino)-tetralin ( R-UH-301 ) . Additionally , the protective effects of 8-OH-DPAT and R-UH-301 on DB01221 -induced caspase-3 activation and apoptosis were reversed by pretreatment with the P08908 antagonists N- [ 2-[4-(2-methoxyphenyl)-1-piperazinyl] ethyl ] -N-(2-pyridinyl) cyclohexane carboxamide ( WAY 100635 ) and S-UH-301 , respectively . Similarly , dose- and time-dependent increases in caspase-3 activity and DNA fragmentation were observed in rat primary mesencephalic neurons after a brief exposure to DB01221 and glutamate . P42574 activation and DNA fragmentation in primary mesencephalic neurons were almost completely inhibited by 8-OH-DPAT . This neuroprotective effect of 8-OH-DPAT was reversed by WAY 100635 . Additionally , 8-OH-DPAT blocked tyrosine hydroxylase ( TH ) -positive cell death after DB01221 exposure and also almost completely attenuated the DB01221 -induced Ca(2+) influx in primary mesencephalic cultures . Furthermore , 8-OH-DPAT and R-UH-301 blocked apoptotic cell death in the primary mesencephalic neurons that were exposed to the Parkinsonian toxin P25189 (+) . Together , these results suggest that P08908 receptor stimulation may be a promising pharmacological approach in the development of neuroprotective agents for PD . Habituation in prepulse inhibition is affected by a polymorphism on the DB01221 receptor 2B subunit gene ( Q13224 ) . OBJECTIVES : To identify the reliable connectivity between causal genes or variants with an abnormality expressed in a certain endophenotype has been viewed as a crucial step in unraveling the etiology of schizophrenia because of the considerable heterogeneity in this disorder . METHODS : According to this practical and scientific demand , we aimed to investigate the relationship between seven top-ranked variants in the SZgene database [ 120-bpTR in P21917 , rs1801028 and rs6277 in P14416 , rs1019385 ( T200G ) in Q13224 , rs1800532 in P17752 , rs1801133 ( C677T ) in P42898 , rs2619528 ( P1765 ) in Q96EV8 ] and prepulse inhibition ( PPI ) and habituation after acoustic stimulus ( HAB ) . RESULTS : Both PPI and HAB were decreased significantly in patients with schizophrenia . In addition , we observed a significant effect of Q13224 ( human DB01221 receptor 2B subunit gene , Q13224 ) genotype on HAB ( P < 0.05 , not corrected ) . CONCLUSION : Although these findings need to be replicated in other samples , an underlying mechanism of impaired biological reaction may be influenced by DB01221 hypofunctioning in schizophrenia . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Expression of the receptor-linked protein tyrosine phosphatase P10586 : proteolytic cleavage and shedding of the P62158 -like extracellular region . The human transmembrane molecule P10586 is a protein tyrosine phosphatase ( PTPase ) with a cell adhesion molecule-like extracellular receptor region . The structure of P10586 hinted at its involvement in the regulation of tyrosine phosphorylation through cell-cell or cell-matrix interactions . We show here that P10586 is expressed on the cell surface as a complex of two non-covalently associated subunits derived from a proprotein . The P10586 E-subunit contains the cell adhesion molecule-like receptor region , while the P10586 P-subunit contains a short segment of the extracellular region , the transmembrane peptide and the cytoplasmic PTPase domains . Proprotein processing occurs intracellularly . Analysis of P10586 mutants suggested that cleavage occurs in the P10586 extracellular region at a paired basic amino acid site by a subtilisin-like endoprotease . A single amino acid substitution at this site blocked P10586 proprotein cleavage . The P10586 E-subunit is shed during cell growth , suggesting that P10586 receptor shedding may be a mechanism for regulating PTPase function . The use of immunohistochemistry techniques on human tissues demonstrated the expression of P10586 by various cell lineages , including epithelial cells , smooth muscle cells and cardiac myocytes . The P10586 gene is mapped to chromosome 1 , region Q9H160 -33 , which contains candidate tumor suppressor genes . Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5-linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter ---- MLVI-2 ---- cen ---- P07686 ---- P00374 ---- Pi227- --- cp12.6 ---- ( P05113 , P05112 ) ---- P08700 ---- P04141 ---- P05230 ---- ( P07333 , P09619 ) ---- ( treC,ADRBR ) ---- ( Q5SW96 - Q13585 , P09603 ) ---- qter . The suggested order and orientation for the closely linked P08700 / P04141 gene pair is cen ---- 5' P08700 3' ---- 5' P04141 3' ---- qter , on the basis of analysis of the P04141 rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q- analyses , was telomeric to P04141 and centromeric to P07333 / P09619 , near P05230 . Long-range restriction-enzyme analysis of 5q- DNAs did not detect rearrangements of 5q-linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12.6 , P08700 , and P07333 can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions .

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Transforming growth factor-beta1 induces tumor stroma and reduces tumor infiltrate in cervical cancer . Cervical carcinomas consist of tumor cell nests surrounded by varying amounts of intratumoral stroma containing different quantities and types of immune cells . Besides controlling ( epithelial ) cell growth , the multifunctional cytokine transforming growth factor-beta(1) ( TGF-beta(1) ) is involved in the formation of stroma and extracellular matrix ( Q13201 ) and in immunosuppression . Several malignancies are known to be associated with enhanced production of TGF-beta(1) , repression or mutation of TGF-beta transmembrane receptors , or mutations at the postreceptor intracellular signaling pathway . The aim of our study was to investigate the role of tumor cell-derived TGF-beta(1) on the amount of intratumoral stroma ; the deposition of collagen IV , fibronectin , and laminin ; and the tumor infiltrate in cervical carcinoma . The expression of TGF-beta(1) mRNA in 108 paraffin-embedded cervical carcinomas was detected by mRNA in situ hybridization . Immunohistochemistry was used to investigate the amount of tumor stroma and Q13201 proteins and the extent of the tumor infiltrate . P00747 activator inhibitor-1 ( P05121 ) protein expression in tumor cells was determined to verify the biological activity of TGF-beta(1.) Cytoplasmatic TGF-beta(1) mRNA expression in tumor cells was significantly correlated with the amount of intratumoral stroma and the deposition of collagen IV . TGF-beta(1) mRNA expression in every tumor was accompanied by P05121 expression , indicating biological activity of TGF-beta(1) . An inverse relationship between TGF-beta(1) mRNA expression in tumor cells and the extent of the tumor infiltrate was demonstrated . Our results indicate that cervical cancer cells affect the amount and the composition of the intratumoral stroma and the tumor infiltrate by the production and secretion of TGF-beta(1) . Serotonin activates human monocytes and prevents apoptosis . Monocytes play a critical role in chronic atopic dermatitis ( AD ) and are the primary leukocytes that interact with activated platelets . Although activated platelets release a variety of mediators , the role of platelets in cutaneous allergic inflammation remains unclear . Serotonin ( 5-hydroxytryptamine , 5-HT ) is one of the prototypic mediators produced by activated platelets . We examined the effect of 5-HT on the function and lifespan of human monocytes . Normal human monocytes treated with 5-HT exhibited upregulated expression of costimulatory molecules , enhanced capacity to produce cytokines following lipopolysaccharide treatment , and to stimulate allogeneic P01730 + T cells . 5-HT also attenuated the apoptosis in normal human monocytes in a dose-dependent manner . The plasma levels of 5-HT were increased in patients with AD compared with controls and correlated with the SCORAD index . 5-HT also inhibited monocyte apoptosis in these patients . 5-HT upregulated Bcl-2 and Mcl-1 , and inhibited the activation of caspase-3 . The effects of 5-HT on monocyte apoptosis were mediated by the 5-HT1 and/or P34969 receptors . 5-HT and a 5-HT(1/6/7)-receptor agonist induced phosphorylation of extracellular signal-regulated kinase1/2 and activation of nuclear transcription factor-kappaB . These findings support that 5-HT activates monocytes and inhibits apoptosis , allowing them to remain in the tissue and contribute to chronic inflammation . DB01393 induces plasminogen activator inhibitor-1 gene expression in a O15516 -dependent circadian manner . A functional interaction between peroxisome proliferator-activated receptor alpha ( PPARalpha ) and components of the circadian clock has been suggested , but whether these transcriptional factors interact to regulate the expression of their target genes remains obscure . Here we used a PPARalpha ligand , bezafibrate , to search for PPARalpha-regulated genes that are expressed in a O15516 -dependent circadian manner . Microarray analyses using hepatic RNA isolated from bezafibrate treated-wild type , Clock mutant ( Clk/Clk ) , and PPARalpha-null mice revealed that 136 genes are transcriptionally regulated by PPARalpha in a O15516 -dependent manner . Among them , we focused on the plasminogen activator inhibitor-1 ( P05121 ) gene , because its expression typically shows circadian variation , and it has transcriptional response elements for both Q07869 and O15516 . The bezafibrate-induced expression of P05121 mRNA was attenuated in Clk/Clk mice and in PPARalpha-null mice . The protein levels of PPARalpha were reduced in Clk/Clk hepatocytes . However , the overexpression of PPARalpha could not rescue bezafibrate-induced P05121 expression in Clk/Clk hepatocytes , suggesting that impaired bezafibrate-induced P05121 expression in Clk/Clk mice is not due to reduced PPARalpha expression . Luciferase reporter and chromatin immunoprecipitation analyses using primary hepatocytes demonstrated that DNA binding of both PPARalpha and O15516 is essential for bezafibrate-induced P05121 gene expression . Pull-down assays in vitro showed that both PPARalpha and its heterodimerized partner retinoic acid receptor-alpha can serve as potential interaction targets of O15516 . The present findings revealed that molecular interaction between the circadian clock and the lipid metabolism regulator affects the bezafibrate-induced gene expression . Ras-dependent P29323 activation by the human G(s)-coupled serotonin receptors Q13639 (b) and P34969 (a) . Receptor tyrosine kinases activate mitogen-activated protein ( Q96HU1 ) kinases through Ras , P04049 , and MEK . Receptor tyrosine kinases can be transactivated by G protein-coupled receptors coupling to G(i) and G(q) . The human G protein-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) couple to G(s) and elevate intracellular DB02527 . Certain G(s)-coupled receptors have been shown to activate Q96HU1 kinases through a protein kinase A- and Rap1-dependent pathway . We report the activation of the extracellular signal-regulated kinases ( ERKs ) 1 and 2 ( Q8TCB0 and Q8NFH3 Q96HU1 kinase ) through the human serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in COS-7 and human embryonic kidney HEK293 cells . In transfected HEK293 cells , 5-HT-induced activation of P27361 /2 is sensitive to H89 , which indicates a role for protein kinase A . The observed activation of P27361 /2 does not require transactivation of epidermal growth factor receptors . Furthermore , 5-HT induced activation of both Ras and Rap1 . Whereas the presence of P47736 did not influence the 5-HT-mediated activation of P27361 /2 , the activation of P27361 /2 was abolished in the presence of dominant negative Ras ( RasN17 ) . P27361 /2 activation was reduced in the presence of " dominant negative " Raf1 ( RafS621A ) and slightly reduced by dominant negative B-Raf , indicating the involvement of one or more Raf isoforms . These findings suggest that activation of P27361 /2 through the human G(s)-coupled serotonin receptors 5-HT(4(b)) and 5-HT(7(a)) in HEK293 cells is dependent on Ras , but independent of Rap1 . 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 . DB06288 promotes cognitive flexibility in rats : the role of P34969 receptors . The antagonism of P34969 receptors may contribute to the antidepressant and procognitive actions of the atypical antipsychotic drug , amisulpride . It has been previously demonstrated that the selective P34969 receptor antagonist reversed restraint stress-induced cognitive impairments in a rat model of frontal-dependent attentional set-shifting task ( ASST ) . Therefore , the first aim of the present study was to assess the effectiveness of amisulpride against stress-evoked cognitive inflexibility . The second goal was to elucidate whether the pro-cognitive effect of amisulpride could be due to the compound 's action at P34969 receptors . Rats repeatedly exposed ( 1 h daily for 7 days ) to restraint stress demonstrated impaired performance on the extra-dimensional ( ED ) set-shifting stage of the ASST . DB06288 ( 3 mg/kg ) given to stressed rats 30 min before testing reversed this restraint-induced cognitive inflexibility and improved ED performance of the unstressed control group . The P34969 receptor agonist , AS19 ( 10 mg/kg ) , abolished the pro-cognitive efficacy of amisulpride ( 3 mg/kg ) . The present study suggests that the antagonism of P34969 receptors may contribute to the mechanisms underlining the pro-cognitive action of amisulpride . These results may have therapeutic implications in frontal-like deficits associated with stress-related disorders . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . Effects of lurasidone on executive function in common marmosets . Cognitive impairment is one of the major symptoms of schizophrenia , and is considered largely due to dysfunctions in the prefrontal cortex ( P27918 ) . DB08815 , a novel atypical antipsychotic agent with high binding affinity for dopamine D2 , serotonin P34969 , 5- Q13049 and P08908 receptors has been reported to have superior efficacy in rodents ' models of cognitive impairment . However , the beneficial effect of lurasidone on cognitive impairment has not been evaluated in non-human primates . In this study , we investigated the effect of lurasidone on executive function , which is one of the cognitive domains , in common marmosets and compared the results to those of other antipsychotics . The effects of lurasidone , haloperidol , olanzapine , risperidone , quetiapine and clozapine on executive function were evaluated in naïve marmosets using the object retrieval with detours ( ORD ) task . Before drug treatment , marmosets ' success rates in the easy trial of the test were almost 90 % . However , maximum success in the difficult trial of the task reached only 50 % after 8 days of training . DB00502 , olanzapine and risperidone decreased correct performance even in the easy trial of the task . All drugs , except lurasidone , impaired success rate in the difficult trial . On the other hand , lurasidone dose-dependently increased marmosets ' success rates in the difficult trial with significant effect at 10mg/kg . In conclusion , we have shown in this study that lurasidone , unlike conventional antipsychotics , improves cognition associated with executive function in common marmosets . These findings suggest that lurasidone would be more useful for treatment of schizophrenia cognitive impairment than other antipsychotics . Comparison of the effects of aging on P34969 and P08908 receptors in discrete regions of the circadian timing system in hamsters . The circadian timekeeping system exhibits many functional changes with aging , including a loss of sensitivity to time cues such as systemic injections of the serotonergic agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) . In order to elucidate the neurochemical mechanisms responsible for this age-related loss of sensitivity of the circadian pacemaker to serotonin agonists , the present study used quantitative autoradiography to determine whether aging decreases serotonin receptor populations in male Syrian hamsters . Four neuroanatomical regions that regulate circadian timekeeping were studied ( the suprachiasmatic nuclei [ SCN ] , the lateral geniculate nuclei [ P81274 ] , and the median raphe nucleus [ MRN ] and dorsal raphe nucleus [ DRN ] ) . The specific binding of [3H]8-OH-DPAT to serotonin7 ( P34969 ) and serotonin1A ( P08908 ) receptors was investigated by competitive inhibition with ritanserin and pindolol , respectively . The results showed that the SCN , IGL , MRN , and DRN of the male Syrian hamster exhibited specific binding of [3H]8-OH-DPAT to both the P34969 and P08908 receptors , and that the latter receptor subtype is more abundant in all of these regions . At 17-19 months of age , a 50 % decrease in P34969 receptors was found in the DRN but not in any other regions . No significant age-related changes in P08908 receptors were observed in any regions examined . The finding that a marked decrease in P34969 receptors occurs in the DRN at the age previously characterized by loss of sensitivity to 8-OH-DPAT suggests that this region and this receptor subtype play important roles in 8-OH-DPAT induction of circadian phase shifts in vivo and that they constitute an important locus of aging in the circadian timing system . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Role of monoamine oxidases in the exaggerated 5-hydroxytryptamine-induced tension development of human isolated preeclamptic umbilical artery . We investigated the role(s) of monoamine oxidases ( MAOs ) on the altered 5-hydroxytryptamine ( 5-HT , serotonin ) -induced tension development of the isolated umbilical artery of preeclamptic pregnancy of Chinese women . An enhanced 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy was observed when compared with that of normal pregnancy . The enhanced component of 5-HT-induced tension development was eradicated by clorgyline ( a P21397 inhibitor ) . Blockade of P29474 ( endothelial isoform nitric oxide synthase ) ( N(omega)-nitro-L-arginine methyl ester ) , 5-HT transporter ( citalopram ) , 5-HT receptor subtypes ( 5HT2B , SB 204741 ; P28335 , RS 102221 ; P34969 , SB 269970 ) , and endothelium denudation of the umbilical artery of normal pregnancy mimicked the enhanced 5-HT-induced tension development as observed in the preeclamptic tissues . In contrast , no apparent changes in 5-HT-induced tension development of the umbilical artery of preeclamptic pregnancy were observed with the same pharmacological manipulations . A decreased protein expression levels of P21397 and P29474 ( no P35228 and P27338 expression was detected ) and no change in caveolin-1 and 5-HT transporter expression were demonstrated in the umbilical artery ( endothelium intact ) lysate of preeclamptic pregnancy , compared to that of the umbilical artery of normal pregnancy . Thus , in the umbilical artery of preeclamptic pregnancy , a decrease of P21397 and P29474 protein expression levels are probably associated with , or responsible for , the exaggerated 5-HT-induced tension development . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Two-dimensional liquid crystalline growth within a phase-field-crystal model . By using a two-dimensional phase-field-crystal ( P27918 ) model , the liquid crystalline growth of the plastic triangular phase is simulated with emphasis on crystal shape and topological defect formation . The equilibrium shape of a plastic triangular crystal ( PTC ) grown from an isotropic phase is compared with that grown from a columnar or smectic-A ( Q13216 ) phase . While the shape of a PTC nucleus in the isotropic phase is almost identical to that of the classical P27918 model , the shape of a PTC nucleus in Q13216 is affected by the orientation of stripes in the Q13216 phase , and irregular hexagonal , elliptical , octagonal , and rectangular shapes are obtained . Concerning the dynamics of the growth process , we analyze the topological structure of the nematic order , which starts from nucleation of +1/2 and -1/2 disclination pairs at the PTC growth front and evolves into hexagonal cells consisting of +1 vortices surrounded by six satellite -1/2 disclinations . It is found that the orientational and the positional order do not evolve simultaneously ; the orientational order evolves behind the positional order , leading to a large transition zone , which can span over several lattice spacings . Promotion of colorectal cancer invasion and metastasis through activation of NOTCH- O75553 - P00519 -RHOGEF protein O75962 . We have recently identified a metastasis suppressor gene for colorectal cancer : Q08117 /Aes , which encodes an endogenous inhibitor of NOTCH signaling . When Aes is knocked out in the adenomatous epithelium of intestinal polyposis mice , their tumors become malignant , showing marked submucosal invasion and intravasation . Here , we show that one of the genes induced by NOTCH signaling in colorectal cancer is O75553 /Dab1 . Genetic depletion of O75553 suppresses cancer invasion and metastasis in the NOTCH signaling-activated mice . O75553 is phosphorylated by P00519 tyrosine kinase , which activates P00519 reciprocally . Consistently , inhibition of P00519 suppresses cancer invasion in mice . Furthermore , we show that one of the targets of P00519 is the P31749 /RHOGEF protein O75962 , and that phosphorylation at its DB00135 residue 2681 ( pY2681 ) causes P08100 activation in colorectal cancer cells . Its unphosphorylatable mutation O75962 Y2681F reduces RHOGEF activity and inhibits invasion of colorectal cancer cells . Importantly , O75962 pY2681 correlates with significantly poorer prognosis of patients with colorectal cancer after surgery . SIGNIFICANCE : These results indicate that O75962 pY2681 is one of the downstream effectors of NOTCH signaling activation in colorectal cancer , and can be a prognostic marker , helping to determine the therapeutic modality of patients with colorectal cancer . The influence of costimulation and regulatory P01730 + T cells on intestinal IgA immune responses . It is thought that IgA B-cell differentiation is highly dependent on activated P01730 + T cells . In particular , cell-cell interactions in the Peyer 's patches involving P25942 and/or P33681 / P42081 have been implicated in germinal-center formation and IgA B-cell development . Also soluble factors , such as P05112 , P05113 , P05231 , and TGF beta may be critical for IgA B-cell differentiation in vivo . Here we report on some paradoxical findings with regard to IgA B-cell differentiation and specific mucosal immune responses that we have recently made using gene knockout mice . More specifically , we have investigated to what extent absence of P01730 + T cells , relevant cytokines , or T-cell-B-cell interactions would influence IgA B-cell differentiation in vivo . Using P01730 - or P05112 -gene knockout mice or mice made transgenic for DB01281 , we found that , although specific responses were impaired , total IgA production and IgA B-cell differentiation appeared to proceed normally . However , a poor correlation was found between , on the one hand , GC formation and IgA differentiation and , on the other hand , the ability to respond to T-cell-dependent soluble protein antigens in these mice . Thus , despite the various deficiencies in P01730 + T-cell functions seemingly intact IgA B-cell development was observed . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Serotonin skews human macrophage polarization through P41595 and P34969 . Besides its role as a neurotransmitter , serotonin ( 5-hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT(1-7) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 production , upregulated the expression of M2 polarization-associated genes ( P05120 , P07996 , Q9NY15 , Q86Y22 ) , and reduced the expression of M1-associated genes ( P08476 , P41597 , P39900 , P05121 , P29016 , O94788 ) . Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2( P09603 ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7) , whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies . DB06288 is a potent P34969 antagonist : relevance for antidepressant actions in vivo . RATIONALE : DB06288 is approved for clinical use in treating schizophrenia in a number of European countries and also for treating dysthymia , a mild form of depression , in Italy . DB06288 has also been demonstrated to be an antidepressant for patients with major depression in many clinical trials . In part because of the selective D(2)/D(3) receptor antagonist properties of amisulpride , it has long been widely assumed that dopaminergic modulation is the proximal event responsible for mediating its antidepressant and antipsychotic properties . OBJECTIVES : The purpose of these studies was to determine if amisulpride 's antidepressant actions are mediated by off-target interactions with other receptors . MATERIALS AND METHODS : We performed experiments that : ( 1 ) examined the pharmacological profile of amisulpride at a large number of central nervous system ( CNS ) molecular targets and , ( 2 ) after finding high potency antagonist affinity for human 5-HT(7a) serotonin receptors , characterized the actions of amisulpride as an antidepressant in wild-type and 5-HT(7) receptor knockout mice . RESULTS : We discovered that amisulpride was a potent competitive antagonist at 5-HT(7a) receptors and that interactions with no other molecular target investigated in this paper could explain its antidepressant actions in vivo . Significantly , and in contrast to their wild-type littermates , 5-HT(7) receptor knockout mice did not respond to amisulpride in two widely used rodent models of depression , the tail suspension test and the forced swim test . CONCLUSIONS : These results indicate that 5-HT(7a) receptor antagonism , and not D(2)/D(3) receptor antagonism , likely underlies the antidepressant actions of amisulpride . Serotonin P34969 receptor is critically involved in acute and chronic inflammation of the gastrointestinal tract . BACKGROUND : Intestinal inflammation is often associated with an increased level of serotonin ( 5-HT ) , an important gastrointestinal signaling molecule involved in gut homeostasis through stimulation of specific receptors . In this study , we investigated the role of P34969 receptor ( 5-HT7R ) in the induction and development of intestinal inflammation using a mouse model of acute and chronic colitis and human patients with Crohn 's disease ( CD ) . METHODS : Acute colitis was induced through administration of dextran sodium sulfate to wild-type , 5-HT7R-deficient mice and hematopoietic bone marrow chimera . Chronic colitis was induced in interleukin 10-deficient mice . The role of 5-HT7R in gut inflammation was assessed using agonist/antagonist treatment . We investigated expression and distribution of 5-HT7R , extent of gut inflammation with magnetic resonance imaging and histological analysis , survival rate , and disease activity index . Finally , biopsies from the large intestine of patients with CD were analyzed . RESULTS : Under basal conditions , 5-HT7R is expressed both in enteric neurons and CD11c cells of the large intestine . Expression of 5-HT7R significantly increased after induction of colitis in mice and in inflamed intestinal regions of patients with CD in CD11c/ P42081 double-positive cells . Pharmacological blockade or genetic ablation of 5-HT7R resulted in increased severity of both acute and chronic dextran sodium sulfate-induced colitis , whereas receptor stimulation showed an anti-inflammatory effect . Analysis of bone marrow chimera indicated importance of 5-HT7R expressed by hematopoietic cells in intestinal inflammation . CONCLUSIONS : The 5-HT7R expressed on CD11c/ P42081 -positive myeloid cells modulates the severity of intestinal inflammation in an acute and chronic colitis and thus represents a potential therapeutic target for the treatment of inflammatory disorders such as CD .

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DB11320 influences body temperature by acting at H1 and H3 receptors on distinct populations of preoptic neurons . The preoptic area/anterior hypothalamus , a region that contains neurons that control thermoregulation , is the main locus at which histamine affects body temperature . Here we report that histamine reduced the spontaneous firing rate of GABAergic preoptic neurons by activating H3 subtype histamine receptors . This effect involved a decrease in the level of phosphorylation of the extracellular signal-regulated kinase and was not dependent on synaptic activity . Furthermore , a population of non-GABAergic neurons was depolarized , and their firing rate was enhanced by histamine acting at H1 subtype receptors . In our experiments , activation of the P35367 receptors was linked to the P98160 pathway and Ca(2+) release from intracellular stores . This depolarization persisted in TTX or when fast synaptic potentials were blocked , indicating that it represents a postsynaptic effect . Single-cell reverse transcription-PCR analysis revealed expression of H3 receptors in a population of GABAergic neurons , while H1 receptors were expressed in non-GABAergic cells . DB11320 applied in the median preoptic nucleus induced a robust , long-lasting hyperthermia effect that was mimicked by either H1 or H3 histamine receptor subtype-specific agonists . Our data indicate that histamine modulates the core body temperature by acting at two distinct populations of preoptic neurons that express H1 and H3 receptor subtypes , respectively . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . DB00644 specific binding sites in uterine leiomyomata . DB00644 ( DB00644 ) analogs can cause regression of uterine leiomyomata . This effect is thought to be mediated by the inhibition of gonadotropin release and steroid synthesis . In the present study we examined the possibility that these analogs may also act directly on uterine leiomyomata . Specific binding sites for DB00644 are present in myoma membranes , as 125I- DB06719 binding was displaced with equal efficiency by the superagonists , DB06719 and D-Trp6- DB00644 , and by the antagonist Organon 30276 , but not by unrelated peptides such as thyrotropin releasing hormone and oxytocin . A nonlinear Scatchard curve obtained for DB06719 specific binding suggests the presence of at least two binding sites , one of which exhibits a relatively high affinity for DB00644 analogs ( Kd of approximately 10(-8) M ) . Western blotting with a specific P30968 antibody revealed the presence of a 60 kDa protein in myoma membranes . This protein has a similar molecular weight to the purified pituitary P30968 . These results indicate , for the first time , the presence of specific binding sites for DB00644 in uterine leiomyomata , suggesting a direct effect of DB00644 analogs on this tissue . Development of a yeast protein fragment complementation assay ( DB11245 ) system using dihydrofolate reductase ( P00374 ) with specific additives . A yeast protein fragment complementation assay ( DB11245 ) system based on dihydrofolate reductase ( P00374 ) is difficult to be operated because it is not as sensitive to trimethoprim ( P54849 ) as the system using a prokaryotic microorganism . Here , the DB11245 system using P00374 , specific inhibitors , and a substrate in the yeast Saccharomyces cerevisiae was newly developed . As a model , the human oncoprotein Ras and the Ras-binding domain ( RBD ) of P04049 were individually and genetically fused to P00374 fragment , and each genetic construct was coexpressed under the control of the P22466 promoter . An interaction between Ras and RBD could be evaluated on the basis of cell proliferation . To establish the experimental conditions for the yeast DB11245 system based on the P00374 reconstitution , we examined yeast host strains and the concentration of inhibitory additives to prevent endogenous P00374 activity , namely , P54849 and sulfanilamide , and the substrate of P00374 , namely , folic acid . The transformant harboring wild-type Ras or its variants showed positive interaction signals , and the order of interactions for combination corresponded to the results of other in vitro assays . Moreover , combinatorial mutated Ras-binding domains were constructed , and the interaction of RBDs with Ras using this yeast DB11245 system was examined . As a result , various types of mutated clone for RBD were obtained . These demonstrations suggest that the yeast DB11245 system based on P00374 can be one of good , convenient , and inexpensive tools for investigating eukaryotic protein-protein interactions in vivo . 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 . Expression of gonadotropin-releasing hormone receptor and effect of gonadotropin-releasing hormone analogue on proliferation of cultured gastric smooth muscle cells of rats . AIM : To investigate the expression of gonadotropin-releasing hormone ( DB00644 ) receptor and the effects of DB00644 analog ( alarelin ) on proliferation of cultured gastric smooth muscle cells ( GSMC ) of rats . METHODS : Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of P30968 in GSMC , respectively . Techniques of cell culture , OD value of MTT test , measure of (3)H-TdR incorporation , average fluorescent values of proliferating cell nuclear antigen ( P12004 ) and flow cytometric DNA analysis were used in the experiment . RESULTS : The cultured GSMC of rats showed immunoreactivity for P30968 ; positive staining was located in cytoplasm . P30968 mRNA hybridized signals were also detected in cytoplasm . When alarelin ( 10(-9) , 10(-7) , 10(-5) mol/L ) was administered into the medium and incubated for 24 h , OD value of MTT , (3)H-TdR incorporation and average fluorescent values of P12004 all decreased significantly as compared with the control group ( P < 0.05 ) . The maximum inhibitory effect on cell proliferation was achieved a concentration of 10(-5) mol/L and it acted in a dose-dependent manner . Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G(1) phase and decrease ratio of S phase of GSMC of rats ( P < 0.05 ) . The maximum inhibitory effect on ratio of S phase was at the concentration of 10(-5) mol/L and also acted in a dose-dependent manner . CONCLUSION : Our data suggest that P30968 can be expressed by GSMC of rats . DB00644 analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through DB00644 receptors . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Epigenetic mechanisms in the dopamine D2 receptor-dependent inhibition of the prolactin gene . Transcription of the prolactin gene is dynamically controlled by positive and negative hormone signals that target the regulatory promoter region . Based on the inducibility of prolactin gene expression by inhibitors of histone deacetylases ( HDACs ) , we examined the role of histone acetylation at the genomic prolactin promoter as a late step in transcriptional regulation . Chromatin immunoprecipitation analysis of GH4 cells revealed elevated levels of acetylated histones in the promoter and enhancer regions of the gene , compared with downstream intron sequences . 17beta-Estradiol stimulated histone H4 acetylation in the promoter region by 2- to 3-fold within 30 min . Dopamine inhibited histone H4 acetylation by 2-fold in 30 min , an effect mimicked by the MAPK kinase ( Q02750 ) inhibitor U0126 . In contrast , the synthetic glucocorticoid dexamethasone , which inhibits prolactin transcription , failed to alter histone acetylation over the same time frame . Association of transcription activator Pit-1 with the prolactin promoter was unchanged by hormone treatment . However , in response to dopamine , histone deacetylase Q92769 and corepressor mSin3A were rapidly recruited to the prolactin promoter , and association was sustained above basal levels over a 1-h period . Consistent with this corepressor function , depletion of endogenous mSin3A by small interfering RNA was sufficient to enhance prolactin gene expression by 70 % , comparable to the induction by the HDAC inhibitor , trichostatin A . These studies demonstrate that dopamine D2 receptor activation and inhibition of MAPK ( P27361 /2 ) signaling lead to rapid deacetylation of histones at the genomic prolactin promoter . Recruitment of specific HDAC/ corepressor complexes may be an important mechanism for repression of target gene transcription by Gi/o-coupled receptors . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . P30968 concentration differentially regulates intracellular signaling pathways in GGH3 cells . Pituitary cell lines ( GGH3 ) expressing the P30968 ( GnRHR ) were used to investigate the effect of GnRHR concentration on the ability of a DB00644 agonist to activate second messenger systems . Four different strategies were utilized to generate cells expressing functionally different concentrations of receptors : ( 1 ) transient transfection with different concentrations of wild type GnRHR into GH3 cells , ( 2 ) utilization of two cell lines derived from a common stably transfected line expressing high ( 4,209 +/- 535 receptors/cell ) or low ( 1,031 +/- 36 receptors/cell ) concentrations of GnRHR , ( 3 ) co-incubation of GGH3-1 ' cells with a DB00644 agonist ( DB06719 ) and a DB00644 antagonist to compete for binding sites , and ( 4 ) photo-affinity binding to GnRHR with a DB00644 antagonist to change effective receptor concentration . A range of receptor concentrations ( 1,000-8,000 receptors/cell ) were generated by these techniques . Inositol phosphate ( IP ) and DB02527 accumulation were quantified to assess the effect of receptor concentration on receptor-effector coupling . Under all four paradigms , the efficacy and potency of DB06719 stimulated IP production was dependent on receptor concentration . In contrast , DB06719 stimulated DB02527 release was relatively unchanged at varying concentrations of GnRHR . This suggests that the cellular concentration of GnRHR affects the induction of cell signaling pathways . These results demonstrate that a single ligand-receptor-complex can differentially activate second messenger systems and present a mechanism by which multiple physiological endpoints can be differentially regulated by a single hormone/receptor interaction . Effects of Asn318 and Asp87Asn318 mutations on signal transduction by the gonadotropin-releasing hormone receptor and receptor regulation . P30968 ( P30968 ) contains Asn87 and Asp318 instead of the more frequently observed Asp87 and Asn318 found in other G protein-coupled receptors . In the present study , site-directed mutagenesis was used to introduce Asn318 and Asp87Asn318 into P30968 . The effect on coupling and regulation of P30968 was studied by stable expression of wild and mutant mouse P30968 in the lactotropic GH3 cells ; these normally release PRL in response to TRH stimulation . The responses to DB06719 ( a metabolically stable DB00644 analog ) in three different cell lines , M1 , N8 , and P03886 ( expressing wild-type , Asn318 mutant , and Asp87Asn318 mutant mouse P30968 , respectively ) were compared with that observed in the previously characterized GGH3-1 ' cells , which stably express rat P30968 . The Asn318 and Asp87Asn318 mutations had no measurable effect on ligand binding , but abolished the initial down-regulation of receptor that was observed in M1 and GGH3-1 ' cells , suggesting that the normal location of Asn87 and Asp318 in P30968 is involved in the regulation of P30968 . In N8 and P03886 cells , DB06719 -stimulated inositol phosphate ( IP ) production was attenuated , but the release of both DB02527 and PRL was stimulated in a dose- and time-dependent manner . These mutations apparently impaired the coupling between P30968 and G proteins involved in IP production , but not those involved in DB02527 release . In M1 cells , DB06719 stimulation produced a significant increase in IP production , but neither DB02527 nor PRL release was significantly stimulated . These findings are consistent with the previous suggestion that DB00644 -stimulated PRL release is mediated by a DB02527 second messenger system in transfected GGH3 cells . Dissociable fronto-striatal effects of dopamine D2 receptor stimulation on cognitive versus motor flexibility . Genetic and pharmacological studies suggest an important role of the dopamine D2 receptor ( P14416 ) in flexible behavioral adaptation , mostly shown in reward-based learning paradigms . Recent evidence from imaging genetics indicates that also intentional cognitive flexibility , associated with lateral frontal cortex , is affected by variations in P14416 signaling . In the present functional magnetic resonance imaging ( Q9BWK5 ) study , we tested the effects of a direct pharmacological manipulation of P14416 stimulation on intentional flexibility in a task-switching context , requiring switches between cognitive task rules and between response hands . In a double blind , counterbalanced design , participants received either a low dose of the P14416 agonist bromocriptine or a placebo in two separate sessions . DB01200 modulated the blood-oxygen-level-dependent ( BOLD ) signal during rule switching : rule-switching-related activity in the left posterior lateral frontal cortex and in the striatum was increased compared to placebo , at comparable performance levels . Fronto-striatal connectivity under bromocriptine was slightly increased for rule switches compared to rule repetitions . Hand-switching-related activity , in contrast , was reduced under bromocriptine in sensorimotor regions . Our results provide converging evidence for an involvement of P14416 signaling in fronto-striatal mechanisms underlying intentional flexibility , and indicate that the neural mechanisms underlying different types of flexibility ( cognitive vs motor ) are affected differently by increased dopaminergic stimulation . Q92993 and Q8WYB5 are in positive regulation on cell proliferation of prostate cancer through PI3K-AKT signaling . Histone modifications play important roles in the tumorigenesis and progression of prostate cancer ( PCa ) and genes involved in histone modifications are seemed as ideal targets for treatment of PCa patients . However , clinical trials have shown that those existing drugs exert the minimal antitumor activity and excess adverse effects on PCa patients . Therefore , it is of great interest to figure out novel specific biomarkers to guide the development of new drugs . In present study , an RNAi screening with 44 genes involved in histone modifications was applied to a PCa cell line , Du145 . The results showed that nine genes were in positive regulation of Du145 cell growth . Then four selected genes ( Q92831 , Q92993 , Q8WYB5 and Q13547 ) were found to exert this effect by a gene-specific manner when silenced . And then Q92993 or Q8WYB5 silenced cells were subjected to DNA microarray analysis . The common differentially expressed genes were analyzed by Ingenuity Pathway Analysis ( IPA ) and found that O95238 signaling , EIF2 signaling and PI3K signaling was suppressed following by Q92993 or Q8WYB5 silencing . Subsequent immunoblotting assay showed that AKT signaling was inhibited , which suggested that Q92993 or Q8WYB5 regulates cancer cell growth through PI3K-AKT signaling . Together with our published data [ 31 ] that O14965 inhibitoin increased drug sensitivity of DU145 , our work demonstrated the underlying mechanism that how the acetylation enzyme regulates cancer cells survial and might provide potential therapeutic targets for prostate cancer patients in future epigenetic drug development . Microarray Analysis of the Major Depressive Disorder mRNA Profile Data . OBJECTIVE : Major depressive disorder ( MDD ) is a common mood disorder associated with several psychophysiological changes like disturbances of sleep , appetite , or sexual desire , and it affects the patients ' life seriously . We aimed to explore a genetic method to investigate the mechanism of MDD . METHODS : The mRNA expression profile ( GSE53987 ) of MDD was downloaded from Gene Expression Omnibus database , including 105 samples of three brain regions in post-mortem tissue suffered from MDD and unaffected controls . Differentially expressed genes ( DEGs ) in MDD were identified using the Limma package in R . Gene Ontology functions and Kyoto Enrichment of Genes and Genomes pathways of the selected DEGs were enriched using Database for Annotation , Visualization and Integrated Discovery . Protein-protein interactive network of DEGs was constructed using the Cytoscape software . RESULTS : Totally , 241 DEGs in MDD-hip group , 218 DEGs in MDD-pfc group , and 327 DEGs in MDD-str group were identified . Also , different kinds of biological processes of DEGs in each group were enriched . Besides , glycan biosynthesis of DEGs in MDD-str group , O95786 -like receptor signaling and pyrimidine metabolism of DEGs in the MDD-hip group were enriched , respectively . Moreover , several DEGs like Q05397 , Q13569 and P41208 in MDD-str group , P40126 , AR and P30968 in MDD-pfc group , and P31749 and P51617 in MDD-hip group were selected from PPI network . CONCLUSION : Our data suggests that the brain striatum tissue may be greatly affected by MDD , and DEGs like Q05397 , Q10471 and Q10471 in striatum , AR in prefrontal cortex and P51617 and P29459 in hippocampus may provide novel therapeutic basis for MDD treatment . 37-kDa laminin receptor precursor mediates DB00644 -II-induced P08253 expression and invasiveness in ovarian cancer cells . DB00644 -II enhances ovarian cancer cell invasion in an autocrine manner . We have now found that DB00644 -II increases 37-kDa laminin receptor precursor ( Q14764 ) production in P30968 ( GnRHR ) -positive OVCAR-3 and CaOV-3 ovarian cancer cells , while small interfering RNA ( siRNA ) -mediated depletion of DB00644 -II or GnRHR mRNA abrogates this . The invasiveness of ovarian cancer cells is also reduced > 85 % by siRNA-mediated knockdown of Q14764 levels and > 50 % by pretreatment of Matrigel with a synthetic peptide that blocks interactions between laminin and the 67-kDa nonintegrin laminin receptor which comprises two Q14764 subunits . Conversely , overexpressing Q14764 in CaOV-3 cells increases their invasiveness 5-fold , while overexpressing Q14764 with a nonfunctional laminin-binding site does not . Depletion of Q14764 by siRNA treatment reduces CaOV-3 cell attachment to laminin-coated plates by ∼80 % but only reduces their binding to Matrigel by ∼20 % . Thus , while Q14764 influences CaOV-3 cell adhesion to laminin , Q14764 must act in other ways to enhance invasion . Matrix metalloproteinases ( MMPs ) are key mediators of invasion , and Q14764 siRNA treatment of OVCAR-3 and CaOV-3 cells inhibits P08253 but not P14780 mRNA levels . Overexpressing Q14764 in these cells increases P08253 production specifically , while a laminin-binding deficient Q14764 does not . Importantly , Q14764 siRNA treatment abolishes DB00644 -II-induced P08253 production , and invasion in OVCAR-3 and CaOV-3 cells , which was also seen after P08253 siRNA treatment . These results suggest that DB00644 -II-induced Q14764 expression increases the amount of the 67-kDa nonintegrin laminin receptor , which appears to interact with laminin in the extracellular matrix to promote P08253 expression and enhance ovarian cancer cell invasion . Modulation of diabetes with gonadotropin-releasing hormone antagonists in the nonobese mouse model of autoimmune diabetes . The nonobese mouse model of autoimmune diabetes ( NOD mouse ) exhibits a strain-dependent preponderance of disease in females . Castration of male NOD mice leads to an increased incidence of diabetes , suggesting that testosterone directly modulates the expression of diabetes in the NOD mouse . However , castration also modulates hypothalamic and pituitary hormone production via removal of the negative feedback effects of testosterone . One hypothalamic hormone with immunomodulatory properties whose expression is increased by castration is DB00644 . To test whether the increased incidence of diabetes in castrated male NOD mice is related to an increase in DB00644 activity , we treated castrated male NOD mice with Antide , a P30968 antagonist , to determine the effect on the incidence and timing of onset of diabetes . The prevalence of diabetes at 40 wk of age in male NOD mice was 50 % in sham-operated mice , compared with an 83 % prevalence in castrated males . Antide administration prevented the increased incidence of diabetes in the castrated male mice . Antide reduced total serum IgG levels , P05231 cytokine expression in cultured splenocytes , and the lymphocytic infiltration of islets . DB00644 administration exerted reciprocal effects , leading to earlier timing of onset of diabetes and increases in serum total IgG levels . We conclude that DB00644 modulates the expression of diabetes in the NOD mouse independently of gonadal steroids . Overexpression of folate binding protein alpha is one of the mechanism explaining the adaptation of HT29 cells to high concentration of methotrexate . The human colon adenocarcinoma cell line HT29 can be adapted to 10(-7)- 10(-4) M concentrations of methotrexate ( MTX ) . Cells adapted to 10(-4) M MTX have an enterocyte-like phenotype with P00374 gene amplification . Presently , we hypothetized that an increased expression of folate binding protein ( FBP ) may participate to the MTX resistance of 10(-4) MTX HT29 cells . The cDNA FBPalpha/beta-actin ratio of amplified transcripts was 4.8- and 1.5- fold higher in 10(-4) and in 10(-7) M MTX HT29 respectively , than in standard type HT29 cells . An increase of transcript level was observed when decreasing folic acid concentration . PI- P98160 cleaved 7.7 times more membrane FBP in 10(-4) M than in 10(-7) M MTX and wild type HT29 cells . In contrast to 10(-7) M MTX cells , growth of 10(-4) M MTX cells was dependent on folic acid concentration and abolished at a concentration lower than 0.9 microM . In conclusion , the adaptive mechanism of HT29 cells resistant to 10(-4) M MTX is the result of the synergistic overexpression of both P00374 and FBPalpha . Overexpression of FBPalpha may be related to the enterocyte-like phenotype of the cells . Production and characterization of antibodies to gonadotropin-releasing hormone receptor . Antibodies to the gonadotropin-releasing hormone ( DB00644 ) receptor of bovine pituitary membranes have been raised in rabbits by immunization with affinity-purified receptor preparations . These antibodies did not compete with 125I-labeled DB00644 analog ( DB06719 ) for binding to the receptors but did precipitate rat and bovine solubilized receptors labeled with 125I- DB06719 . Binding of the antibodies to the receptors was also demonstrated by immunoprecipitation of 125I-labeled purified receptors and photoaffinity-labeled receptors . The antibodies did not have a DB00644 -like activity but rather inhibited , in a dose-dependent manner , DB00644 -stimulated luteinizing hormone release from cultured rat pituitary cells . In addition , the antibodies did not inhibit luteinizing hormone release stimulated by high K+ concentration . This suggests that the antibodies recognize domains of the receptor other than the binding site of the hormone and thereby inhibit the biological response . These P30968 antibodies provide a useful tool for studying P30968 structure , function , localization , and biosynthesis . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . Salmon DB00644 and its analogues bind the human placental receptor . OBJECTIVE : The presence of DB00644 receptors in the human placenta has been recognized for a number of years . However , mammalian DB00644 , which is expressed in placental tissues , has limited affinity for the chorionic receptor . On the basis of immunological and bioactivity data , we have previously proposed that the chorionic DB00644 may differ from mammalian DB00644 . METHODS : We have studied the affinity of another isoform of DB00644 ( ie , salmon DB00644 and stable analogues of this DB00644 isoform ) , and compared their receptor affinity to that of mammalian DB00644 and its analogues . RESULTS : Using our receptor assay method with the labeled mammalian DB00644 analogue DB06719 , salmon DB00644 had a twofold greater affinity for the placental P30968 than did mammalian DB00644 and for the stable salmon DB00644 analogue the affinity was increased tenfold . Using a homologous receptor assay method with a stable salmon DB00644 analogue as label , the affinity for this salmon DB00644 analogue had a K(d) of 101 nmol/L . CONCLUSION : The presence of these higher affinity receptors for non-mammalian DB00644 in the human placenta has led us to propose that the chorionic tissues may express more than one isoform of DB00644 and that non-mammalian DB00644 , such as salmon DB00644 , may be potent regulators of placental functions . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . Accumulation of biglycan and perlecan , but not versican , in lesions of murine models of atherosclerosis . Proteoglycan accumulation within the arterial intima has been implicated in lipoprotein retention and in atherosclerosis progression in humans . Two commonly studied murine models of atherosclerosis , the apolipoprotein E ( apoE ) -deficient ( apoE-/- ) mouse and the low density lipoprotein receptor-deficient ( P01130 -/- ) mouse , develop arterial lesions similar to those of human atherosclerosis . However , specific proteoglycan classes that accumulate in lesions of these mice and their relation to the retention of specific apolipoproteins have not been previously determined . In this report , we characterized the distribution of proteoglycans ( versican , biglycan , and perlecan ) and apolipoproteins ( apoB , apoA-I , and apoE ) in proximal aortic lesions of chow-fed apoE-/- and P01130 -/- mice at 10 , 52 , and 73 weeks of age . We observed that similar to the apoE-/- mice , the P01130 -/- mice develop intermediate and advanced plaques within 52 weeks of age . P98160 and biglycan ( both are proteoglycans ) appeared early in lesion development with distinct expression patterns as the plaques advanced . Versican , a major proteoglycan detected in human plaques , was mostly absent in both strains . P02647 and apoB were detected in early through advanced lesions in regions of proteoglycan accumulation in both strains . Our results indicate that proteoglycans may contribute to the retention of lipoproteins at the earliest stage of atherosclerosis in murine models of atherosclerosis . Surfactin exhibits neuroprotective effects by inhibiting amyloid β-mediated microglial activation . Microglial-mediated neuroinflammation and neurotoxicity contribute to the pathogenesis of neurodegenerative diseases including Alzheimer 's disease ; therefore , control of microglial activation and subsequent suppression of neurotoxic pro-inflammatory molecules could provide a potential therapeutic approach for the treatment of such diseases . In this study , we investigated the effects of surfactin , a surfactant from Bacillus subtilis , on oligomeric amyloid β ( Aβ ) -induced microglial activation and neurotoxicity . Surfactin significantly suppressed expression of P14780 , P35228 and P35354 , as well as production of ROS , NO , DB00917 , P01375 -α , IL-1β , P05231 and P13500 in Aβ-stimulated BV-2 microglial cells . Moreover , surfactin markedly inhibited Aβ-induced nuclear translocation and activation of NF-κB as well as phosphorylation of JNK and p38 MAPK . Furthermore , surfactin protected hippocampal HT22 cells from indirect neuronal toxicity mediated by Aβ-treated microglial cells , but had no effect on Aβ-induced direct toxicity to HT22 cells . These results suggest that surfactin impairs the Aβ-induced inflammatory response of microglial cells and confers protection against indirect neurotoxicity to hippocampal cells . Our findings indicate that surfactin may have therapeutic potential for ameliorating Alzheimer 's disease as well as other neurodegenerative disorders which involve neuroinflammation . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Expression of type I P01148 receptor and in vivo and in vitro P01148 -I effects in corpora lutea of pseudopregnant rabbits . The expression of type I P01148 receptor ( P30968 -I ) and the direct role of P01148 -I on corpora lutea ( CL ) function were studied in the pseudopregnant rabbit model . Immunohistochemistry evidenced P30968 -I and P01148 -I in luteal cells at early ( day 4 pseudopregnancy ) - , mid ( day 9 ) - , and late ( day 13 ) -luteal stages . Real-time RT-PCR and western blotting revealed P30968 -I mRNA and protein at the three luteal stages . DB06719 in vivo treatment at days 9 and 13 decreased plasma progesterone levels for 48 and 24 h respectively . In in vitro cultured CL , buserelin reduced progesterone secretion , increased prostaglandin F(2α) ( P49763 (2α) ) secretion and cyclo-oxygenase-2 ( P35354 ) and nitric oxide synthase ( NOS ) activities at days 9 and 13 , and decreased PGE₂ at day 13 . Co-incubation with antagonists for P01148 -I ( antide ) , inositol 1,4,5-trisphosphate ( IP₃ , 2-amino-ethoxydiphenylborate ) , and diacylglycerol ( DAG , 1-hexadecyl-2-acetyl glycerol ) or inhibitors for phospholipase C ( P98160 , compound 48/80 ) , and protein kinase C ( PKC , staurosporine ) counteracted the buserelin effects . DB06719 co-incubated with P36551 inhibitor ( acetylsalicylic acid ) increased progesterone and decreased P49763 (2α) and NOS activity at days 9 and 13 , whereas co-incubation with NOS inhibitor ( DB04223 methyl ester ) increased progesterone at the same luteal stages . These results suggest that P30968 -I is constitutively expressed in rabbit CL independently of luteal stage , whereas P01148 -I down-regulates directly CL progesterone production via P49763 (2α) at mid- and late-luteal stages of pseudopregnancy , utilizing its cognate type I receptor with a post-receptorial mechanism that involves P98160 , IP₃ , DAG , PKC , P35354 , and NOS . Molecular phylogeny of Trypanosoma cruzi from Central America ( Guatemala ) and a comparison with South American strains . Molecular phylogenetic analysis was carried out for 21 strains of Trypanosoma cruzi , nine of which were obtained from Guatemala and 12 from South America . Phylogenetic trees were constructed using the nucleotide sequences of two nuclear gene regions , dihydrofolate reductase-thymidylate synthase ( P00374 -TS ) and trypanothione reductase ( TR ) , and contiguous portions of two mitochondrial genes , cytochrome oxidase subunit II ( P00403 ) and reduced nicotinamide adenine dinucleotide dehydrogenase subunit 1 ( P03886 ) . Possible genetic exchange between the rather divergent lineages of T. cruzi II from South America was suggested in the trees of the two nuclear genes . T. cruzi I strains obtained from Guatemala and Colombia were identical in all the genes examined , but other T. cruzi I isolates from South America were rather polymorphic in the P00374 -TS and mitochondrial genes . No genetic exchange was identified between T. cruzi I populations from Central and South America in the present study . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . Modulation of the P22301 /IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis . IFN-beta has been shown to be effective in the treatment of multiple sclerosis ( MS ) . However , the primary mechanism by which IFN-beta mediates its therapeutic effect remains unclear . Recent studies indicate that under defined conditions , IFN-beta may downregulate DC expression of IL-12 . We and others have shown that IFN-beta may also downregulate P22301 . In light of the recently proposed paradigm that an P22301 /IL-12 immunoregulatory circuit controls susceptibility to autoimmune disease , we examined the effect of IFN-beta on the development and behavior of the autoreactive T cell repertoire during experimental autoimmune encephalomyelitis ( EAE ) , an animal model sharing many features with MS . SWXJ mice were immunized with the immunodominant p139-151 determinant of myelin proteolipid protein ( PLP ) , and at onset of EAE were treated every other day with IFN-beta . After eight weeks of treatment , we assessed autoreactivity and observed no significant IFN-beta effect on splenocyte proliferation or splenocyte production of P01579 , P60568 , P05112 , or P05113 in response to the priming determinant used to initiate disease . However , in IFN-beta treated mice , the cytokine profile in response to the priming immunogen was significantly skewed toward an increased production of P22301 and a concurrent decreased production of IL-12 . Moreover , the in vivo modulation of the P22301 /IL-12 immunoregulatory circuit in response to the priming immunogen was accompanied by an aborted development of epitope spreading . Our results indicate that IFN-beta induces a reciprocal modulation of the P22301 /IL-12 cytokine circuit in vivo . This skewed autoreactivity establishes an inflammatory microenvironment that effectively prevents endogenous self-priming thereby inhibiting the progression of disease associated with epitope spreading . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Wheat germ agglutinin behaves as a DB00644 antagonist but induces gonadotrope desensitization . Preincubation of cultured rat pituitary cells with 10 micrograms/ml of either wheat germ agglutinin ( WGA ) or concanavalin A inhibited LH release stimulated with DB00644 ( 0.5 nM ) by 55 % and 40 % , respectively . WGA-inhibition of LH release stimulated by DB00644 was dose-dependent , reaching a plateau of 75 % inhibition at 50 micrograms/ml . Concomitantly , WGA induced a dose-dependent inhibition of 125I- DB06719 specific binding to pituitary cells , with a maximal inhibition of 45 % . The inhibition of 125I- DB06719 binding by WGA is due to P30968 internalization and not to persistent occupancy of the receptors . In addition to the effect of WGA on receptor internalization , WGA also induced partial desensitization of pituitary cells but was ineffective in modulating DB00644 -induced desensitization . These findings indicate that WGA has all the characteristics of a DB00644 antagonist , nevertheless , it does induce desensitization of cultured rat pituitary cells to further stimulation with DB00644 . Regulation of extracellular signal-regulated kinase 1/2 influences hippocampal neuronal survival in a rat model of diabetic cerebral ischemia . Activation of extracellular signal-regulated kinase 1/2 has been demonstrated in acute brain ischemia . We hypothesized that activated extracellular signal-regulated kinase 1/2 can protect hippocampal neurons from injury in a diabetic model after cerebral ischemia/reperfusion . In this study , transient whole-brain ischemia was induced by four-vessel occlusion in normal and diabetic rats , and extracellular signal-regulated kinase 1/2 inhibitor ( U0126 ) was administered into diabetic rats 30 minutes before ischemia as a pretreatment . Results showed that the number of surviving neurons in the hippocampal P00915 region was reduced , extracellular signal-regulated kinase 1/2 phosphorylation and P12956 activity were decreased , and pro-apoptotic Bax expression was upregulated after intervention using U0126 . These findings demonstrate that inhibition of extracellular signal-regulated kinase 1/2 activity aggravated neuronal loss in the hippocampus in a diabetic rat after cerebral ischemia/reperfusion , further decreased DNA repairing ability and accelerated apoptosis in hippocampal neurons . P27361 /2 activation plays a neuroprotective role in hippocampal neurons in a diabetic rat after cerebral ischemia/reperfusion . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . DB11320 contributes to tissue remodeling via periostin expression . DB11320 is thought to have a critical role in the synthesis of extracellular matrix in skin and may be involved in tissue remodeling of allergic diseases . Recent studies revealed that periostin , a matricelluar protein , contributed to tissue remodeling ; however , a link between periostin and histamine remains unproven . We investigated whether periostin was involved in histamine-induced collagen production . Cultured dermal fibroblasts derived from wild-type ( WT ) or periostin knockout ( PN(-/-) ) mice were stimulated with histamine , and then collagen and periostin production was evaluated . DB11320 induced collagen gene expression in WT fibroblasts in the late phase but not in the early phase , whereas no effect on collagen expression was observed in histamine-stimulated PN(-/-) fibroblasts . In WT fibroblasts , histamine directly induced periostin expression in a dose-dependent manner , and an H1 receptor antagonist blocked both periostin and collagen expression . DB11320 activated extracellular signal-regulated kinase 1/2 ( P27361 /2 ) through the H1 receptor . Q15063 induction was inhibited by either H1 antagonist or P27361 /2 inhibitor treatment in vitro and was attenuated in P35367 (-/-) mice . Elevated expression of periostin was found in lesional skin from atopic dermatitis patients . These results suggest that histamine mediates periostin induction and collagen production through activation of the H1 receptor-mediated P27361 /2 pathway ; furthermore , histamine may accelerate the chronicity of atopic dermatitis . Signaling and antiproliferative effects of type I and II gonadotropin-releasing hormone receptors in breast cancer cells . DB00644 receptors ( DB00644 -Rs ) mediate direct antiproliferative effects on hormone-dependent cancer cells . DB00644 -Rs can be grouped according to ligand specificity ( for DB00644 and -II ) , and there is evidence that type II DB00644 ligands and/or receptors can inhibit proliferation . Type I DB00644 -Rs ( e.g. human and sheep ) lack the C-terminal tails found in other G protein-coupled receptors including type II DB00644 -Rs ( e.g. Xenopus ; XGnRH-R ) . This underlies the remarkable resistance of type I DB00644 -Rs to desensitization and may be important for chronic effects on proliferation . To test this , we have compared the antiproliferative effects of DB00644 -Rs expressed in MCF7 breast cancer cells using recombinant adenovirus ( Ad ) . Endogenous DB00644 -Rs were not detected , but infection with Ad-expressing sheep DB00644 -Rs ( sGnRH-R ) facilitated proliferation inhibition by DB06719 , and maximum inhibition required only 10,000-20,000 sGnRH-Rs . XGnRH-Rs were much less efficient at inhibiting proliferation and were internalized faster than sGnRH-Rs . Thus , the type II P30968 is less efficient at inhibiting proliferation , presumably because it is rapidly desensitized and/or internalized . Moreover , comparisons of human P30968 , sGnRH-R , and XGnRH-R , as well as chimeric receptors ( type I DB00644 -Rs with C-terminal tails from XGnRH-Rs ) , revealed that C-terminal tail addition increases receptor expression and thereby increases the efficiency with which the vector facilitates the antiproliferative effect . The opposite effects of P40933 and Q9HBE4 on CLL B cells correlate with differential activation of the JAK/ P35610 and P27361 /2 pathways . The clonal expansion of chronic lymphocytic leukemia ( CLL ) cells requires the interaction with the microenvironment and is under the control of several cytokines . Here , we investigated the effect of P40933 and Q9HBE4 , which are closely related to P60568 and share the usage of the common gamma chain and of its P52333 -associated pathway . We found remarkable differences in the signal transduction pathways activated by these cytokines , which determined different responses in CLL cells . P40933 caused cell proliferation and prevented apoptosis induced by surface IgM cross-linking . These effects were more evident in cells stimulated via surface P25942 , which exhibited increased cell expression of IL-15Ralpha chain and , in some of the cases , also of IL-2Rbeta . Q9HBE4 failed to induce CLL cell proliferation and instead promoted apoptosis . Following cell exposure to P40933 , phosphorylation of P42229 was predominantly observed , whereas , following stimulation with Q9HBE4 , there was predominant P42224 and P40763 activation . Moreover , P40933 but not Q9HBE4 caused an increased phosphorylation of Shc and P27361 /2 . Pharmacological inhibition of P52333 or of MEK , which phosphorylates P27361 /2 , efficiently blocked P40933 -induced CLL cell proliferation and the antiapoptotic effect of this cytokine . The knowledge of the signaling pathways regulating CLL cell survival and proliferation may provide new molecular targets for therapeutic intervention . Purification and characterization of a high molecular weight histone deacetylase complex ( Q92769 ) of maize embryos . The dynamic state of core histone acetylation is maintained by histone acetyltransferases and deacetylases . In germinating maize embryos , four nuclear histone deacetylases can be distinguished . From a chromatin fraction prepared at 72 h after start of embryo germination , we have purified the nuclear histone deacetylase Q92769 to homogeneity . Using a sequence of chromatographic steps , we achieved the purification of an enzymatically active high molecular weight protein complex with an apparent molecular mass of 400 kDa , as determined by gel filtration chromatography . The purified enzyme was characterized in terms of enzymatic and kinetic properties , and sensitivity to several histone deacetylase inhibitors . In SDS-polyacrylamide gels , Q92769 split into three polypeptides of 45 , 42 , and 39 kDa , suggesting that the native enzyme is a multimer-protein complex . Electrophoresis under nondenaturing conditions in combination with second dimension SDS-gel electrophoresis indicated that all three protein components of the Q92769 complex were enzymatically active . Polyclonal antibodies against each of the three polypeptides were raised in rabbits . Each antiserum reacted with all three polypeptides on Western blots , suggesting that P29466 , Q8NFH3 , and p39 are highly homologous . This homology was confirmed by amino acid sequencing of peptides generated from each of the three Q92769 components . [ The effects of DB00644 agonist on steroidogenesis in the rat ovary ] . The effects of DB00644 agonist ( buserelin ) on in vitro ovarian steroidogenesis were studied using DES-treated immature rats and PMS-treated immature rats . The estradiol and progesterone secreted by the cultured ovarian cells and the activities of various enzymes of steroid-metabolism were examined with or without gonadotropins ( DB00094 or hCG ) , and the effects of 10(-6)-10(-12) M of buserelin on those indices were observed for 3-72 hours . In addition , the kinetic study of ovarian P30968 was performed using 125I-labelled buserelin and crude ovarian cell membrane fraction of PMS-treated rats . The Scatchard analysis revealed the specific high affinity and low capacity ovarian P30968 ( Kd = 0.92 nM and Bmax = 0.57 fmol/mg tissue ) . The DB00094 -stimulated cholesterol side chain cleavage enzyme ( CSCC ) activity of the DES-treated rats was suppressed in a dose-dependent manner by buserelin . Estradiol release and aromatase activity were increased by 10(-8) M buserelin within 48 hours from the beginning of the incubation of the PMS-treated rat ovarian cells , but were suppressed after 48 hours . DB06719 increased basal progesterone secretion and both activities of CSCC and of 3 beta-hydroxysteroid dehydrogenase of PMS-treated rat ovarian cells incubated without hCG , which were suppressed by buserelin co-incubated with 100 IU/ml of hCG . These results suggested that DB00644 plays a physiological role in ovarian steroidogenesis binding the specific receptor and that DB00644 promotes the development of the follicle through increased estrogen synthesis in the early stage of the folliculogenesis and the luteinization in the late stage of the follicular development through increased progesterone and decreased estradiol production and the luteolysis in the luteinized cells by hCG through decreased progesterone secretion . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . [ 5-hydroxytryptamine ( serotonin ) receptors -- nomenclature and classification of types and subtypes ] . 5-HT receptors represent a superfamily of receptors with the largest known number of receptor subtypes . At present 15 receptor subtypes of three groups has been recognized . The 5-HT1 subfamily of receptors contains subtypes P08908 , P28222 , P28221 , P28566 , and P30939 ; activation of all of them results in the inhibition of adenylylcyclase . The subfamily of 5-HT2 contains subtypes 5- Q13049 , P41595 , and P28335 ; their activation leads to the stimulation of P98160 . Finally , subfamily of miscellaneous 5-HT receptors contains subtypes 5- Q9H205 , Q13639 , 5-HT5 , P50406 , and P34969 ; some of them has been cloned , however , our knowledge on their function is still minimal . 5-HT receptors participate in many physiological functions and a disturbance in serotonergic neurotransmission might cause several types of disease . 5-HT plays an important role in depression ; to cure this disease , drugs which increase levels of this neurotransmitter are used . A new drug group called Selective Serotonin Reuptake Inhibitors ( SSRI ) has been recently discovered . These drugs block the reuptake of 5-HT into nerve endings . There is an intensive search for new selective agonists as well as antagonists which could be use not only in the classification of receptor subtypes but which also possess certain therapeutic potential . Induction of O15534 mRNA expression in immortalized gonadotropes by gonadotropin-releasing hormone ( DB00644 ) : involvement of protein kinase C and Q96HU1 kinase signaling . The initiation and maintenance of reproductive function in mammals is critically dependent on the pulsatile secretion of gonadotropin-releasing hormone ( DB00644 ) . This peptide drives the pulsatile release of DB00094 and LH from the pituitary pars distalis via signaling pathways that are activated by the type I P30968 ( P30968 ) . Recently , a microarray analysis study reported that a number of genes , including mPer1 , are induced by DB00644 in immortalized gonadotrope cells . In view of these data , we have begun to analyze in detail the signaling pathways mediating the action of DB00644 on mPer1 expression in these cells . Using quantitative real-time polymprose cho read ( PCR ) , we could confirm that exposure of immortalized gonadotropes ( LbetaT2 cells ) to the DB00644 analog , buserelin , markedly induces mPer1 ( but not mPer2 ) expression . Consistent with P30968 signaling via the protein kinase ( PK ) -C pathway , exposure of the cells to phorbol 12,13-dibutyrate rapidly elevates both mPer1 and LHbeta subunit mRNA levels , while pharmacological inhibition of PKC prevents the mPer1 and LHbeta response to buserelin . As DB00644 is known to regulate gonadotropin synthesis via activation of Q8NFH3 /44 mitogen-activated protein kinase ( MAPK ) signaling pathways , we then examined the involvement of this pathway in regulating mPer1 expression in gonadotropes . Our data reveal that DB00644 -induced mPer1 expression is blocked following acute exposure to a MAPK kinase inhibitor . Although the involvement of this signaling mechanism in the regulation of mPer1 is known in neurons , e.g. , in the suprachiasmatic nuclei , the induction of mPer1 in gonadotropes represents a novel mechanism of DB00644 signaling , whose functional significance is still under investigation . Amyloid beta-derived neuroplasticity in bone marrow-derived DB05914 is mediated by P01303 and P41595 receptors via P27361 /2 signalling pathways . OBJECTIVE : In Alzheimer 's disease , toxic soluble and insoluble forms of amyloid beta ( Abeta ) cause synaptic dysfunction and neuronal loss . Given its potential role in producing a toxic host microenvironment for transplanted donor stem cells , we investigated the interaction between Abeta and proliferation , survival , and differentiation of bone marrow-derived DB05914 ( BM- O60682 ) in culture . MATERIALS AND METHODS : We used BM- O60682 that had been isolated from mouse bone marrow and cultured , and we also assessed relevant reaction mechanisms using gene microarray , immunocytochemistry , and inhibitors of potential signalling molecules , such as mitogen-activated protein kinase ( MAPK ) /extracellular signal-regulated kinase ( P29323 )1/2 and tyrosine protein kinase . RESULTS AND CONCLUSIONS : Interestingly , we found that treatment with aggregated ( 1-40 or 1-42 ) and oligomeric ( 1-42 ) Abeta promoted neuronal-like differentiation of BM- O60682 without toxic effects . This was not dependent on soluble factors released from BM- O60682 progeny nor solely on formation of Abeta fibrils . The effect of Abeta is mediated by G-protein coupled receptors , neuropeptide Q03519 ( P25929 ) and serotonin ( 5-hydroxytryptamine ) receptor 2B , via phosphatidylinositol-3-OH kinase-dependent activation of the MAPK/ P27361 /2 . Our results lend support to the idea that reciprocal donor stem cell-host interactions may promote a regenerative response that can be exploited by epigenetic modulation of P01303 /serotonergic gene expression , for stem cell therapy , in Alzheimer 's disease . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Transient transfection of GGH3-1 ' cells [ GH3 cells stably transfected with the gonadotropin-releasing hormone ( DB00644 ) receptor complementary deoxyribonucleic acid ] with the carboxyl-terminal of beta-adrenergic receptor kinase 1 blocks prolactin release : evidence for a role of the G protein beta gamma-subunit complex in DB00644 signal transduction . G proteins consist of heterotrimeric alpha- , beta- , and gamma-subunits . To assess the role of the beta gamma-subunit complex in P30968 -mediated signal transduction , GGH3-1 ' cells were transfected with plasmids PRK5-beta O14965 ( 495-689 ) containing complementary DNA ( cDNA ) of the carboxyl-terminal ( Gly495-Leu689 ) of beta-adrenergic receptor kinase 1 ( beta O14965 ) . GGH3-1 ' cells are GH3 cells that have been stably transfected with rat P30968 cDNA . The carboxyl region of beta O14965 ( Gly495-Leu689 ) binds to the beta gamma complex and thereby inhibits its action . Twenty-four hours after stimulation , PRL release , DB02527 release , and inositol phosphate ( IP ) production were measured in these cells and in control cells transfected with vector PRK5 cDNA alone . In cells expressing the beta O14965 -(495-689) sequence there was inhibition of basal PRL release ( 69.3 % ) , DB02527 release ( 61.2 % ) , and IP production ( 75.5 % ) compared to cells containing vector only . When cells expressing the beta O14965 fragment were stimulated with a DB00644 analog ( DB06719 ; 10(-7) M ) , maximal responses were inhibited ( 66.1 % for PRL release , 52.3 % for DB02527 release , and 79.1 % for IP production ) . Scatchard analysis of DB00644 analog binding was also performed in the two groups of transfected cells . No significant differences in Kd or receptor numbers were found between beta O14965 -(495-689)-transfected cells and control cells containing the vector alone . These data suggest a role for the beta gamma complex in mediation of DB02527 release , IP production , and hormone release in response to agonist occupancy of the P30968 . In vivo effects of a combined P28222 receptor/ P31645 antagonist in experimental pulmonary hypertension . AIMS : A mechanism for co-operation between the serotonin ( 5-hydroxytryptamine , 5-HT ) transporter and P28222 receptor in mediating pulmonary artery vasoconstriction and proliferation of pulmonary artery smooth muscle cells has been demonstrated in vitro . Here we determine , for the first time , the in vivo effects of a combined P28222 receptor/serotonin transporter antagonist ( LY393558 ) with respect to the development of pulmonary arterial hypertension ( PAH ) and its in vitro effects in human pulmonary artery smooth muscle cells ( hPASMCs ) derived from idiopathic PAH ( IPAH ) patients . METHODS AND RESULTS : We determined the effects of LY393558 as well as a selective serotonin transporter inhibitor , citalopram , on right ventricular pressure , right ventricular hypertrophy , and pulmonary vascular remodelling in wildtype mice and mice over-expressing serotonin transporter ( P31645 + mice ) before and after hypoxic exposure . We also compared their effectiveness at reversing PAH in P31645 + mice and hypoxic mice . Further , we examined the proliferative response to serotonin in IPAH hPASMCs . We also clarified the pharmacology of serotonin-induced vasoconstriction and P28222 receptor/serotonin transporter interactions in mouse isolated pulmonary artery . DB00215 had a moderate effect at preventing and reversing experimental PAH in vivo whereas LY393558 was more effective . LY393558 was more effective than citalopram at reversing serotonin-induced proliferation in IPAH hPASMCs . There is synergy between P28222 receptor and serotonin transporter inhibitors against serotonin-induced vasoconstriction in mouse pulmonary arteries . CONCLUSION : P28222 receptor and serotonin transporter inhibition are effective at preventing and reversing experimental PAH and serotonin-induced proliferation of PASMCs derived from IPAH patients . Targeting both the serotonin transporter and P28222 receptor may be a novel therapeutic approach to PAH . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Cyclic adenosine 3',5'-monophosphate ( DB02527 ) and DB02527 responsive element-binding protein are involved in the transcriptional regulation of gonadotropin-releasing hormone ( DB00644 ) receptor by DB00644 and mitogen-activated protein kinase signal transduction pathway in Q92820 (3) cells . Stimulation of mouse P30968 promoter by a DB00644 agonist ( DB06719 ) , or by a DB02527 analogue , significantly increased reporter ( luciferase ) activity . Overexpression of P04049 , P27361 , or P28482 partially blocked DB06719 -stimulated luciferase activity . In contrast , treatment with a mitogen-activated protein kinase ( MAPK ) kinase inhibitor ( PD 98059 ) activated basal and DB06719 -stimulated luciferase activity in a dose-dependent manner . Transient transfection of the deleted DB02527 response element expression vector followed by pretreatment with PD98059 prior to DB06719 stimulation showed that the transcriptional response was decreased compared to wild-type promoter . A gel-mobility shift assay using a probe containing the DB02527 response element showed the presence of two specific protein-DNA complexes that contain one or more members of the DB02527 responsive element-binding ( CREB ) protein family . These results suggest that DB02527 and CREB participate in the DB00644 activation of P30968 promoter activity and that the MAPK cascade is involved in the negative regulation of basal and DB00644 -stimulated P30968 transcriptional activity . Heart allograft protection with low-dose carbon monoxide inhalation : effects on inflammatory mediators and alloreactive T-cell responses . BACKGROUND : DB11588 ( CO ) , a byproduct of heme catalysis , has lately received considerable attention as a regulatory molecule in cellular and biological processes . CO has been shown to provide potent protection against a variety of tissue injuries . We hypothesized in this study that low concentration CO would be beneficial for organ allografts , which frequently undergo several types of injury such as ischemia/reperfusion , alloimmune reaction , and inflammation METHODS : The efficacy of low-dose CO was examined in a fully allogeneic LEW to BN rat heterotopic heart transplantation ( HHTx ) model . Recipients were kept in air or exposed to low-dose CO ( 20 ppm ) for 14 , 28 , or 100 days after HHTx under short-course tacrolimus RESULTS : CO treatment ( d0-28 , 0-100 ) was remarkably effective in prolonging heart allograft survival to a median of > 100 from 45 days in the air-control group , with significant reductions of arteritis , fibrosis , and cellular infiltration , including macrophages and T cells . CO inhibited intragraft upregulation of Th1 type cytokines ( P60568 , IFNgamma ) , proinflammatory mediators ( IL-1beta , TNFalpha , P05231 , P35354 ) , and adhesion molecule . Shorter CO exposure in early ( 0-13d ) and late ( 14-28d ) posttransplant periods also prolonged graft survival , with a significant inhibition of inflammatory mediators CONCLUSIONS : These results show that low dose CO inhalation protects heart allografts and can considerably prolong their survival . CO appears to function via multiple mechanisms , including direct inhibition of Th1 type cytokine production and regulation of inflammatory responses . In vitro effects of gonadotropin-releasing hormone ( DB00644 ) analogue on cancer cell sensitivity to cis-platinum . Six endometrial cancer cell lines ( Ishikawa , EIIL , HEC1A , 6 , 50 and 59 ) , one breast cancer cell line ( MCF-7 ) and two ovarian cancer cell lines ( OVHS-1 , HRA ) were treated for 24 or 168 h with a gonadotropin-releasing hormone ( DB00644 ) analogue , DB06719 acetate , and the cellular growth profile was studied . All these cell lines except for the HRA line had positive P30968 mRNA expression detected by reverse transcriptase polymerase chain reaction . GnRHa suppressed cell growth after 168 h of exposure , but not after 24 h . Suppression of cell growth by the exposure to cis-platinum ( DB00515 , 10 nM for 24 h ) was significantly increased in the presence of GnRHa for 168 h . The mechanism of this growth inhibition was tested by examining both RNA components of human telomerase ( hTR ) expression and telomerase activity . The results showed that GnRHa inhibits telomerase activity without altering the RNA component of telomerase expression . The present data suggest that DB00644 analogue may modulate endometrial , breast and ovarian cancer cell growth through modifying the telomerase activity . Since GnRHa increased the cytotoxic effects of DB00515 and GnRHa is a compound of high patient compliance , the value of GnRHa as a tumor sensitizer to DB00515 should be further tested in clinical trials . [ Anti-cholesterol agents , new therapeutic approaches ] . Statins and fibrates constitute the two major families of lipid-lowering agents . Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia . Both drugs are also used for the treatment of mixed dyslipidemia . Some fibrates efficiently lower serum LDL-cholesterol . Statins inhibit P04035 and decrease cellular cholesterol synthesis . The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the P01130 gene . This over expression of the P01130 in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels . The effects of fibrates on lipid metabolism are entirely due to their capacity to activate Q07869 and to induce the over expression of genes containing a PPRE in their promoter . Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis . Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription , respectively . Fibrates could decrease triglycerides partly by inducing apo A-V over-expression . These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription . Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . Transcriptional regulation of the gonadotropin-releasing hormone receptor gene is mediated in part by a putative repressor element and by the cyclic adenosine 3',5'-monophosphate response element . The levels of the P30968 ( GnRHR ) and its messenger RNA depend on the pattern of administration of DB00644 . In this study , internal deletion mutants in a luciferase reporter gene vector ( GnRHR-pXP2 ) containing a 1226-bp promoter fragment of mouse GnRHR gene were used to examine the regulation of GnRHR gene transcription in GGH3 cells . Our results indicate that the mouse GnRHR promoter contains one putative repressor element located at position -343/-335 . When this sequence was deleted , the GnRHR promoter activity was significantly increased in both basal and DB00644 agonist ( DB06719 ) - , phorbol ester- , and forskolin-stimulated cells . Gel mobility shift assay showed that the sequence -343/-335 is capable of binding GGH3 nuclear proteins . With deletion of the DB02527 response element ( -107/-100 ) , basal and DB06719 -stimulated transcription was decreased . The same response was observed after stimulation with forskolin . Stimulation with (Bu)2cAMP did not alter transcription above basal levels . The stimulation with phorbol ester resulted in an attenuated increase in transcriptional activity , suggesting that this sequence of the GnRHR promoter is a DB02527 response element . These results suggest that the transcriptional activity of the GnRHR gene is mediated in part by a putative repressor element and by the DB02527 response element . Biphasic action of cyclic adenosine 3',5'- monophosphate in gonadotropin-releasing hormone ( DB00644 ) analog-stimulated hormone release from GH3 cells stably transfected with P30968 complementary deoxyribonucleic acid . GH3 cells are a PRL-secreting adenoma cell line derived from pituitary lactotropes . These cells have been stably transfected with rat P30968 complementary DNA to produce four cell lines : Q92820 (3)1 ' , Q92820 (3)2 ' , Q92820 (3)6 ' , and Q92820 (3)12 ' . In response to either DB00644 or DB06719 ( a metabolically stable DB00644 agonist ) , these cell lines synthesize PRL in a DB02527 -dependent manner . Only Q92820 (3)6 ' cells desensitize in response to persistent treatment with 10(-7) g/ml DB06719 . Q92820 (3)1 ' , Q92820 (3)2 ' , and Q92820 (3)12 ' cells , however , can be made refractory to DB06719 stimulation by raising DB02527 levels either by the addition of (Bu)2cAMP to the medium or by treatment with cholera toxin . In Q92820 (3) cells , low levels of DB02527 fulfill the requirements for a second messenger , whereas higher levels appear to mediate the development of desensitization . The observation that in Q92820 (3)6 ' cells , DB02527 production persists after the onset of desensitization is consistent with the view that the mechanism responsible for desensitization is distal to the production of DB02527 . Moreover , the absence of any significant difference in the amount of DB02527 produced per cell in Q92820 (3)2 ' , Q92820 (3)6 ' , or Q92820 (3)12 ' cells suggests that elevated DB02527 production per cell does not explain the development of desensitization in Q92820 (3)6 ' cells . We suggest that DB06719 -stimulated PRL synthesis in Q92820 (3)6 ' cells is mediated by a different DB02527 -dependent protein kinase pool(s) than that in nondesensitizing Q92820 (3) cells . Such a protein kinase A pool(s) may be more susceptible to degradation via DB02527 -mediated mechanisms than the protein kinase pools mediating the DB06719 response in nondesensitizing Q92820 (3) cells . A similar mechanism has been reported in other systems . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . Low density lipoprotein receptor-related protein ( Q14764 ) is required for lactoferrin-enhanced collagen gel contractile activity of human fibroblasts . Fibroblasts plated on a type I collagen gel can reduce the size of the gel in a way that mimics the reorganization of the collagen matrix that accompanies the wound healing process . We demonstrated previously that lactoferrin ( Lf ) specifically binds to WI-38 human fibroblasts and enhances their collagen gel contractile activity . The effect of Lf correlated with the phosphorylation of myosin light chain ( MLC ) , suggesting that Lf promotes fibroblast contractile activity by regulating MLC phosphorylation . We found here that the binding of Lf to WI-38 cells was inhibited by recombinant receptor-associated protein ( P30533 ) , a universal competitor for ligand binding to Q14764 ( P01130 -related protein ) , and P30533 can also promote the collagen gel contractile activity . These observations suggest that Q14764 is a receptor that mediates the Lf-induced enhancement of collagen gel contractile activity in WI-38 fibroblasts . To confirm the hypothesis , we utilized Q14764 antisense oligonucleotide , which was modified by morpholino linkage . Suppression of Q14764 expression abrogated the Lf-induced enhancement the contractile activity in fibroblasts . Treatment of fibroblasts with Lf enhanced the phosphorylation of P27361 /2 and the activation of MLC kinase ( MLCK ) . These effects were attenuated by suppression of Q14764 expression . These findings suggest that Q14764 is involved in the Lf-enhanced collagen gel contractile activity of WI-38 fibroblasts by converting the Lf binding signal into the activation of P27361 /2 and MLCK . Effects of gonadotropin-releasing hormone agonist on steroidogenesis in the rat ovary . To assess the regulatory roles of gonadotropin-releasing hormone ( DB00644 ) in ovarian function , the kinetics of the ovarian P30968 and the effects of the DB00644 superagonist buserelin on steroidogenesis in ovarian cell culture were examined . Scatchard analysis of buserelin-binding to crude ovarian cell membrane revealed a specific high-affinity P30968 . DB06719 together with follicle-stimulating hormone stimulated estradiol ( E2 ) production in immature follicles in hypophysectomized and DES-treated rats . On the other hand , applied to developing follicles of rats treated with pregnant-mare-serum gonadotropin buserelin suppressed E2 production to terminate follicle maturation and simultaneously stimulated progesterone ( P4 ) production to induce luteinization . With ovarian cells luteinized by human chorionic gonadotropin in vitro , buserelin suppressed production of both P4 and E2 , leading to luteolysis . DB06719 affected steroid production by modulating activities of key enzymes in steroid synthesis . These findings indicate that buserelin action depended on the gonadotropin priming of ovarian cells , and suggest the possible involvement of DB00644 in the regulation of steroidogenesis throughout the ovulatory cycle . Hormonal regulation of human trophoblast differentiation : a possible role for 17beta-estradiol and DB00644 . We have examined the role of 17beta-estradiol and gonadotropin releasing hormone ( DB00644 ) in the regulation of functional differentiation in human trophoblasts . In contrast to its recognized functions as a proliferation-promoting hormone in a variety of cell types , we found that 17beta-estradiol induced terminal differentiation in human trophoblastic cells , and that this event was estrogen-receptor-mediated . This process involved a loss in expression of Cyclins A2 and E , and a coincident increase in p27(Kip1) . The anti-proliferative effects of 17beta-estradiol were annulled by specific transforming growth factor-beta 1 ( TGFbeta1 ) -neutralizing antibody , suggesting that 17beta-estradiol may mediate its growth-inhibitory actions , through TGFbeta1 activity . Following exposure to DB06719 , cultured human trophoblastic cells stopped proliferating and formed functionally mature syncytiotrophoblasts . This differentiation event , that involved a drastic loss in expression of proliferating-cell-nuclear-antigen , could be blocked by DB00050 , suggesting the involvement of functional DB00644 receptors . Preliminary studies on the characterization of the human placental P30968 , indicate the presence of multiple receptor isoforms across human gestation . 25-Hydroxycholesterol is not a ligand for the orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) . The orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) is involved in the transcriptional regulation of all the steroid hydroxylase genes , and also regulates the transcription of the genes for Müllerian Inhibitory substance ( P03971 ) , alpha subunit of glycoprotein hormone , LHbeta , oxytocin , P30968 , Q01718 , prolactin receptor , DAX-1 , and steroidogenic acute regulatory protein . Other members of the nuclear receptor gene family , including steroid hormone , thyroid hormone , retinoic acid , Q07869 , and vitamin D receptors must bind ligand to activate transcription , but Q13285 has been considered to be an orphan nuclear receptor because , when identified , it had no known ligand . A recent publication suggested that transcriptional regulation by Q13285 , expressed in a non-steroidogenic CV-1 cells , could be activated by oxysterols suggesting that these compounds could serve as natural ligands for Q13285 . We now demonstrate that 25-hydroxycholesterol , either added exogenously or synthesized endogenously in steroidogenic mouse Leydig MA-10 cells , did not act as a ligand for Q13285 , as it did not increase transcription from six different Q13285 -dependent DNA sequences . Furthermore , the abundance of these oxysterols in MA-10 cells was much less than concentrations needed for activation of Q13285 in CV-1 cells , indicating that Q13285 is not constitutively bound by ligand in MA-10 cells . Thus , in steroidogenic cells , transcriptional regulation of the steroid hydroxylase genes by Q13285 does not depend upon the presence of 25-hydroxycholesterol , and is not modified by its presence . Primary cilia in stem cells and neural progenitors are regulated by neutral sphingomyelinase 2 and ceramide . We show here that human embryonic stem ( ES ) and induced pluripotent stem cell-derived neuroprogenitors ( NPs ) develop primary cilia . Ciliogenesis depends on the sphingolipid ceramide and its interaction with atypical PKC ( aPKC ) , both of which distribute to the primary cilium and the apicolateral cell membrane in NP rosettes . Neural differentiation of human ES cells to NPs is concurrent with a threefold elevation of ceramide-in particular , saturated , long-chain C16:0 ceramide ( N-palmitoyl sphingosine ) and nonsaturated , very long chain C24:1 ceramide ( N-nervonoyl sphingosine ) . Decreasing ceramide levels by inhibiting ceramide synthase or neutral sphingomyelinase 2 leads to translocation of membrane-bound aPKC to the cytosol , concurrent with its activation and the phosphorylation of its substrate O14965 ( AurA ) . Inhibition of aPKC , AurA , or a downstream target of AurA , Q9UBN7 , restores ciliogenesis in ceramide-depleted cells . Of importance , addition of exogenous C24:1 ceramide reestablishes membrane association of aPKC , restores primary cilia , and accelerates neural process formation . Taken together , these results suggest that ceramide prevents activation of Q9UBN7 by cytosolic aPKC and AurA , which promotes acetylation of tubulin in primary cilia and , potentially , neural processes . This is the first report on the critical role of ceramide generated by Q9NY59 in stem cell ciliogenesis and differentiation . RNA editing and mitochondrial activity in promastigotes and amastigotes of Leishmania donovani . Kinetoplast maxicircle DNA sequence organisation was investigated in Leishmania donovani , strain 1S LdBob . Gene arrangement in the coding ( conserved ) region of the maxicircle is collinear with that of most trypanosomatids , with individual genes showing 80-90 % nucleotide identity to Leishmania tarentolae , strain UC . The notable exception was an integration of a full-size minicircle sequence in the P03886 gene coding region found in L. donovani . Editing patterns of the mitochondrial mRNAs investigated also followed L. tarentolae UC patterns , including productive editing of the components of respiratory complexes III-V , and ribosomal protein P28222 ( P25398 ) , as well as the lack of productive editing in five out of six pan-edited cryptogenes ( P03897 , ND8 , ND9 , P46379 , G4 ) found in these species . Several guide RNAs for the editing events were localised in minicircles and maxicircles in the locations that are conserved between the species . Mitochondrial activity , including rates of oxygen consumption , the presence and the levels of respiratory complexes and their individual subunits and the steady-state levels of several mitochondrial-encoded mRNAs were essentially the same in axenically grown amastigotes and in promastigotes of L. donovani . However , some modulation of mitochondrial activity between these developmental stages was suggested by the finding of an amastigote-specific component in complex IV , a down-regulation of mitochondrial RNA-binding proteins ( MRP ) and MRP-associated protein ( MRP-AP ) in amastigotes , and by variations in the levels of P25398 , P03897 , ND9 , P46379 and G4 pre-edited transcripts . Tunicamycin and neuraminidase effects on luteinizing hormone ( LH ) -releasing hormone binding and LH release from rat pituitary cells in culture . We have studied the effects of tunicamycin ( TM ) and neuraminidase on the binding of 125I-labeled DB06719 , a DB00644 agonist , and on DB00644 -stimulated LH release in cultured rat pituitary cells . Treatment with TM , an antibiotic which inhibits protein glycosylation , abolished the development of elongated cell processes without any effect on cell viability . Concomitantly , TM caused a time- and dose-dependent inhibition of specific binding of DB06719 and of DB00644 -stimulated LH release . The inhibition of binding was due to a decrease in the number of DB00644 receptors without any significant effect on binding affinity . Protein synthesis was not affected under these experimental conditions , suggesting that the aglycosylated DB00644 receptors are probably intracellularly accumulated and are not expressed on the cell surface . Treatment with neuraminidase inhibited only 50 % of DB00644 agonist binding and did not affect DB00644 -stimulated LH release . These results indicate that the oligosaccharide portion is essential for the functional properties of the P30968 .

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interacts_with DB08901?

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Progressive changes in the leukemogenic signaling in P11274 / P00519 -transformed cells . Our previous study indicated that P11274 / P00519 SH2 domain and P11274 / P00519 SH3 domain+SH2 domain complex are required for immediate activation of the phosphatidylinositol-3 kinase PI-3k ) --> Akt serine/threonine kinase pathway and of the signal transducer and activator of transcription 5 ( P42229 ) , respectively , in hematopoietic cells . We show here that the defect in activation of PI-3k/Akt by P11274 / P00519 DeltaSH2 mutant ( SH2 domain deleted ) and of P42229 by P11274 / P00519 DeltaSH3+DeltaSH2 mutant ( SH3 and SH2 domains deleted ) is not permanent and both Akt and P42229 could be ' re-activated ' by in vitro culture . This phenomenon was responsible for increased resistance to apoptosis , growth factor-independent proliferation and leukemogenesis in SCID mice . Incubation of cells with P11274 / P00519 tyrosine kinase inhibitor STI571 abrogated the ' re-activation ' of Akt or P42229 by P11274 / P00519 SH3+SH2 mutants in some clones , in the others Akt and P42229 activation became independent on P11274 / P00519 kinase activity . The immediate upstream activators of Akt and P42229 such as PI-3k and Jak-2 were also activated . In addition , the common beta subunit of P08700 / P05113 /GM- P04141 receptor was tyrosine phosphorylated in the clones in which ' re-activation ' was dependent on the P11274 / P00519 kinase activity . These results suggested that ' re-activation ' of Akt and P42229 , in the absence of functional P11274 / P00519 SH3+SH2 domains , may be achieved by two different mechanisms : ( i ) P11274 / P00519 kinase-dependent activation of alternative pathway(s) and ( ii ) additional genetic changes stimulating Akt and P42229 independently of P11274 / P00519 . Oncogene ( 2000 ) 19 , 4117 - 4124 Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . DB08901 is a potent inhibitor of wild-type and drug-resistant gatekeeper mutant P07949 kinase . P07949 kinase is aberrantly activated in thyroid cancers and in rare cases of lung and colon cancer , and has been validated as a molecular target in these tumors . DB05294 was recently approved for the treatment of medullary thyroid cancer . However , vandetanib is ineffective in vitro against P07949 mutants carrying bulky aminoacids at position 804 , the gatekeeper residue , similarly to drug-resistant P11274 - P00519 mutants in chronic myeloid leukemia . DB08901 is a multi-target kinase inhibitor that was recently approved for treatment-refractory Philadelphia-positive leukemia . We show here potent inhibition of oncogenic P07949 by ponatinib , including the drug-insensitive V804M/L mutants . DB08901 inhibited the growth of P07949 + and P11274 - P00519 + cells with similar potency , while not affecting P07949 -negative cells . Both in biochemical and in cellular assays ponatinib compared favorably with known P07949 inhibitors , such as vandetanib , cabozantinib , sorafenib , sunitinib and motesanib , used as reference compounds . We suggest that ponatinib should be considered for the treatment of P07949 + tumors , in particular those expressing vandetanib-resistant V804M/L mutations . DB08901 is active against imatinib-resistant mutants of Q6UN15 - P16234 and P10721 , and against P11362 -derived fusion kinases . Mutation D816V alters the internal structure and dynamics of c- P10721 receptor cytoplasmic region : implications for dimerization and activation mechanisms . The type III receptor tyrosine kinase ( RTK ) P10721 plays a crucial role in the transmission of cellular signals through phosphorylation events that are associated with a switching of the protein conformation between inactive and active states . D816V P10721 mutation is associated with various pathologies including mastocytosis and cancers . D816V-mutated P10721 is constitutively active , and resistant to treatment with the anti-cancer drug Imatinib . To elucidate the activating molecular mechanism of this mutation , we applied a multi-approach procedure combining molecular dynamics ( MD ) simulations , normal modes analysis ( Q13145 ) and binding site prediction . Multiple 50-ns MD simulations of wild-type P10721 and its mutant D816V were recorded using the inactive auto-inhibited structure of the protein , characteristic of type III RTKs . Computed free energy differences enabled us to quantify the impact of D816V on protein stability in the inactive state . We evidenced a local structural alteration of the activation loop ( A-loop ) upon mutation , and a long-range structural re-organization of the juxta-membrane region ( JMR ) followed by a weakening of the interaction network with the kinase domain . A thorough normal mode analysis of several MD conformations led to a plausible molecular rationale to propose that JMR is able to depart its auto-inhibitory position more easily in the mutant than in wild-type P10721 and is thus able to promote kinase mutant dimerization without the need for extra-cellular ligand binding . Pocket detection at the surface of Q13145 -displaced conformations finally revealed that detachment of JMR from the kinase domain in the mutant was sufficient to open an access to the catalytic and substrate binding sites . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . P11274 - P00519 compound mutations combining key kinase domain positions confer clinical resistance to ponatinib in Ph chromosome-positive leukemia . DB08901 is the only currently approved tyrosine kinase inhibitor ( TKI ) that suppresses all P11274 - P00519 single mutants in Philadelphia chromosome-positive ( Ph(+) ) leukemia , including the recalcitrant P11274 - P00519 (T315I) mutant . However , emergence of compound mutations in a P11274 - P00519 allele may confer ponatinib resistance . We found that clinically reported P11274 - P00519 compound mutants center on 12 key positions and confer varying resistance to imatinib , nilotinib , dasatinib , ponatinib , rebastinib , and bosutinib . T315I-inclusive compound mutants confer high-level resistance to TKIs , including ponatinib . In vitro resistance profiling was predictive of treatment outcomes in Ph(+) leukemia patients . Structural explanations for compound mutation-based resistance were obtained through molecular dynamics simulations . Our findings demonstrate that P11274 - P00519 compound mutants confer different levels of TKI resistance , necessitating rational treatment selection to optimize clinical outcome . Potent activity of ponatinib ( DB08901 ) in models of P36888 -driven acute myeloid leukemia and other hematologic malignancies . DB08901 ( DB08901 ) is a novel multitargeted kinase inhibitor that potently inhibits native and mutant P11274 - P00519 at clinically achievable drug levels . DB08901 also has in vitro inhibitory activity against a discrete set of kinases implicated in the pathogenesis of other hematologic malignancies , including P36888 , P10721 , fibroblast growth factor receptor 1 ( P11362 ) , and platelet derived growth factor receptor α ( P09619 α ) . Here , using leukemic cell lines containing activated forms of each of these receptors , we show that ponatinib potently inhibits receptor phosphorylation and cellular proliferation with IC50 values comparable to those required for inhibition of P11274 - P00519 ( 0.3 to 20 nmol/L ) . The activity of ponatinib against the P36888 -ITD mutant , found in up to 30 % of acute myeloid leukemia ( AML ) patients , was particularly notable . In MV4-11 ( P36888 -ITD(+/+) ) but not RS4;11 ( P36888 -ITD(-/-) ) AML cells , ponatinib inhibited P36888 signaling and induced apoptosis at concentrations of less than 10 nmol/L . In an MV4-11 mouse xenograft model , once daily oral dosing of ponatinib led to a dose-dependent inhibition of signaling and tumor regression . DB08901 inhibited viability of primary leukemic blasts from a P36888 -ITD positive AML patient ( IC50 4 nmol/L ) but not those isolated from 3 patients with AML expressing native P36888 . Overall , these results support the investigation of ponatinib in patients with P36888 -ITD-driven AML and other hematologic malignancies driven by P10721 , P11362 , or P09619 α . DB01095 inhibits growth and alters the malignant phenotype of the P13671 glioma cell line . BACKGROUND : DB01095 is a member of the family of P04035 inhibitors ( statins ) extensively used in medical practice . Increasing evidence suggests that fluvastatin may be implicated in suppression of cancer growth and development . The aim of the present study was to investigate the anti-cancer potential of fluvastatin in P13671 rat malignant glioma cells . METHODS : First , the effects of fluvastatin on cell viability ( MTT assay ) , proliferation ( BrdU assay ) , cell morphology , and cytoskeleton were examined . Subsequently , its effect on extracellular signal regulated kinase 1 and 2 ( P27361 /2 ) and P45983 and 2 ( JNK 1/2 ) expression was estimated by Western blot . Finally , the influence of fluvastatin on cell migration and production of P14780 and P15692 was determined using a wound-healing assay and ELISA test , respectively . RESULTS : The results obtained showed that fluvastatin had a remarkable inhibitory and cytotoxic effect on tumor P13671 cells ( IC(50) = 8.6 μM , 48 h ) , but did not inhibit the growth of normal neuronal cells . The concentrations from 1 to 10 μM induced marked morphologic alterations typical for apoptosis including shrinkage of cytoplasm , chromatin condensation , and nucleus breakdown . CONCLUSION : The inhibitory effects of fluvastatin on cell proliferation seemed to be associated with decreased p- P27361 /2 expression , upregulation of p- P45983 /2 , and reduction in the P14780 and P15692 concentrations in culture media . The high anticancer ( antiproliferative , proapoptotic , antiinvasive ) activity of fluvastatin and lack of its toxicity against normal cells indicate a potential use of this statin in the treatment of malignant glioma . Mast cells and eosinophils in mastocytosis , chronic eosinophilic leukemia , and non-clonal disorders . Mast cells and eosinophils often travel in the same biologic circles . In non-clonal states , such as allergic and inflammatory conditions , cell-to-cell contact and the pleiotropic actions of multiple cytokines and chemokines , derived from local tissues or mast cells themselves , foster the co-recruitment of these cells to the same geographic cellular niche . While eosinophils and mast cells serve critical roles as part of the host immune response and in maintenance of normal homeostasis , these cell types can undergo neoplastic transformation due to the development of clonal molecular abnormalities that arise in early hematopoietic progenitors . The dysregulated tyrosine kinases , D816V P10721 and Q6UN15 - P16234 , are the prototypic oncogenic lesions resulting in systemic mastocytosis ( SM ) and chronic eosinophilic leukemia , respectively . We review the pathobiology of these myeloproliferative neoplasms ( MPNs ) with a focus on the relationship between mast cells and eosinophils , and discuss murine models , which further elucidate how the phenotype of these diseases can be influenced by stem cell factor ( P21583 ) and expression of the potent eosinophilopoietic cytokine , interleukin-5 ( P05113 ) . Therapy of SM and Q6UN15 - P16234 -positive disease and the prognostic relevance of increased peripheral blood and tissue mast cells in hematolymphoid malignancies will also be addressed . Synergistic growth-inhibitory effects of ponatinib and midostaurin ( PKC412 ) on neoplastic mast cells carrying P10721 D816V . Patients with advanced systemic mastocytosis , including mast cell leukemia , have a poor prognosis . In these patients , neoplastic mast cells usually harbor the P10721 mutant D816V that confers resistance against tyrosine kinase inhibitors . We examined the effects of the multi-kinase blocker ponatinib on neoplastic mast cells and investigated whether ponatinib acts synergistically with other antineoplastic drugs . DB08901 was found to inhibit the kinase activity of P10721 G560V and P10721 D816V in the human mast cell leukemia cell line HMC-1 . In addition , ponatinib was found to block Lyn- and P42229 activity in neoplastic mast cells . DB08901 induced growth inhibition and apoptosis in HMC-1.1 cells ( P10721 G560V(+) ) and HMC-1.2 cells ( P10721 G560V(+)/ P10721 D816V(+) ) as well as in primary neoplastic mast cells . The effects of ponatinib were dose-dependent , but higher IC50-values were obtained in HMC-1 cells harboring P10721 D816V than in those lacking P10721 D816V . In drug combination experiments , ponatinib was found to synergize with midostaurin in producing growth inhibition and apoptosis in HMC-1 cells and primary neoplastic mast cells . The ponatinib+midostaurin combination induced substantial inhibition of P10721 - , Lyn- , and P42229 activity , but did not suppress Btk . We then applied a Btk short interfering RNA and found that Btk knockdown sensitizes HMC-1 cells against ponatinib . Finally , we were able to show that ponatinib synergizes with the Btk-targeting drug dasatinib to produce growth inhibition in HMC-1 cells . In conclusion , ponatinib exerts major growth-inhibitory effects on neoplastic mast cells in advanced systemic mastocytosis and synergizes with midostaurin and dasatinib in inducing growth arrest in neoplastic mast cells . Where exactly does ponatinib fit in chronic myelogenous leukemia ? DB08901 holds a unique place in the spectrum of drugs in use for the treatment of chronic myelogenous leukemia . It is perhaps the most active tyrosine kinase inhibitor ( TKI ) among those currently licensed ; 51 % of patients resistant to or intolerant of second-generation TKIs experienced a major cytogenetic response and 70 % of patients with the highly resistant T315I P11274 - P00519 mutation experienced a major cytogenetic response . However , 1 year after its accelerated approval by the FDA , and midway through its phase III pivotal trial , a high number of vascular occlusive events began to be reported . The FDA put a partial clinical hold on the drug and the phase III trial was halted . Dose-reduction recommendations were made , and the drug is now used in patients for whom no alternative TKI is available and those who have the T315I mutation . Currently , the substantial and durable responses that this drug provides are difficult to balance against the late-in-course vascular occlusive events . The hope is that ongoing research into the mechanism of presumed endothelial damage will provide a better understanding of how to position this drug for optimal use . Efficacy of ponatinib against P00519 tyrosine kinase inhibitor-resistant leukemia cells . Because a substantial number of patients with chronic myeloid leukemia acquire resistance to P00519 tyrosine kinase inhibitors ( TKIs ) , their management remains a challenge . DB08901 , also known as DB08901 , is an oral multi-targeted TKI . DB08901 is currently being investigated in a pivotal phase 2 clinical trial . In the present study , we analyzed the molecular and functional consequences of ponatinib against imatinib- or nilotinib-resistant ( R ) K562 and Ba/ P13726 cells . The proliferation of imatinib- or nilotinib-resistant K562 cells did not decrease after treatment with imatinib or nilotinib . Src family kinase Lyn was activated . Point mutation Ba/ P13726 cells ( E334V ) were also highly resistant to imatinib and nilotinib . Treatment with ponatinib for 72h inhibited the growth of imatinib- and nilotinib-resistant cells . The phosphorylation of P11274 - P00519 , Lyn , and Crk-L was reduced . This study demonstrates that ponatinib has an anti-leukemia effect by reducing P00519 and Lyn kinase activity and this information may be of therapeutic relevance . The novel P11274 - P00519 and P36888 inhibitor ponatinib is a potent inhibitor of the MDR-associated DB00171 -binding cassette transporter Q9UNQ0 . DB08901 is a novel tyrosine kinase inhibitor with potent activity against P11274 - P00519 with mutations , including T315I , and also against fms-like tyrosine kinase 3 . We tested interactions between ponatinib at pharmacologically relevant concentrations of 50 to 200 nmol/L and the MDR-associated DB00171 -binding cassette ( DB01048 ) proteins P08183 , P33527 , and Q9UNQ0 . DB08901 enhanced uptake of substrates of Q9UNQ0 and P08183 , but not P33527 , in cells overexpressing these proteins , with a greater effect on Q9UNQ0 than on P08183 . DB08901 potently inhibited [ (125)I ] -IAAP binding to Q9UNQ0 and P08183 , indicating binding to their drug substrate sites , with IC(50) values of 0.04 and 0.63 μmol/L , respectively . DB08901 stimulated Q9UNQ0 ATPase activity in a concentration-dependent manner and stimulated P08183 ATPase activity at low concentrations , consistent with it being a substrate of both proteins at pharmacologically relevant concentrations . The ponatinib IC(50) values of P11274 - P00519 -expressing K562 cells transfected with P08183 and Q9UNQ0 were approximately the same as and 2-fold higher than that of K562 , respectively , consistent with ponatinib being a substrate of both proteins , but inhibiting its own transport , and resistance was also attenuated to a small degree by ponatinib-induced downregulation of P08183 and Q9UNQ0 cell-surface expression on resistant K562 cells . DB08901 at pharmacologically relevant concentrations produced synergistic cytotoxicity with P08183 and Q9UNQ0 substrate chemotherapy drugs and enhanced apoptosis induced by these drugs , including daunorubicin , mitoxantrone , topotecan , and flavopiridol , in cells overexpressing these transport proteins . Combinations of ponatinib and chemotherapy drugs warrant further testing . Effects of ketoconazole on the pharmacokinetics of ponatinib in healthy subjects . DB08901 is a P11274 - P00519 tyrosine kinase inhibitor ( TKI ) approved for the treatment of chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia in patients resistant or intolerant to prior TKIs . In vitro studies suggested that metabolism of ponatinib is partially mediated by P08684 . The effects of P08684 inhibition on the pharmacokinetics of ponatinib and its P08684 -mediated metabolite , AP24567 , were evaluated in a single-center , randomized , two-period , two-sequence crossover study in healthy volunteers . Subjects ( N = 22 ) received two single doses ( orally ) of ponatinib 15 mg , once given alone and once coadministered with daily ( 5 days ) ketoconazole 400 mg , a P08684 inhibitor . DB08901 plus ketoconazole increased ponatinib maximum plasma concentration ( C(max) ) and area under the concentration-time curve ( AUC ) compared with ponatinib alone . The estimated mean ratios for AUC0-∞ , AUC0-t , and C(max) indicated increased exposures to ponatinib of 78 % , 70 % , and 47 % , respectively ; exposure to AP24567 decreased by 71 % . Exposure to AP24567 was marginal after ponatinib alone ( no more than 4 % of the exposure to ponatinib ) . These results suggest that caution should be exercised with the concurrent use of ponatinib and strong P08684 inhibitors and that a ponatinib dose decrease to 30 mg daily , from the 45 mg daily starting dose , could be considered . Targeting the P11274 - P00519 signaling pathway in therapy-resistant Philadelphia chromosome-positive leukemia . Beginning with imatinib a decade ago , therapy based on targeted inhibition of the P11274 - P00519 kinase has greatly improved the prognosis for chronic myeloid leukemia ( CML ) patients . The recognition that some patients experience relapse due to resistance-conferring point mutations within P11274 - P00519 sparked the development of the second-generation P00519 kinase inhibitors nilotinib and dasatinib . Collectively , these drugs target most resistant P11274 - P00519 mutants , with the exception of P11274 - P00519 (T315I) . A third wave of advances is now cresting in the form of P00519 kinase inhibitors whose target profile encompasses P11274 - P00519 (T315I) . The leading third-generation clinical candidate for treatment-refractory CML , including patients with the T315I mutation , is ponatinib ( DB08901 ) , a pan- P11274 - P00519 inhibitor that has entered pivotal phase 2 testing . A second inhibitor with activity against the P11274 - P00519 (T315I) mutant , P43146 -2036 , is in phase 1 clinical evaluation . We provide an up-to-date synopsis of P11274 - P00519 signaling pathways , highlight new findings on mechanisms underlying P11274 - P00519 mutation acquisition and disease progression , discuss the use of nilotinib and dasatinib in a first-line capacity , and evaluate ponatinib , P43146 -2036 , and other P00519 kinase inhibitors with activity against P11274 - P00519 (T315I) in the development pipeline . DB08901 is a pan- P11274 - P00519 kinase inhibitor : MD simulations and SIE study . P11274 - P00519 kinase domain inhibition can be used to treat chronic myeloid leukemia . The inhibitors such as imatinib , dasatinib and nilotinib are effective drugs but are resistant to some P11274 - P00519 mutations . The pan- P11274 - P00519 kinase inhibitor ponatinib exhibits potent activity against native , T315I , and all other clinically relevant mutants , and showed better inhibition than the previously known inhibitors . We have studied the molecular dynamics simulations and calculated solvated interaction energies of native and fourteen mutant P11274 - P00519 kinases ( M244V , G250E , Q252H , Y253F , Y253H , E255K , E255V , T315A , T315I , F317L , F317V , M351T , F359V and H396P ) complexed with ponatinib . These studies revealed that the interactions between ponatinib and individual residues in P11274 - P00519 kinase are also affected due to the remote residue mutations . We report that some residues , Met244 , Lys245 , Gln252 , Gly254 , Leu370 and Leu298 do not undergo any conformational changes , while the fluctuations in residues from P-loop , β3- , β5- strands and αC- helix are mainly responsible for ponatinib binding to native and all mutant P11274 - P00519 kinases . Our work provides the molecular mechanisms of native and mutant P11274 - P00519 kinases inhibition by ponatinib at atomic level that has not been studied before . Serotonin transporter interacts with the PDGFβ receptor in DB00102 -induced signaling and mitogenesis in pulmonary artery smooth muscle cells . The serotonin transporter ( P31645 ) and the platelet-derived growth factor receptor ( P09619 ) have been implicated in both clinical and experimental pulmonary hypertension ( PH ) and the facilitation of pulmonary artery smooth muscle cell ( PASMC ) growth . To gain a better understanding of the possible relationship of these two cell surface molecules we have explored interactions between P31645 and P09619 . We have previously demonstrated that P31645 transactivates PDGFRβ in serotonin-stimulated PASMC proliferation . We now provide evidence for a role for P31645 in DB00102 signaling and PASMC proliferation by using pharmacological inhibitors , genetic ablation , and construct overexpression of P31645 . The results show that four tested P31645 blockers dose dependently inhibit PDGF-stimulated human and bovine PASMC proliferation with comparable efficacy to that of P09619 inhibitors , whereas P28222 or 5- Q13049 receptor inhibitors had no effect . Combinations of the P31645 and P09619 inhibitors led to synergistic/additive inhibition . Similarly , PDGF-induced PASMC proliferation was attenuated by small interfering RNA downregulation of P31645 . Inhibition of P31645 in PASMCs attenuated PDGF-induced phosphorylation of PDGFRβ , Akt , and p38 but not Erk . Overexpression of P31645 in HEK293 cells led to enhanced Akt phosphorylation by PDGF , which was blunted by a P31645 PDZ motif mutant , indicating the mechanistic need for the PDZ motif of P31645 in PDGF signaling . Furthermore , coimmunoprecipitation experiments showed that P31645 and PDGFRβ become physically associated upon PDGF stimulation . In total , the data show for the first time an important interactive relationship between P31645 and the PDGFRβ in the production of PASMC proliferation triggered by PDGF that may be important in PH . Combined targeting of P21802 and P42345 by ponatinib and ridaforolimus results in synergistic antitumor activity in P21802 mutant endometrial cancer models . PURPOSE : Activating mutations in P21802 have been identified as potential therapeutic targets in endometrial cancer , typically occurring alongside genetic alterations that disrupt the P42345 pathway , such as P60484 loss . These observations suggest that the P42345 pathway may act in concert with oncogenic P21802 to drive endometrial cancer growth in a subset of patients . The aim of this study was to examine the therapeutic potential of a rational drug combination based on the simultaneous targeting of mutant- P21802 and P42345 -driven signaling pathways in endometrial cancer cells . METHODS : DB08901 is an oral multitargeted kinase inhibitor that potently inhibits all 4 members of the FGFR family . Ridaforolimus is a selective inhibitor of P42345 that has demonstrated positive clinical activity in endometrial cancer . The combinatorial effects of ponatinib and ridaforolimus on growth of endometrial cancer models , and their modes of action , were evaluated in vitro and in vivo . RESULTS : The combination of ponatinib and ridaforolimus had a synergistic effect on the in vitro growth of endometrial lines bearing an activating P21802 mutation , irrespective of P60484 status . Concomitant inhibition of both P21802 and P42345 signaling pathways was observed , with simultaneous blockade resulting in enhanced cell cycle arrest . DB08901 and ridaforolimus each demonstrated inhibition of tumor growth in vivo , but dual inhibition by the combination of agents resulted in superior efficacy and induced tumor regression in an endometrial xenograft . CONCLUSIONS : These encouraging preclinical findings suggest the inhibition of both P21802 and P42345 by the ponatinib-ridaforolimus combination may provide a new therapeutic strategy to treat advanced endometrial cancers with dual pathway dysregulation . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . [ Is DB08901 ( DB08901 ) the next treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia ? ] . Distinct clinicopathologic acute lymphoblastic leukemia ( ALL ) entities have been identified , resulting in the adoption of risk-oriented treatment approaches . In Philadelphia chromosome-positive ( Ph(+) ) ALL , the optimal treatment requires the addition of P11274 - P00519 tyrosine kinase inhibitors , as imatinib . However , the outcome remains poor in absence of allogeneic stem cell transplantation , and novel agents are desperately required . Resistance attributable to kinase domain mutations can lead to relapse despite the development of second-generation compounds , including dasatinib and nilotinib . Despite these therapeutic options , the cross-resistant P11274 - P00519 ( T315I ) mutation remains a major clinical challenge . The first evaluations of DB08901 present this drug as a potent multi-targeted kinase inhibitor active against T315I and all other P11274 - P00519 mutants . DB08901 could be the next treatment of choice in hematological malignancies with Philadelphia-positive chromosome , particularly Ph(+) ALL known for its frequent occurrence of T315I mutation . [ Chronic myeloid leukemia ] . More than 10 years have passed since imatinib as a first developed P11274 - P00519 tyrosine kinase inhibitor ( TKI ) introduced in treatment of patients with chronic myeloid leukemia ( CML ) . In globally , there are tremendous numbers of patients on imatinib therapy . Based upon randomized trials comparing second generation TKIs such as dasatinib and nilotinib versus imatinib , both TKIs produce faster and deeper response than imatinib and they can be selected as first-line therapy for newly diagnosed chronic phase of CML ( CP-CML ) as imatinib . Bosutinib is a potent for imatinib resistant/intolerant CP-CML and can be used as second or third-line therapy . DB08901 is the only clinically available TKI that has activity against the T315 mutation that is resistant to all other TKIs . Currently , a choice among these potent TKIs should take into consideration the drug side effect profiles and the patient 's comorbidities . Multiple checkpoint breach of B cell tolerance in Rasgrp1-deficient mice . Lymphopenic hosts offer propitious microenvironments for expansion of autoreactive B and T cells . Despite this , many lymphopenic hosts do not develop autoimmune disease , suggesting that additional factors are required for breaching self-tolerance in the setting of lymphopenia . Mice deficient in guanine nucleotide exchange factor Rasgrp1 develop a lymphoproliferative disorder with features of human systemic lupus erythematosus . Early in life , Rasgrp1-deficient mice have normal B cell numbers but are T lymphopenic , leading to defective homeostatic expansion of P01730 T cells . To investigate whether B cell-intrinsic mechanisms also contribute to autoimmunity , Rasgrp1-deficient mice were bred to mice containing a knockin autoreactive P11274 transgene ( 564Igi ) , thereby allowing the fate of autoreactive B cells to be assessed . During B cell development , the frequency of receptor-edited 564Igi B cells was reduced in Rasrp1-deficient mice compared with Rasgrp1-sufficient littermate control mice , suggesting that tolerance was impaired . In addition , the number of 564Igi transitional B cells was increased in Rasgrp1-deficient mice compared with control mice . Immature 564Igi B cells in bone marrow and spleen lacking RasGRP1 expressed lower levels of Bim mRNA and protein , suggesting that autoreactive B cells elude clonal deletion during development . Concomitant with increased serum autoantibodies , Rasgrp1-deficient mice developed spontaneous germinal centers at 8-10 wk of age . The frequency and number of 564Igi B cells within these germinal centers were significantly increased in Rasgrp1-deficient mice relative to control mice . Taken together , these studies suggest that autoreactive B cells lacking Rasgrp1 break central and peripheral tolerance through both T cell-independent and -dependent mechanisms . Resistant mutations in CML and Ph(+)ALL - role of ponatinib . In 2012 , ponatinib ( Iclusig(®) ) , an orally available pan- P11274 - P00519 tyrosine kinase inhibitor ( TKI ) developed by ARIAD Pharmaceuticals , Inc. , was approved by the US Food and Drug Administration for use in resistant or intolerant chronic myeloid leukemia ( CML ) and Philadelphia chromosome-positive acute lymphoblastic leukemia ( Ph(+)ALL ) . DB08901 is the only approved TKI capable of inhibiting P11274 - P00519 with the gatekeeper T315I kinase domain mutation , known to be the cause for 20 % of resistant or relapsed CML cases . In 2013 , ponatinib sales were temporarily suspended due to serious side effects seen in nearly 12 % of the patient population . These side effects are thought to stem from the potent nature and pan-activity of this TKI . ARIAD Pharmaceuticals , Inc. has since been permitted to resume sales and marketing of ponatinib to a limited patient population with an expanded black box warning . In the following review , the use of ponatinib in CML and Ph(+)ALL will be discussed . Mechanisms of resistance in CML are discussed , which provide insight and background into the need for this third generation TKI , followed by the molecular design and pharmacology of ponatinib , which lead to its success as a therapeutic . Finally , the efficacy , safety , and tolerability of ponatinib will be highlighted , including summaries of the important clinical trials involving ponatinib as well as its current place in therapy . DB08901 for the treatment of chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia . DB08901 is a novel , next-generation , small-molecule tyrosine kinase inhibitor with potent activity against the P11274 - P00519 fusion oncogene as well as all other P00519 kinase domain mutations that confer resistance to earlier generation tyrosine kinase inhibitors . Due to its unique structure , it is the only tyrosine kinase inhibitor with the capability to counter the highly resistant T315I or gate-keeper mutation in leukemic cells that express the Philadelphia chromosome . This review will focus on the preclinical pharmacology , pharmacokinetics and clinical utility of ponatinib in the treatment of chronic myeloid leukemia and Philadelphia chromosome-positive adult acute lymphoblastic leukemia . All tyrosine kinase inhibitor-resistant chronic myelogenous cells are highly sensitive to ponatinib . The advent of tyrosine kinase inhibitor ( TKI ) therapy has considerably improved the survival of patients suffering chronic myelogenous leukemia ( CML ) . Indeed , inhibition of P11274 - P00519 by imatinib , dasatinib or nilotinib triggers durable responses in most patients suffering from this disease . Moreover , resistance to imatinib due to kinase domain mutations can be generally circumvented using dasatinib or nilotinib , but the multi-resistant T315I mutation that is insensitive to these TKIs , remains to date a major clinical problem . In this line , ponatinib ( DB08901 ) has emerged as a promising therapeutic option in patients with all kinds of P11274 - P00519 mutations , especially the T315I one . However and surprisingly , the effect of ponatinib has not been extensively studied on imatinib-resistant CML cell lines . Therefore , in the present study , we used several CML cell lines with different mechanisms of resistance to TKI to evaluate the effect of ponatinib on cell viability , apoptosis and signaling . Our results show that ponatinib is highly effective on both sensitive and resistant CML cell lines , whatever the mode of resistance and also on BaF3 murine B cells carrying native P11274 - P00519 or T315I mutation . We conclude that ponatinib could be effectively used for all types of TKI-resistant patients . DB08901 inhibits polyclonal drug-resistant P10721 oncoproteins and shows therapeutic potential in heavily pretreated gastrointestinal stromal tumor ( GIST ) patients . PURPOSE : P10721 is the major oncogenic driver of gastrointestinal stromal tumors ( GIST ) . Imatinib , sunitinib , and regorafenib are approved therapies ; however , efficacy is often limited by the acquisition of polyclonal secondary resistance mutations in P10721 , with those located in the activation ( A ) loop ( exons 17/18 ) being particularly problematic . Here , we explore the P10721 -inhibitory activity of ponatinib in preclinical models and describe initial characterization of its activity in patients with GIST . EXPERIMENTAL DESIGN : The cellular and in vivo activities of ponatinib , imatinib , sunitinib , and regorafenib against mutant P10721 were evaluated using an accelerated mutagenesis assay and a panel of engineered and GIST-derived cell lines . The ponatinib- P10721 costructure was also determined . The clinical activity of ponatinib was examined in three patients with GIST previously treated with all three FDA-approved agents . RESULTS : In engineered and GIST-derived cell lines , ponatinib potently inhibited P10721 exon 11 primary mutants and a range of secondary mutants , including those within the A-loop . DB08901 also induced regression in engineered and GIST-derived tumor models containing these secondary mutations . In a mutagenesis screen , 40 nmol/L ponatinib was sufficient to suppress outgrowth of all secondary mutants except V654A , which was suppressed at 80 nmol/L . This inhibitory profile could be rationalized on the basis of structural analyses . DB08901 ( 30 mg daily ) displayed encouraging clinical activity in two of three patients with GIST . CONCLUSION : DB08901 possesses potent activity against most major clinically relevant P10721 mutants and has demonstrated preliminary evidence of activity in patients with refractory GIST . These data strongly support further evaluation of ponatinib in patients with GIST . Peripheral blood gene expression of P33681 and P10747 family members associated with tumor progression and microscopic lymphovascular invasion in colon cancer patients . PURPOSE : To associate the global gene expression of P33681 / P10747 family transcripts with pathologic features of colon cancer , we determined the P33681 / P10747 family transcripts in peripheral blood mononuclear cells ( PBMCs ) from normal subjects and patients with adenomatous polyps and colon cancer , and correlated the results with pathologic features of colon cancer . METHODS : PBMCs from age-matched normal subjects and patients with adenomatous polyps and colon cancer were analyzed for peripheral blood transcripts ( PBTs ) of P33681 / P10747 family using real-time PCR . Differences in expression levels of P33681 / P10747 PBTs across all cancer stages and between colon cancer patients with or without microscopic lymphovascular invasion ( LVI ) were analyzed . RESULTS : The results showed a significant upregulation of PBTs of co-inhibitory molecules such as Q5ZPR3 and P18621 and a significant P10721 downregulation of co-stimulatory molecules including P10747 and Q9Y6W8 in colon cancer patients . Furthermore , the increase of Q5ZPR3 P10721 was strongly associated with tumor invasion ( P = 0.025 ) and advanced TNM stages ( P = 0.019 ) , whereas the decline of co-stimulatory ligand O75144 P10721 was related to regional lymph node metastasis ( P = 0.028 ) and aggressive tumor invasion ( P = 0.031 ) . In addition , the ratios of P10721 expression of P10747 family to P33681 family such as P16410 to O75144 and P18621 to O75144 were significantly higher in colon cancer patients with microscopic LVI than in those without LVI ( P = 0.001 and P = 0.016 , respectively ) . CONCLUSIONS : Our results suggest that P33681 / P10747 family PBTs may serve as valuable markers reflecting the pathological features of colon cancer . DB08901 as targeted therapy for P11362 fusions associated with the 8p11 myeloproliferative syndrome . The 8p11 myeloproliferative syndrome is a rare , aggressive myeloproliferative neoplasm characterized by constitutively active P11362 fusion proteins that arise from specific chromosomal translocations and which drive aberrant proliferation . Although P11362 inhibitors have shown in vitro activity against P11362 fusions , none are in use clinically and there is a need to assess additional compounds as potential therapy . Here we use cell lines and primary cells to investigate ponatinib ( DB08901 ) . DB08901 -treated Ba/ P13726 cells transformed by Q9UBW7 - P11362 and P11274 - P11362 and the Q9NVK5 - P11362 positive KG1A cell line showed reduced proliferation and decreased survival when compared to control cells . Inhibition induced apoptosis and reduced phosphorylation of the P11362 fusion proteins and substrates . DB08901 -treated cells from 8p11 myeloproliferative syndrome patients ( n=5 ) showed reduced colony growth compared to controls . In one evaluable patient , ponatinib specifically reduced numbers of P11362 -fusion gene positive colonies . DB08901 , therefore , shows considerable promise for the treatment of patients with 8p11 myeloproliferative syndrome . Discovery of 3-[2-(imidazo[1,2-b]pyridazin-3-yl)ethynyl]-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide ( DB08901 ) , a potent , orally active pan-inhibitor of breakpoint cluster region-abelson ( P11274 - P00519 ) kinase including the T315I gatekeeper mutant . In the treatment of chronic myeloid leukemia ( CML ) with P11274 - P00519 kinase inhibitors , the T315I gatekeeper mutant has emerged as resistant to all currently approved agents . This report describes the structure-guided design of a novel series of potent pan-inhibitors of P11274 - P00519 , including the T315I mutation . A key structural feature is the carbon-carbon triple bond linker which skirts the increased bulk of Ile315 side chain . Extensive SAR studies led to the discovery of development candidate 20g ( DB08901 ) , which inhibited the kinase activity of both native P11274 - P00519 and the T315I mutant with low nM IC(50)s , and potently inhibited proliferation of corresponding Ba/ P13726 -derived cell lines . Daily oral administration of 20g significantly prolonged survival of mice injected intravenously with P11274 - P00519 (T315I) expressing Ba/ P13726 cells . These data , coupled with a favorable ADME profile , support the potential of 20g to be an effective treatment for CML , including patients refractory to all currently approved therapies . Multifaceted link between cancer and inflammation . Increasing evidence from epidemiological , preclinical and clinical studies suggests that dysregulated inflammatory response plays a pivotal role in a multitude of chronic ailments including cancer . The molecular mechanism(s) by which chronic inflammation drives cancer initiation and promotion include increased production of pro-inflammatory mediators , such as cytokines , chemokines , reactive oxygen intermediates , increased expression of oncogenes , P35354 ( cyclo-oxygenase-2 ) , 5- P28300 ( P09917 ) and MMPs ( matrix metalloproteinases ) , and pro-inflammatory transcription factors such as NF-κB ( nuclear factor κB ) , P40763 ( signal transducer and activator of transcription 3 ) , AP-1 ( activator protein 1 ) and HIF-1α ( hypoxia-inducible factor 1α ) that mediate tumour cell proliferation , transformation , metastasis , survival , invasion , angiogenesis , chemoresistance and radioresistance . These inflammation-associated molecules are activated by a number of environmental and lifestyle-related factors including infectious agents , tobacco , stress , diet , obesity and alcohol , which together are thought to drive as much as 90 % of all cancers . The present review will focus primarily on the role of various inflammatory intermediates responsible for tumour initiation and progression , and discuss in detail the critical link between inflammation and cancer . P62158 protects cells from death under normal growth conditions and mitogenic starvation but plays a mediating role in cell death upon B-cell receptor stimulation . P62158 ( P62158 ) is the main intracellular Ca2+ sensor protein responsible for mediating Ca2+ triggered processes . Chicken DT40 lymphoma B cells express P62158 from the two genes , CaMI and CaMII . Here we report the phenotypes of DT40 cells with the CaMII gene knocked out . The disruption of the CaMII gene causes the intracellular P62158 level to decrease by 60 % . CaMII-/- cells grow more slowly and die more frequently as compared to wild type ( wt ) cells but do not exhibit significant differences in their cell cycle profile . Both phenotypes are more pronounced at reduced serum concentrations . Upon stimulation of the B-cell receptor ( P11274 ) , the resting Ca2+ levels remain elevated after the initial transient in CaMII-/- cells . Despite higher Ca2+ resting levels , the CaMII-/- cells are partially protected from P11274 induced apoptosis indicating that P62158 plays a dual role in apoptotic processes . H2O2 production downstream of P36888 is mediated by P13498 in the endoplasmic reticulum and is required for P42229 signalling . The internal tandem duplication ( ITD ) of the juxtamembrane region of the P36888 receptor has been associated with increased reactive oxygen species ( ROS ) generation in acute myeloid leukemia ( AML ) . How this elevated level of ROS contributes to the leukemic phenotype , however , remains poorly understood . In this work we show that ROS in the P36888 -ITD expressing AML cell line MV4-11 is reduced by treatment with PKC412 , an inhibitor of P36888 , DPI , a flavoprotein inhibitor , and VAS2870 , a Nox specific inhibitor , suggesting that ROS production is both P36888 and NADPH oxidase dependent . The majority of these ROS co-localize to the endoplasmic reticulum ( ER ) , as determined with the H(2)O(2)-specific aryl-boronate dye Peroxyorange 1 , which also corresponds to co-localization of P13498 . Moreover , knocking down P13498 dramatically reduces H(2)O(2) after 24 hours in the ER , without affecting mitochondrial ROS . Significantly , the P36888 inhibitor PKC412 reduces H(2)O(2) in P36888 -ITD expressing cell lines ( MV4-11 , MOLM-13 ) through reduction of P13498 over 24 hours . Reduced P13498 is achieved by proteasomal degradation and is prevented upon GSK3-β inhibition . Knockdown of P13498 resulted in reduced P42229 signalling and reduced Pim-1 levels in the cells after 24 hours . Thus , we have shown that P36888 driven H(2)O(2) production in AML cells is mediated by P13498 and is critical for P42229 signalling . DB08901 : a third-generation inhibitor for the treatment of CML . The establishment of imatinib as the standard therapy for CML marked the beginning of a new era of treatment . Due to occurring intolerance and resistance against the drug , developing newer inhibitors was promoted . This led to the second-generation inhibitors dasatinib , nilotinib and bosutinib . Despite all achieved improvement , all first- and second-generation inhibitors are ineffective against the P11274 - P00519 T315I " gatekeeper " mutation . In order to overcome this issue and to further improve the inhibitory effect , the third-generation inhibitor ponatinib was developed . Various clinical trials have been launched to study the effect of ponatinib in the clinical setting . Based on positive phase 1 and phase 2 trials , ponatinib was approved for the second-line treatment of CML and Ph+ ALL in December 2012 in the United States and in July 2013 in the European Union . Further trials investigate the potential effect of ponatinib in kinase-dependent subgroups of other malignancies . In conclusion , ponatinib has proved to be a powerful P11274 - P00519 inhibitor , which exhibits clinical activity both in P11274 - P00519 wild-type and mutant CML , including activity against the T315I mutation . Despite previous TKI failure , chronic-phase CML patients can achieve sustained remissions using the novel drug , offering a new therapeutic option in the treatment for CML . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . DB08901 induces apoptosis in imatinib-resistant human mast cells by dephosphorylating mutant D816V P10721 and silencing β-catenin signaling . Gain-of-function mutations of membrane receptor tyrosine kinase P10721 , especially gatekeeper D816V point mutation in P10721 , render kinase autoactivation , disease progression , and poor prognosis . D816V P10721 is found in approximately 80 % of the patients with systemic mastocytosis , and is resistant to the first and second generations of tyrosine kinase inhibitors ( TKI ) . The purpose of this investigation was aimed at exploring whether ponatinib ( DB08901 ) , a novel effective TKI against T315I Bcr-Abl , was active against D816V P10721 . We discovered that ponatinib abrogated the phosphorylation of P10721 harboring either V560G ( sensitive to imatinib ) or D816V mutation ( resistant to imatinib ) and the downstream signaling transduction . DB08901 inhibited the growth of D816V P10721 -expressing cells in culture and nude mouse xenografted tumor . DB08901 triggered apoptosis by inducing the release of cytochrome c and O95831 , downregulation of Mcl-1 . Furthermore , ponatinib abrogated the phosphorylation of β-catenin at the site Y654 , suppressed the translocation of β-catenin , and inhibited the transcription and DNA binding of TCF and the expression of its targets ( e.g. , Q9Y2T1 , c-MYC , and P24385 ) . Moreover , ponatinib was highly active against xenografted D816V P10721 tumors in nude mice and significantly prolonged the survival of mice with aggressive systemic mastocytosis or mast cell leukemia by impeding the expansion and infiltration of mast cells with imatinib-resistant D814Y P10721 . Our findings warrant a clinical trial of ponatinib in patients with systemic mastocytosis harboring D816V P10721 . Discovery of 5-(arenethynyl) hetero-monocyclic derivatives as potent inhibitors of P11274 - P00519 including the T315I gatekeeper mutant . DB08901 ( DB08901 ) was previously identified as a pan- P11274 - P00519 inhibitor that potently inhibits the T315I gatekeeper mutant , and has advanced into clinical development for the treatment of refractory or resistant CML . In this study , we explored a novel series of five and six membered monocycles as alternate hinge-binding templates to replace the 6,5-fused imidazopyridazine core of ponatinib . Like ponatinib , these monocycles are tethered to pendant toluanilides via an ethynyl linker . Several compounds in this series displayed excellent in vitro potency against both native P11274 - P00519 and the T315I mutant . Notably , a subset of inhibitors exhibited desirable PK and were orally active in a mouse model of T315I-driven CML . Novel FGFR inhibitor ponatinib suppresses the growth of non-small cell lung cancer cells overexpressing P11362 . Lung cancer is still the leading cause of cancer-related deaths worldwide . Identifying new oncogenic drivers and developing efficient inhibitors through molecular targeting approaches are crucial for improving therapies . The aim of this study was to investigate whether targeting fibroblast growth factor receptor 1 ( P11362 ) with ponatinib inhibits the cell growth in both established and primary lung cancer cells overexpressing P11362 . Eighty-eight non-small cell lung cancer ( NSCLC ) and paired normal tissue specimens were analyzed by real-time RT-PCR for P11362 gene expression . We identified four cell lines and two newly established primary lung cancer cultures that showed high P11362 expression levels , and evaluated the effect of the novel P11362 inhibitor ponatinib on cell growth . Approximately 50 % ( 30 out of 59 ) NSCLC specimens expressed P11362 > 2-fold compared with their adjacent normal counterparts using quantitative RT-PCR . DB08901 treatment of established NSCLC cell lines expressing higher levels of P11362 resulted in marked cell growth inhibition and suppression of clonogenicity . This growth inhibition was associated with inactivation of P11362 and its downstream targets . P11362 knockdown by shRNA achieved similar results when compared to treatment with ponatinib . Furthermore , ponatinib was able to significantly inhibit the growth of primary lung cancer cultures in vitro . Our data indicate that pharmacological inhibition of P11362 kinase activity with ponatinib may be effective for the treatment of lung cancer patients whose tumors overexpress P11362 . Q9Y6W8 -induced B7h shedding on B cells is inhibited by Q9NYK1 /8 and Q9NR96 . We report in this study that B7h , the ligand for the Q9Y6W8 costimulatory receptor , is rapidly shed from mouse B cells following either Q9Y6W8 binding or P11274 engagement . Shedding occurs through proteolytic cleavage that releases the extracellular Q9Y6W8 -binding region of B7h . Prior exposure of B7h-expressing APCs to Q9Y6W8 -expressing cells inhibits their subsequent ability to costimulate P01579 and P05112 production from P01730 + T cells . Shedding is regulated as Q9NYK1 /8 and Q9NR96 ligands inhibit B7h shedding . A shedding-resistant B7h mutant elicits greater costimulation of P01579 production from P01730 + T cells than does wild-type B7h . These data define shedding of B7h as a novel mechanism for controlling costimulatory signaling by P33681 - P10747 family members that is regulated on B cells by TLR signaling . P50281 as a downstream target of P11274 - P00519 / P00519 interactor 1 signaling : polarized distribution and involvement in P11274 - P00519 -stimulated leukemic cell migration . Membrane-Type 1 Matrix Metalloproteinase Downregulates Fibroblast Growth Factor-2 Binding to the Cell Surface and Intracellular Signaling . Membrane-type 1 matrix metalloproteinase ( P50281 , P50281 ) , a transmembrane proteinase with an extracellular catalytic domain and a short cytoplasmic tail , degrades extracellular matrix components and controls diverse cell functions through proteolytic and non-proteolytic interactions with extracellular , intracellular , and transmembrane proteins . Here we show that in tumor cells P50281 downregulates fibroblast growth factor-2 ( P09038 ) signaling by reducing the amount of P09038 bound to the cell surface with high and low affinity . P09038 induces weaker activation of P27361 /2 Q96HU1 kinase in P50281 expressing cells than in cells devoid of P50281 . This effect is abolished in cells that express proteolytically inactive P50281 but persists in cells expressing P50281 mutants devoid of hemopexin-like or cytoplasmic domain , showing that P09038 signaling is downregulated by P50281 proteolytic activity . P50281 expression results in downregulation of P11362 and -4 , and in decreased amount of cell surface-associated P09038 . In addition , P50281 strongly reduces the amount of P09038 bound to the cell surface with low affinity . Because P09038 association with low-affinity binding sites is a prerequisite for binding to its high-affinity receptors , downregulation of low-affinity binding to the cell surface results in decreased P09038 signaling . Consistent with this conclusion , P09038 induction of tumor cell migration and invasion in vitro is stronger in cells devoid of MT1- MMP than in P50281 expressing cells . Thus , P50281 controls P09038 signaling by a proteolytic mechanism that decreases the cell 's biological response to P09038 . DB08901 suppresses the development of myeloid and lymphoid malignancies associated with P11362 abnormalities . Myeloid and lymphoid malignancies associated with fibroblast growth factor receptor-1 ( P11362 ) abnormalities are characterized by constitutively activated P11362 kinase and rapid transformation to acute myeloid leukemia and lymphoblastic lymphoma . Molecular targeted therapies have not been widely used for stem cell leukemia/lymphoma ( SCLL ) . DB08901 ( DB08901 ) , which potently inhibits native and mutant P11274 - P00519 , also targets the FGFR family . Using murine BaF3 cells , stably transformed with six different P11362 fusion genes , as well as human KG1 cells expressing activated chimeric P11362 and five newly established murine SCLL cell lines , we show that ponatinib ( < 50 nM ) can effectively inhibit phosphoactivation of the fusion kinases and their downstream effectors , such as PLCγ , Stat5 and Src . DB08901 also significantly extended survival of mice transplanted with different SCLL cell lines . DB08901 administered at 30 mg/kg daily also significantly delayed , or even prevented , tumorigenesis of KG1 cells in xenotransplanted mice . Furthermore , we demonstrate that ponatinib specifically inhibits cell growth and clonogenicity of normal human P28906 + progenitor cells transformed by chimeric P11362 fusion kinases . Overall , our data provide convincing evidence to suggest that pharmacologic inhibition of P11362 fusion kinases with ponatinib is likely to be beneficial for patients with SCLL and perhaps for other human disorders associated with dysregulated P11362 activity . Up-regulation of the phosphoinositide 3-kinase pathway in human lamina propria T lymphocytes . Human intestinal lamina propria T lymphocytes ( LPT ) , when investigated ex vivo , exhibit functional properties profoundly different from those of peripheral blood T lymphocytes ( P10721 ) . One prominent feature represents their enhanced sensitivity to P06729 stimulation when compared to P10721 . Given that LPT are hyporesponsive to T cell receptor ( TCR ) /CD3 stimulation , an alternative activation mode , as mimicked by P06729 triggering in vitro , may be functional in mucosal inflammation in vivo . This study provides insight into signalling events associated with the high P06729 responsiveness of LPT . When compared to P10721 , LPT show an increased activation of the phosphoinositide 3/protein kinase B/glycogen synthase kinase 3beta ( P19957 -kinase/AKT/GSK-3beta ) pathway in response to P06729 stimulation . Evidence is provided that up-regulation of this pathway contributes to the enhanced P06729 -induced cytokine production in LPT . Given the importance of TCR-independent stimulation for the initiation of intestinal immune responses analysis of signalling pathways induced by ' co-stimulatory ' receptors may provide valuable information for therapeutic drug design . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Structural mechanisms determining inhibition of the collagen receptor Q08345 by selective and multi-targeted type II kinase inhibitors . The discoidin domain receptors ( DDRs ) , Q08345 and Q16832 , form a unique subfamily of receptor tyrosine kinases that are activated by the binding of triple-helical collagen . Excessive signaling by Q08345 and Q16832 has been linked to the progression of various human diseases , including fibrosis , atherosclerosis and cancer . We report the inhibition of these unusual receptor tyrosine kinases by the multi-targeted cancer drugs imatinib and ponatinib , as well as the selective type II inhibitor Q08345 -IN-1 . DB08901 is identified as the more potent molecule , which inhibits Q08345 and Q16832 with an IC50 of 9nM . Co-crystal structures of human Q08345 reveal a DFG-out conformation ( DFG , DB00128 - DB00120 - DB00145 ) of the kinase domain that is stabilized by an unusual salt bridge between the activation loop and αD helix . Differences to Abelson kinase ( P00519 ) are observed in the Q08345 P-loop , where a β-hairpin replaces the cage-like structure of P00519 . P-loop residues in Q08345 that confer drug resistance in P00519 are therefore accommodated outside the DB00171 pocket . Whereas imatinib and ponatinib bind potently to both the DDR and P00519 kinases , the hydrophobic interactions of the P00519 P-loop appear poorly satisfied by Q08345 -IN-1 suggesting a structural basis for its Q08345 selectivity . Such inhibitors may have applications in clinical indications of Q08345 and Q16832 overexpression or mutation , including lung cancer . c- P05412 promotes P11274 - P00519 -induced lymphoid leukemia by inhibiting methylation of the 5' region of Cdk6 . The transcription factor c- P05412 and its upstream kinase P45983 have been implicated in P11274 - P00519 -induced leukemogenesis . P45983 has been shown to regulate P10415 expression , thereby altering leukemogenesis , but the impact of c- P05412 remained unclear . In this study , we show that P45983 and c- P05412 promote leukemogenesis via separate pathways , because lack of c- P05412 impairs proliferation of p185( P11274 - P00519 )-transformed cells without affecting their viability . The decreased proliferation of c-Jun(Δ/Δ) cells is associated with the loss of cyclin-dependent kinase 6 ( Q00534 ) expression . In c-Jun(Δ/Δ) cells , Q00534 expression becomes down-regulated upon P11274 - P00519 -induced transformation , which correlates with CpG island methylation within the 5' region of Cdk6 . We verified the impact of Cdk6 deficiency using Cdk6(-/-) mice that developed P11274 - P00519 -induced B-lymphoid leukemia with significantly increased latency and an attenuated disease phenotype . In addition , we show that reexpression of Q00534 in P11274 - P00519 -transformed c-Jun(Δ/Δ) cells reconstitutes proliferation and tumor formation in Nu/Nu mice . In summary , our study reveals a novel function for the activating protein 1 ( AP-1 ) transcription factor c- P05412 in leukemogenesis by antagonizing promoter methylation . Moreover , we identify Q00534 as relevant and critical target of AP-1-regulated DNA methylation on P11274 - P00519 -induced transformation , thereby accelerating leukemogenesis . A new naphthalene glycoside from the sponge-derived fungus Arthrinium sp. ZSDS1- P13726 . One new naphthalene derivative , 1,8-dihydroxynaphthol-1-O-α-l-rhamnopyranoside ( 1 ) , together with one known α-dibenzopyrone , alternariol ( 2 ) , and five xanthones ( 3-7 ) were isolated from the sponge-derived fungus Arthrinium sp . ZSDS1- P13726 . All the isolated compounds ( 1-7 ) were established by comprehensive analysis of the spectral data , especially 1D and 2D NMR ( HMQC and HMBC ) spectra . In the primary bioassays , compound 3 exhibited moderate P35354 inhibition , with IC50 values of 12.2 μM . Targeting wild-type and mutationally activated P22455 in rhabdomyosarcoma with the inhibitor ponatinib ( DB08901 ) . Rhabdomyosarcoma ( RMS ) is the most common childhood soft tissue sarcoma . Despite advances in modern therapy , patients with relapsed or metastatic disease have a very poor clinical prognosis . Fibroblast Growth Factor Receptor 4 ( P22455 ) is a cell surface tyrosine kinase receptor that is involved in normal myogenesis and muscle regeneration , but not commonly expressed in differentiated muscle tissues . Amplification and mutational activation of P22455 has been reported in RMS and promotes tumor progression . Therefore , P22455 is a tractable therapeutic target for patients with RMS . In this study , we used a chimeric Ba/ P13726 P41212 - P22455 construct to test five tyrosine kinase inhibitors reported to specifically inhibit FGFRs in the nanomolar range . We found ponatinib ( DB08901 ) to be the most potent P22455 inhibitor with an IC50 in the nanomolar range . DB08901 inhibited the growth of RMS cells expressing wild-type or mutated P22455 through increased apoptosis . Phosphorylation of wild-type and mutated P22455 as well as its downstream target P40763 was also suppressed by ponatinib . Finally , ponatinib treatment inhibited tumor growth in a RMS mouse model expressing mutated P22455 . Therefore , our data suggests that ponatinib is a potentially effective therapeutic agent for RMS tumors that are driven by a dysregulated P22455 signaling pathway . DB08901 enhances anticancer drug sensitivity in Q5T3U5 -overexpressing cells . The presence of acquired multidrug resistance ( MDR ) is one of the primary impediments to the success of chemotherapy . MDR is often a result of overexpression of DB00171 -binding cassette ( DB01048 ) transporters , which are involved in the extrusion of therapeutic drugs . Recently , it was shown that several ABC transporters could be modulated by specific tyrosine-kinase inhibitors ( TKIs ) . DB08901 , a multi-targeted TKI , inhibits the activity of P11274 - P00519 with very high potency and broad specificity , including the T315I mutation which confers resistance to other TKIs . It was reported that ponatinib was capable of reversing breast cancer resistance protein ( Q9UNQ0 ) - and P-glycoprotein ( P-gp ) -mediated MDR . In the present study , we report for the first time that ponatinib also potentiates the cytotoxicity of widely used therapeutic substrates of Q5T3U5 , such as paclitaxel , docetaxel , vincristine and vinblastine . DB08901 significantly enhances the accumulation of [ 3H ] -paclitaxel in cells expressing Q5T3U5 . Furthermore , accumulation of [ 3H ] -paclitaxel was achieved by inhibition of Q5T3U5 -mediated transport . Ponatinb limited drug export via Q5T3U5 by multiple mechanisms . In addition to inhibition of pump function , ponatinib also downregulated Q5T3U5 protein expression in a time- and concentration-dependent manner . Thus , ponatinib may represent a potential reversal agent for the treatment of MDR and may be useful for combination therapy in MDR cancer patients in clinical practice . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . Potential of ponatinib to treat chronic myeloid leukemia and acute lymphoblastic leukemia . Development of P11274 - P00519 tyrosine kinase inhibitors ( TKIs ) have improved outcomes for patients diagnosed with chronic myeloid leukemia and Philadelphia chromosome positive acute lymphoblastic leukemia . However , resistance or intolerance to these TKIs still leaves some patients without many treatment options . One point mutation in particular , the T315I mutation , has been shown to be resistant to first and second generation TKIs . The third generation TKI , ponatinib , may provide an option for these patients . DB08901 ( Iclusig® ) , an orally available , pan-tyrosine kinase inhibitor has a unique binding mechanism allowing inhibition of P11274 - P00519 kinases , including those with the T315I point mutation . A Phase II study evaluated ponatinib in patients who were resistant or intolerant to nilotinib or dasatinib or patients who had the T315I mutation . In the Phase II study , ponatinib produced a major cytogenetic response in 54 % of chronic phase chronic myeloid leukemia patients . It further achieved major hematologic response in 52 % of patients in the accelerated phase , 31 % of patients in the blast phase , and 41 % of Philadelphia chromosome positive acute lymphoblastic leukemia patients . DB08901 also showed efficacy in patients with the T315I mutation . Serious adverse events included arterial thrombosis , hepatotoxicity , cardiovascular risks , pancreatitis , hemorrhage , fluid retention , myelosuppression , rash , abdominal pain , and embryo-fetal toxicity . Due to the risk of these adverse events and potential drug interactions , the use of ponatinib must be carefully weighed against the benefits in treating patients who have limited treatment options . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . DB00820 , a further innovation in the treatment of sexual dysfunction . In recognition of the large number of sufferers of sexual dysfunction worldwide , and the variety of etiologies of the condition , investigation into effective pharmacological agents has been expanded . One method of intervention is inhibition of the phosphodiesterase type 5 ( O76074 ) enzyme , which has already been exploited with a considerable degree -- though not complete -- success . A number of new agents that inhibit O76074 are under development . Notable among these is tadalafil , which has demonstrated a high level of selectivity for O76074 over the other phosphodiesterases and has shown efficacy in improving erectile function and sexual satisfaction in phase III trials . Throughout the clinical development program for tadalafil , the drug has been well tolerated and without serious side effects . The manufacturer , Lilly Q9Y6W8 , received an approvable letter from the US Food and Drug Administration for use of the drug as a treatment for erectile dysfunction on April 30 , 2002 . Lilly Q9Y6W8 hopes to market tadalafil , with the trade name DB00820 , in the USA in 2003 . Synthesis and biological evaluation of novel pyrrolidine-2,5-dione derivatives as potential antidepressant agents . Part 1 . A series of 3-(1H-indol-3-yl)pyrrolidine-2,5-dione derivatives was synthesized and their biological activity was evaluated . The chemical structures of the newly prepared compounds were confirmed by (1)H NMR , (13)C NMR and P19957 -HRMS spectra data . All tested compounds proved to be potent P08908 receptor and serotonin transporter protein ( P31645 ) ligands . Among them , compounds 15 , 18 , 19 and 30 showed significant affinity for P08908 and P31645 . Computer docking simulations carried out for compounds 15 , 31 and 32 to models of P08908 receptor and P31645 confirm the results of biological tests . Due to high affinity for the P08908 receptor and moderate affinity for P31645 , compounds 31 , 32 , 35 , and 37 were evaluated for their affinity for D2L , P50406 , P34969 and 5- Q13049 receptors . In vivo tests , in turn , resulted in determining the functional activity of compounds 15 , 18 , 19 and 30 to the P08908 receptor . The results of these tests indicate that all of the ligands possess properties characteristic of P08908 receptor agonists . APRIL and Q9Y275 promote increased viability of replicating human B2 cells via mechanism involving cyclooxygenase 2 . Of relevance to both protective and pathogenic responses to Ag is the recent finding that soluble molecules of the innate immune system , i.e. , P05112 , B cell-activation factor of the P01375 family ( Q9Y275 ) , and P01024 , exhibit significant synergy in promoting the clonal expansion of human B2 cells following low-level P11274 ligation . Although P05112 , Q9Y275 , and C3dg each contribute to early cell cycle entry and progression to S phase , only Q9Y275 promotes later sustained viability of progeny needed for continued cycling . The present study sought to further clarify the mechanisms for Q9Y275 's multiple functions . By comparing Q9Y275 and a proliferation-inducing ligand ( APRIL ) efficacy at different stages in the response ( only Q9Y275 binds Q96RJ3 ; both bind transmembrane activator and calcium modulator and cyclophilin ligand interactor ( O14836 ) and B cell maturation Ag , the early role was attributed to Q96RJ3 , while the later role was attributed to O14836 /B cell maturation Ag . Importantly , Q9Y275 - and APRIL-promoted viability of cycling lymphoblasts was associated with sustained expression of cyclooxygenase 2 ( P35354 ) , the rate-limiting enzyme for DB00917 synthesis , within replicating cells . Supernatants of cultures with Q9Y275 and APRIL contained elevated DB00917 . Although P35354 inhibitors diminished daughter cell viability , exogenous DB00917 ( 1-1000 nM ) increased the viability and recovery of lymphoblasts . Increased yield of viable progeny was associated with elevated Mcl-1 , suggesting that a Q9Y275 /APRIL --> O14836 --> P35354 --> DB00917 --> Mcl-1 pathway reduces activation-related , mitochondrial apoptosis in replicating human B2 cell clones . Binding of antipsychotic drugs to human brain receptors focus on newer generation compounds . Using radioligand binding assays and post-mortem normal human brain tissue , we obtained equilibrium dissociation constants ( K(d)s ) for nine new antipsychotic drugs ( iloperidone , melperone , olanzapine , ORG 5222 , quetiapine , risperidone , sertindole , ziprasidone , and zotepine ) , one metabolite of a new drug ( 9-OH-risperidone ) , and three older antipsychotics ( clozapine , haloperidol , and pimozide ) at nine different receptors ( alpha1-adrenergic , alpha2-adrenergic , dopamine D2 , histamine H1 , muscarinic , and serotonin P08908 , P28221 , 5- Q13049 , and P28335 receptors ) . DB04946 was the most potent drug at the two adrenergic receptors . ORG 5222 was the most potent drug at dopamine D2 and 5-HT2c receptors , while ziprasidone was the most potent compound at three serotonergic receptors ( P08908 , P28221 , and 5- Q13049 ) . At the remaining two receptors , olanzapine was the most potent drug at the histamine H1 receptor ( Kd=0.087 nM ) ; clozapine at the muscarinic receptor ( Kd=9 nM ) . Certain therapeutic and adverse effects , as well as certain drug interactions can be predicted from a drug 's potency for blocking a specific receptor . These data can provide guidelines for the clinician in the choice of antipsychotic drug . The P11274 -ABL35INS insertion/truncation mutant is kinase-inactive and does not contribute to tyrosine kinase inhibitor resistance in chronic myeloid leukemia . Chronic myeloid leukemia is effectively treated with imatinib , but reactivation of P11274 - P00519 frequently occurs through acquisition of kinase domain mutations . The additional approved P00519 tyrosine kinase inhibitors ( TKIs ) nilotinib and dasatinib , along with investigational TKIs such as ponatinib ( DB08901 ) and P43146 -2036 , support the possibility that mutation-mediated resistance in chronic myeloid leukemia can be fully controlled ; however , the molecular events underlying resistance in patients lacking P11274 - P00519 point mutations are largely unknown . We previously reported on an insertion/truncation mutant , P11274 - P00519 (35INS) , in which structural integrity of the kinase domain is compromised and all P00519 sequence beyond the kinase domain is eliminated . Although we speculated that P11274 - P00519 (35INS) is kinase-inactive , recent reports propose this mutant contributes to P00519 TKI resistance . We present cell-based and biochemical evidence establishing that P11274 - P00519 (35INS) is kinase-inactive and does not contribute to TKI resistance , and we find that detection of P11274 - P00519 (35INS) does not consistently track with or explain resistance in clinical samples from chronic myeloid leukemia patients . DB01095 enhances the inhibitory effects of a selective AT1 receptor blocker , valsartan , on atherosclerosis . We investigated the effects of a P04035 inhibitor ( statin ) on the inhibitory effects of an angiotensin II type-1 receptor ( AT1 ) blocker on atherosclerosis and explored cellular mechanisms . We gave apolipoprotein E null mice a high-cholesterol diet for 10 weeks and measured atherosclerotic plaque area and lipid deposition . Neither 1 mg/kg per day of valsartan nor 3 mg/kg per day of fluvastatin had any effect on blood pressure or cholesterol concentration ; however , both drugs decreased plaque area and lipid deposition after 10 weeks . We then reduced the doses of both drugs to 0.1 mg/kg per day and 1 mg/kg per day , respectively . At these doses , neither drug had an effect on atherosclerotic lesions . When both drugs were combined at these doses , a significant reduction in atherosclerotic lesions was observed . Similar inhibitory effects of valsartan or fluvastatin on the expressions of nicotinamide-adenine dinucleotide/nicotinamide-adenine dinucleotide phosphate oxidase subunits P13498 and p47phox , production of superoxide anion , the expression of monocyte chemoattractant protein-1 , and intercellular adhesion molecule-1 expression were observed . These results suggest that concomitant AT1 receptor and cholesterol biosynthesis blockade , particularly when given concomitantly , blunts oxidative stress and inflammation independent of blood pressure or cholesterol-related effects . Activity of ponatinib against clinically-relevant DB05213 -resistant kinase domain mutants of P36888 -ITD . Secondary point mutations in the P36888 ( P36888 ) tyrosine kinase domain ( KD ) are common causes of acquired clinical resistance to the P36888 inhibitors DB05213 ( quizartinib ) and sorafenib . DB08901 ( DB08901 ) is a multikinase inhibitor with in vitro and clinical activity in tyrosine kinase inhibitor ( TKI ) -resistant chronic myeloid leukemia , irrespective of P11274 - P00519 KD mutation . DB08901 has demonstrated early clinical efficacy in chemotherapy-resistant acute myeloid leukemia ( AML ) patients with internal tandem duplication ( ITD ) mutations in P36888 . We assessed the in vitro activity of ponatinib against clinically relevant P36888 -ITD mutant isoforms that confer resistance to DB05213 or sorafenib . Substitution of the P36888 " gatekeeper " phenylalanine with leucine ( F691L ) conferred mild resistance to ponatinib , but substitutions at the P36888 activation loop ( AL ) residue D835 conferred a high degree of resistance . Saturation mutagenesis of P36888 -ITD exclusively identified P36888 AL mutations at positions D835 , D839 , and Y842 . The switch control inhibitor P43146 -2036 was similarly inactive against P36888 AL mutations . On the basis of its in vitro activity against P36888 TKI-resistant F691 substitutions , further clinical evaluation of ponatinib in TKI-naïve and select TKI-resistant P36888 -ITD+ AML patients is warranted . Alternative strategies will be required for patients with TKI-resistant P36888 -ITD D835 mutations . Akt inhibitors as an HIV-1 infected macrophage-specific anti-viral therapy . BACKGROUND : Unlike P01730 + T cells , HIV-1 infected macrophages exhibit extended life span even upon stress , consistent with their in vivo role as long-lived HIV-1 reservoirs . RESULTS : Here , we demonstrate that PI3K/Akt inhibitors , including clinically available Miltefosine , dramatically reduced HIV-1 production from long-living virus-infected macrophages . These PI3K/Akt inhibitors hyper-sensitize infected macrophages to extracellular stresses that they are normally exposed to , and eventually lead to cell death of infected macrophages without harming uninfected cells . Based on the data from these Akt inhibitors , we were able to further investigate how HIV-1 infection utilizes the PI3K/Akt pathway to establish the cytoprotective effect of HIV-1 infection , which extends the lifespan of infected macrophages , a key viral reservoir . First , we found that HIV-1 infection activates the well characterized pro-survival PI3K/Akt pathway in primary human macrophages , as reflected by decreased P60484 protein expression and increased Akt kinase activity . Interestingly , the expression of HIV-1 or SIV Tat is sufficient to mediate this cytoprotective effect , which is dependent on the basic domain of Tat - a region that has previously been shown to bind p53 . Next , we observed that this interaction appears to contribute to the downregulation of P60484 expression , since HIV-1 Tat was found to compete with P60484 for p53 binding ; this is known to result in p53 destabilization , with a consequent reduction in P60484 protein production . CONCLUSION : Since HIV-1 infected macrophages display highly elevated Akt activity , our results collectively show that PI3K/Akt inhibitors may be a novel therapy for interfering with the establishment of long-living HIV-1 infected reservoirs .

MH_train_37

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interacts_with DB01184?

multiple_choice

P51787 , Q9Y6J6 , and Na+-coupled solute transporters form reciprocally regulating complexes that affect neuronal excitability . Na(+)-coupled solute transport is crucial for the uptake of nutrients and metabolic precursors , such as myo-inositol , an important osmolyte and precursor for various cell signaling molecules . We found that various solute transporters and potassium channel subunits formed complexes and reciprocally regulated each other in vitro and in vivo . Global metabolite profiling revealed that mice lacking Q9Y6J6 , a K(+) channel β subunit , showed a reduction in myo-inositol concentration in cerebrospinal fluid ( P04141 ) but not in serum . Increased behavioral responsiveness to stress and seizure susceptibility in Kcne2(-/-) mice were alleviated by injections of myo-inositol . Suspecting a defect in myo-inositol transport , we found that Q9Y6J6 and P51787 , a voltage-gated potassium channel α subunit , colocalized and coimmunoprecipitated with P53794 , a Na(+)-coupled myo-inositol transporter , in the choroid plexus epithelium . Heterologous coexpression demonstrated that myo-inositol transport by P53794 was augmented by coexpression of P51787 but was inhibited by coexpression of both P51787 and Q9Y6J6 , which form a constitutively active , heteromeric K(+) channel . P53794 and the related transporter Q8WWX8 were also inhibited by a constitutively active mutant form of P51787 . The activities of P51787 and P51787 - Q9Y6J6 were augmented by P53794 and the glucose transporter P13866 but were suppressed by Q8WWX8 . Channel-transporter signaling complexes may be a widespread mechanism to facilitate solute transport and electrochemical crosstalk . LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Q9UBC3 interacts with O60264 chromatin remodeling enzyme , HDACs 1 and 2 , and components of the histone methylation system . The non-random pattern of genome-wide DNA methylation in mammalian cells is established and maintained by DNA methyltransferases P26358 , 3A , and 3B . De novo DNA methyltransferase Q9UBC3 is critical for embryonic development and is mutated in ICF syndrome . Despite its importance in normal cellular functioning , little is known about how Q9UBC3 operates in the context of chromatin . Here we demonstrate that Q9UBC3 associates with four chromatin-associated enzymatic activities common to transcriptionally repressed , heterochromatic regions of the genome : DNA methyltransferase , histone deacetylase , ATPase , and histone methylase activities . By immunoprecipitation and Q86UG4 pull-down , we show that Q9UBC3 interacts with Q13547 , Q92769 , P59665 proteins , Suv39h1 , and the DB00171 -dependent chromatin remodeling enzyme O60264 . Endogenous O60264 is also associated with DNA methyltransferase activity . These proteins co-localize extensively with Q9UBC3 in heterochromatic regions . Our results therefore link Q9UBC3 to three other components of the epigenetic machinery and provide important insights into how DNA methylation patterns may be established within the chromatin environment . DB01184 treatment for gastroparesis : demographic and pharmacogenetic characterization of clinical efficacy and side-effects . BACKGROUND : DB01184 is a useful alternative to metoclopramide for treatment of gastroparesis due to better tolerability . Effectiveness and side-effects from domperidone may be influenced by patient-related factors including polymorphisms in genes encoding drug-metabolizing enzymes , drug transporters , and domperidone targets . AIMS : The aim of this study was to determine if demographic and pharmacogenetic parameters of patients receiving domperidone are associated with response to treatment or side-effects . METHODS : Patients treated with domperidone for gastroparesis provided saliva samples from which DNA was extracted . Fourteen single-nucleotide polymorphisms ( SNPs ) in seven candidate genes ( P08183 , P10635 , P14416 , P15382 , Q9Y6J6 , Q12809 , P51787 ) were used for genotyping . SNP microarrays were used to assess single-nucleotide polymorphisms in the ADRA1A , P35368 , and P25100 loci . RESULTS : Forty-eight patients treated with domperidone participated in the study . DNA was successfully obtained from each patient . Age was associated with effectiveness of domperidone ( p=0.0088 ) . Genetic polymorphism in Q12809 was associated with effectiveness of domperidone ( p=0.041 ) . The efficacious dose was associated with polymorphism in P08183 gene ( p=0.0277 ) . The side-effects of domperidone were significantly associated with the SNPs in the promoter region of P25100 gene . CONCLUSIONS : Genetic characteristics associated with response to domperidone therapy included polymorphisms in the drug transporter gene P08183 , the potassium channel Q12809 gene , and α1D -- adrenoceptor P25100 gene . Age was associated with a beneficial response to domperidone . If verified in a larger population , this information might be used to help determine which patients with gastroparesis might respond to domperidone and avoid treatment in those who might develop side-effects . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal . The additive effect of neurotransmitter genes in pathological gambling . As access to gambling increases there is a corresponding increase in the frequency of addiction to gambling , known as pathological gambling . Studies have shown that a number of different neurotransmitters are affected in pathological gamblers and that genetic factors play a role . Polymorphisms at 31 different genes involved in dopamine , serotonin , norepinephrine , GABA and neurotransmitters were genotyped in 139 pathological gamblers and 139 age , race , and sex-matched controls . Multivariate regression analysis was used with the presence or absence of pathological gambling as the dependent variable , and the 31 coded genes as the independent variables . Fifteen genes were included in the regression equation . The most significant were the P14416 , P21917 , Q01959 , P17752 , P18825 , NMDA1 , and P49768 genes . The r(2) or fraction of the variance was less than 0.02 for most genes . Dopamine , serotonin , and norepinephrine genes contributed approximately equally to the risk for pathological gambling . These results indicate that genes influencing a range of brain functions play an additive role as risk factors for pathological gambling . Multi-gene profiles in specific individuals may be of assistance in choosing the appropriate treatment . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Cdk5 phosphorylates dopamine D2 receptor and attenuates downstream signaling . The dopamine D2 receptor ( P14416 ) is a key receptor that mediates dopamine-associated brain functions such as mood , reward , and emotion . P12004 -dependent kinase 5 ( Cdk5 ) is a proline-directed serine/threonine kinase whose function has been implicated in the brain reward circuit . In this study , we revealed that the serine 321 residue ( S321 ) in the third intracellular loop of P14416 ( D2i3 ) is a novel regulatory site of Cdk5 . Cdk5-dependent phosphorylation of S321 in the D2i3 was observed in in vitro and cell culture systems . We further observed that the phosphorylation of S321 impaired the agonist-stimulated surface expression of P14416 and decreased G protein coupling to P14416 . Moreover , the downstream DB02527 pathway was affected in the heterologous system and in primary neuronal cultures from p35 knockout embryos likely due to the reduced inhibitory activity of P14416 . These results indicate that Cdk5-mediated phosphorylation of S321 inhibits P14416 function , providing a novel regulatory mechanism for dopamine signaling . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Interaction of early environment , gender and genes of monoamine neurotransmission in the aetiology of depression in a large population-based Finnish birth cohort . Objectives Depression is a worldwide leading cause of morbidity and disability . Genetic studies have recently begun to elucidate its molecular aetiology . The authors investigated candidate genes of monoamine neurotransmission and early environmental risk factors for depressiveness in the genetically isolated population-based Northern Finland Birth Cohort 1966 ( 12 058 live births ) . Design The authors ascertained and subdivided the study sample ( n=5225 ) based on measures of early development and of social environment , and examined candidate genes of monoamine neurotransmission , many of which have shown prior evidence of a gene-environment interaction for affective disorders , namely P31645 , Q8IWU9 , P21964 , P21397 and the dopamine receptor genes P21728 - P21918 . Results and conclusion The authors observed no major genetic effects of the analysed variants on depressiveness . However , when measures of early development and of social environment were considered , some evidence of interaction was observed . Allelic variants of P21964 interacted with high early developmental risk ( p=0.005 for rs2239393 and p=0.02 for rs4680 ) so that the association with depression was detected only in individuals at high developmental risk group ( p=0.0046 and β=0.056 for rs5993883-rs2239393-rs4680 risk haplotype CGG including Val158 ) , particularly in males ( p=0.0053 and β=0.083 for the haplotype CGG ) . Rs4274224 from P14416 interacted with gender ( p=0.017 ) showing a significant association with depressiveness in males ( p=0.0006 and β=0.0023 ; p=0.00005 and β=0.069 for rs4648318-rs4274224 haplotype GG ) . The results support the role of genes of monoamine neurotransmission in the aetiology of depression conditional on environmental risk and sex , but not direct major effects of monoaminergic genes in this unselected population . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Absence of clinically important Q12809 channel blockade by three compounds that inhibit phosphodiesterase 5 -- sildenafil , tadalafil , and vardenafil . Compounds that inhibit phosphodiesterase 5 ( O76074 ) have been developed for the treatment of erectile dysfunction . Because men with erectile dysfunction frequently have comorbid cardiovascular disease , they may have limited cardiac repolarization reserve and be at risk of arrhythmia if treated with medications that prolong ventricular repolarization . The human ether-a-go-go related gene ( Q12809 ) channel is important for repolarization in human myocardium and is a common target for drugs that prolong the QT interval . We studied the ability of three compounds that inhibit O76074 -- sildenafil , tadalafil , and vardenafil -- to block the Q12809 channel . Using a whole cell variant of the patch-clamp method , the Q12809 current was measured in a stably transfected human embryonic kidney cell line expressing the Q12809 channel . The compounds produced dose-dependent reductions in Q12809 current amplitude over a concentration range of 0.1 to 100 microM . The IC50 values were 12.8 microM for vardenafil and 33.3 microM for sildenafil . Because the maximum soluble concentration of tadalafil ( 100 microM ) produced only a 50.9 % inhibition of the Q12809 current amplitude , the IC50 value for tadalafil could not be determined with the Hill equation . DB00820 had the weakest capacity to block the Q12809 channel , producing a 50.9 % blockade at the maximum soluble concentration ( 100 microM ) , compared with 86.2 % for vardenafil ( 100 microM ) and 75.2 % for sildenafil ( 100 microM ) . In conclusion , the concentrations of the O76074 inhibitors required to evoke a 50 % inhibition of the Q12809 current were well above reported therapeutic plasma concentrations of free and total compound . None of the three compounds was a potent blocker of the Q12809 channel . 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 . Activation of PPARgamma by rosiglitazone attenuates intestinal Cl- secretion . The thiazolidinedione ( TZD ) drugs rosiglitazone ( Ro ) and pioglitazone ( Po ) are PPARgamma agonists in widespread clinical use as insulin-sensitizing agents in Type 2 diabetes . On the basis of recent evidence implicating PPARgamma as a positive modulator of intestinal epithelial differentiation , we hypothesized that TZD drugs might attenuate intestinal secretory function . To evaluate this possibility , we examined the effects of Ro and Po on electrogenic Cl- secretion [ short-circuit current ( I(sc) ) ] in mouse intestinal segments and in cultured human intestinal epithelial cells ( HT29-Cl.19A ) . As hypothesized , oral administration of Ro ( 20 mg.kg(-1).day(-1) ) to mice for 8 days markedly reduced intestinal I(sc) responses to DB02527 (forskolin)- and Ca2+ (carbachol)-dependent stimuli . In these Ro-treated mice , cholera toxin-induced intestinal fluid accumulation was reduced 65 % . With continued Ro treatment , the I(sc) response to carbachol recovered significantly , whereas that to forskolin remained attenuated . Treatment of HT29 cells for 5 days with 10 muM Ro or Po in vitro brought about a similar hyposecretory state . In HT29 cells , the loss of DB02527 -dependent Cl- secretion was attributable to a reduced expression of P13569 Cl- channel , P51787 K+ channel , and Na-K-2Cl cotransporter-1 proteins . The transient loss of Ca2+-dependent Cl- secretion involved an impairment of basolateral Ca2+-stimulated K+ channel activity without a detectable loss of K(Ca)3.1 channel protein . Our results establish TZD drugs as important modulators of intestinal Cl- secretory function . An analysis of DNA methylation in human adipose tissue reveals differential modification of obesity genes before and after gastric bypass and weight loss . BACKGROUND : Environmental factors can influence obesity by epigenetic mechanisms . Adipose tissue plays a key role in obesity-related metabolic dysfunction , and gastric bypass provides a model to investigate obesity and weight loss in humans . RESULTS : Here , we investigate DNA methylation in adipose tissue from obese women before and after gastric bypass and significant weight loss . In total , 485,577 CpG sites were profiled in matched , before and after weight loss , subcutaneous and omental adipose tissue . A paired analysis revealed significant differential methylation in omental and subcutaneous adipose tissue . A greater proportion of CpGs are hypermethylated before weight loss and increased methylation is observed in the 3' untranslated region and gene bodies relative to promoter regions . Differential methylation is found within genes associated with obesity , epigenetic regulation and development , such as P11597 , O15409 , P56524 , Q9UBC3 , P51787 and HOX clusters . We identify robust correlations between changes in methylation and clinical trait , including associations between fasting glucose and P56524 , Q8NCC5 and Q8IV53 in subcutaneous adipose . Genes investigated with differential promoter methylation all show significantly different levels of mRNA before and after gastric bypass . CONCLUSIONS : This is the first study reporting global DNA methylation profiling of adipose tissue before and after gastric bypass and associated weight loss . It provides a strong basis for future work and offers additional evidence for the role of DNA methylation of adipose tissue in obesity . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes .

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interacts_with DB09079?

multiple_choice

Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 , IL-1 receptor antagonist , P05112 , P05231 , macrophage inflammatory protein-1alpha , macrophage inflammatory protein-1beta , monocyte chemoattractant protein-1 ( P13500 ) , interferon-inducible protein 10 , and P15692 . In vitro , oxytocin had no impact on LPS effects in releasing P01375 , P05231 , and P13500 in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 levels . Association between type 2 diabetes genetic susceptibility loci and visceral and subcutaneous fat area as determined by computed tomography . Visceral fat accumulation has an important role in the development of several metabolic disorders , such as type 2 diabetes , dyslipidemia and hypertension . New genetic loci that contribute to the development of type 2 diabetes have been identified by genome-wide association studies . To examine the association of type 2 diabetes susceptibility loci and visceral fat accumulation , we genotyped 1279 Japanese subjects ( 556 men and 723 women ) , who underwent computed tomography for measurements of visceral fat area ( VFA ) and subcutaneous fat area ( SFA ) for the following single-nucleotide polymorphisms ( SNPs ) : Q04721 rs10923931 , Q6YHU6 rs7578597 , P37231 rs1801282 , Q9P2N4 rs4607103 , Q9Y6M1 rs1470579 , P15692 rs9472138 , Q86VZ6 rs864745 , CDKN2A/ P42772 rs564398 and rs10811661 , Q03014 rs1111875 and rs5015480 , Q9NQB0 rs7901695 , P51787 rs2237892 , Q14654 rs5215 and rs5219 , Q93063 rs1113132 , rs11037909 , and rs3740878 , P49286 rs10830963 , P81605 rs1153188 , P19075 / O75473 rs7961581 , and Q9C0B1 rs8050136 and rs9939609 . None of the above SNPs were significantly associated with VFA . The Q9C0B1 rs8050136 and rs9939609 risk alleles exhibited significant associations with body mass index ( BMI ; P=0.00088 and P=0.0010 , respectively ) and SFA ( P=0.00013 and P=0.00017 , respectively ) . No other SNPs were significantly associated with BMI or SFA . Our results suggest that two SNPs in the Q9C0B1 gene are associated with subcutaneous fat accumulation . The contributions of other SNPs are inconclusive because of a limitation of the sample power . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . DB09079 , a triple angiokinase inhibitor , enhances cytotoxic therapy response in pancreatic cancer . Angiogenesis remains a sensible target for pancreatic ductal adenocarcinoma ( PDAC ) therapy . P15692 , PDGF , FGF and their receptors are expressed at high levels and correlate with poor prognosis in human PDAC . DB09079 is a triple angiokinase inhibitor that targets P17948 /2/3 , P11362 /2/3 and PDGFRα/β signaling . We investigated the antitumor activity of nintedanib alone or in combination with the cytotoxic agent gemcitabine in experimental PDAC . DB09079 inhibited proliferation of cells from multiple lineages found in PDAC , with gemcitabine enhancing inhibitory effects . DB09079 blocked PI3K/MAPK activity and induced apoptosis in vitro and in vivo . In a heterotopic model , net local tumor growth compared to controls ( 100 % ) was 60.8 ± 10.5 % in the gemcitabine group , -2.1 ± 9.9 % after nintedanib therapy and -12.4 ± 16 % after gemcitabine plus nintedanib therapy . Effects of therapy on intratumoral proliferation , microvessel density and apoptosis corresponded with tumor growth inhibition data . In a PDAC survival model , median animal survival after gemcitabine , nintedanib and gemcitabine plus nintedanib was 25 , 31 and 38 days , respectively , compared to 16 days in controls . The strong antitumor activity of nintedanib in experimental PDAC supports the potential of nintedanib-controlled mechanisms as targets for improved clinical PDAC therapy . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion . DB01076 inhibits inflammatory angiogenesis in mice through down regulation of P15692 , P01375 and TGF-beta1 . While compelling evidence indicates beneficial effects of statins on inflammatory processes , besides their cholesterol-lowering activities , the actions on angiogenesis are less clear-cut . Our aim was to investigate the effects of atorvastatin on key components of inflammatory angiogenesis in the murine sponge model . Polyester-polyurethane sponges , used as a framework for fibrovascular tissue growth , were implanted in Swiss mice . DB01076 ( 0.6 , 3 mg/kg/day ) was given orally for 8 days in drinking water . The implants collected at day 9 postimplantation were processed for the assessment of hemoglobin , myeloperoxidase ( P05164 ) , N-acetylglucosaminidase ( NAG ) and collagen . Relevant inflammatory , angiogenic and fibrogenic cytokines were also determined . DB01076 treatment resulted in significant decrease in sponge vascularization ( Hb content ) and in P15692 levels at both doses . Neutrophil influx ( P05164 activity ) was not affected by the compound whereas macrophage recruitment ( NAG activity ) was inhibited , suggesting a degree of selectivity by atorvastatin for this cell population . The level of P13500 ( MCP1-JE ) was decreased only with 0.6 mg/kg . DB01076 was also able to reduce collagen deposition and the levels of transforming growth factor ( TGF-beta1 ) intraimplant , dose-dependently . The inhibitory function of atorvastatin on multiple parameters of main components of inflammatory angiogenesis revealed in this study is clearly associated with the modulatory effects of P04035 on P15692 , P01375 and TGF-beta1 production . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Ablation of cholesterol biosynthesis in neural stem cells increases their P15692 expression and angiogenesis but causes neuron apoptosis . Although sufficient cholesterol supply is known to be crucial for neurons in the developing mammalian brain , the cholesterol requirement of neural stem and progenitor cells in the embryonic central nervous system has not been addressed . Here we have conditionally ablated the activity of squalene synthase ( P37268 ) , a key enzyme for endogenous cholesterol production , in the neural stem and progenitor cells of the ventricular zone ( VZ ) of the embryonic mouse brain . Mutant embryos exhibited a reduced brain size due to the atrophy of the neuronal layers , and died at birth . Analyses of the E11.5-E15.5 dorsal telencephalon and diencephalon revealed that this atrophy was due to massive apoptosis of newborn neurons , implying that this progeny of the P37268 -ablated neural stem and progenitor cells was dependent on endogenous cholesterol biosynthesis for survival . Interestingly , the neural stem and progenitor cells of the VZ , the primary target of P37268 inactivation , did not undergo significant apoptosis . Instead , vascular endothelial growth factor ( P15692 ) expression in these cells was strongly upregulated via a hypoxia-inducible factor-1-independent pathway , and angiogenesis in the VZ was increased . Consistent with an increased supply of lipoproteins to these cells , the level of lipid droplets containing triacylglycerides with unsaturated fatty acyl chains was found to be elevated . Our study establishes a direct link between intracellular cholesterol levels , P15692 expression , and angiogenesis . Moreover , our data reveal a hitherto unknown compensatory process by which the neural stem and progenitor cells of the developing mammalian brain evade the detrimental consequences of impaired endogenous cholesterol biosynthesis .

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interacts_with DB01269?

multiple_choice

Making sense of anti- P00533 plus oxaliplatin-based therapy in the first-line treatment of metastatic colorectal cancer . P01133 receptor ( P00533 ) targeting in patients with wild-type P01116 metastatic colorectal cancer has been associated with clinical activity in first- , second- and third-line therapies . DB01269 , a human monoclonal antibody that targets EGFRs , had previously been associated with tumor regressions and improved progression-free survival in patients with chemotherapy-resistant metastatic colorectal cancer . Douillard et al. have reported on the results of the Phase III DB01269 Randomized Trial in Combination with Chemotherapy for Metastatic Colorectal Cancer to Determine Efficacy ( PRIME ) Phase III of fluoroucil , leucovorin and oxaliplatin , with and without panitumumab , in patients with metastatic colorectal cancer . This article focuses on the PRIME study and its implications on clinical practice . In addition , the results of the PRIME study in the context of another P00533 -targeting agent , cetuximab , when combined with oxaliplatin-based first-line treatment of metastatic colorectal cancer , is discussed . Autocrine signaling via release of DB00171 and activation of Q99572 receptor influences motile activity of human lung cancer cells . Extracellular nucleotides , such as DB00171 , are released from cells and play roles in various physiological and pathological processes through activation of P2 receptors . Here , we show that autocrine signaling through release of DB00171 and activation of Q99572 receptor influences migration of human lung cancer cells . Release of DB00171 was induced by stimulation with TGF-β1 , which is a potent inducer of cell migration , in human lung cancer H292 cells , but not in noncancerous BEAS-2B cells . Treatment of H292 cells with a specific antagonist of Q99572 receptor resulted in suppression of TGF-β1-induced migration . PC-9 human lung cancer cells released a large amount of DB00171 under standard cell culture conditions , and Q99572 receptor-dependent dye uptake was observed even in the absence of exogenous ligand , suggesting constitutive activation of Q99572 receptor in this cell line . PC-9 cells showed high motile activity , which was inhibited by treatment with ecto-nucleotidase and Q99572 receptor antagonists , whereas a Q99572 receptor agonist enhanced migration . PC-9 cells also harbor a constitutively active mutation in epidermal growth factor receptor ( P00533 ) . Treatment with P00533 tyrosine kinase inhibitor AG1478 suppressed both cell migration and Q99572 receptor expression in PC-9 cells . Compared to control PC-9 cells , cells treated with Q99572 antagonist exhibited broadened lamellipodia around the cell periphery , while AG1478-treated cells lacked lamellipodia . These results indicate that Q99572 -mediated signaling and P00533 signaling may regulate migration of PC-9 cells through distinct mechanisms . We propose that autocrine DB00171 - Q99572 signaling is involved in migration of human lung cancer cells through regulation of actin cytoskeleton rearrangement . Growth of V79 cells as xenograft tumors promotes multicellular resistance but does not increase spontaneous or radiation-induced mutant frequency . A Chinese hamster V79 xenograft model was developed to determine whether cells subjected to a hypoxic tumor microenvironment would be more likely to undergo mutation at the P00492 locus . V79-171b cells stably transfected with P15692 and EGFP were grown subcutaneously in immunodeficient NOD/ SCID mice . V79-VE tumors were characterized for host cell infiltration , doubling time , hypoxic fraction , vascular perfusion , and response to ionizing radiation . When irradiated in vitro , the mutant frequency for a given surviving fraction did not differ for cells grown in vivo or in vitro . Similar results were obtained using HCT116 human colorectal carcinoma cells grown as xenografts . However , V79-VE cells grown as xenografts were significantly more resistant to killing than monolayers . The background mutant frequency and the radiation-induced mutant frequency did not differ for tumor cells close to or distant from blood vessels . Similarly , tumor cells from well-perfused regions showed the same rate of strand break rejoining and the same rate of loss of phosphorylated histone P16104 as cells sorted from poorly perfused regions . Therefore , deleterious effects of the tumor microenvironment on DNA repair efficiency or mutation induction could not be demonstrated in these tumors . Rather , development of multicellular resistance in V79-VE tumors acted to reduce mutant frequency for a given dose of radiation . Early DB09150 /PET scanning as a pharmacodynamic marker of anti- P00533 antibody activity in colorectal cancer . DB01269 is an anti- P01133 receptor ( P00533 ) antibody approved for use in treatment of chemotherapy-refractory colorectal cancers lacking K- DB01367 mutations . Despite overall response rates approximating 10 % , no marker predictive of clinical benefit has been identified . We describe a chemotherapy-refractory patient whose clinical condition necessitated rapid identification of an effective agent in whom we used (18)F- DB09150 -positron emission tomography ( DB09150 -PET ) /computed tomographic scanning 48 hours after an initial dose of panitumumab to document a pharmacodynamic response to the antibody . The initial 46 % ± 2.7 % drop in SUV(max) of four target lesions correlated with a partial response by Response Evaluation Criteria in Solid Tumors and a > 90 % drop in serum carcinoembryonic antigen at 8 weeks , indicating that an early decrease in DB09150 uptake may predict subsequent clinical benefit in response to anti- P00533 antibody therapy in colorectal cancer . Convergent and divergent cellular responses by ErbB4 isoforms in mammary epithelial cells . Associations of ErbB4 ( Q15303 / Q15303 ) , the fourth member of the P00533 family , with cancer are variable , possibly as a result of structural diversity of this receptor . There are multiple structural isoforms of Q15303 arising by alternative mRNA splicing , and a subset undergo proteolysis that releases membrane-anchored and soluble isoforms that associate with transcription factors and coregulators to modulate transcription . To compare the differential and common signaling activities of full-length ( FL ) and soluble intracellular isoforms of Q15303 , four JM-a isoforms ( FL and soluble intracellular domain ( ICD ) CYT-1 and CYT-2 ) were expressed in isogenic MCF10A cells and their biologic activities were analyzed . Both FL and ICD CYT-2 promoted cell proliferation and invasion , and CYT-1 suppressed cell growth . Transcriptional profiling revealed several new and underexplored Q15303 -regulated transcripts , including : proteases/protease inhibitors ( P08254 and P07093 ) , the YAP/Hippo pathway ( P29279 , O00622 , and P09486 ) , the mevalonate/cholesterol pathway ( P04035 , Q01581 , P01130 , and Q9UBM7 ) , and cytokines ( P10145 , P78556 , and P09341 ) . Many of these transcripts were subsequently validated in a luminal breast cancer cell line that normally expresses Q15303 . Furthermore , ChIP-seq experiments identified O75689 , P02649 , P09486 , P16949 , and Q05195 as novel molecular targets of Q15303 . These findings clarify the diverse biologic activities of Q15303 isoforms , and reveal new and divergent functions . IMPLICATIONS : ErbB4 as a regulator of Hippo and mevalonate pathways provides new insight into milk production and anabolic processes in normal mammary epithelia and cancer . Therapeutic potential of DB01269 : a fully human anti-epidermal growth factor receptor monoclonal antibody for cancer treatment . Overexpression of the epidermal growth factor receptor ( P00533 ) has been observed in a wide variety of human cancers and is associated with a poor clinical prognosis . In many cases , growth of the tumor cells is dependent on P00533 -mediated signals , because inhibition of binding of factors to the P00533 leads to cell death . Using XenoMouse technology , a fully human P00533 -specific monoclonal antibody , DB01269 , with high affinity ( 5 x 10(-11) mol/L ) has been generated . DB01269 blocks binding of both epidermal growth factor and transforming growth factor alpha to the P00533 , inhibits tyrosine phosphorylation of the P00533 , and inhibits cellular proliferation . In vivo , DB01269 not only blocks formation of human epidermoid carcinoma A431 xenografts in athymic mice , but also mediates therapeutic elimination of established tumors and acts cooperatively with chemotherapeutics in mediating tumor regression . These observations provide a strong basis for the development of DB01269 as a therapeutic agent for human solid tumors that overexpress P00533 . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Targeted agents for the treatment of advanced renal cell carcinoma . Renal cell carcinoma ( RCC ) is a highly treatment-resistant tumor type ; however , advances in elucidating the molecular pathophysiology underlying RCC has led to the identification of promising targets for therapeutic intervention . In clear-cell RCC , mutations to the von Hippel-Lindau ( P40337 ) gene results in the up regulation of many proteins necessary for tumor growth and survival -- such as vascular endothelial growth factor ( P15692 ) , basic fibroblast growth factor ( P09038 ) and platelet derived growth factor ( PDGF ) , which are involved in tumor-initiated angiogenesis . Q16790 and signaling via the epidermal growth factor receptor ( P00533 ) are involved in tumor cell proliferation and are also up regulated by mutation in the P40337 gene . The intracellular messenger pathways phosphoinositide 3-kinase ( PI3K ) and Raf/MEK/ P29323 act as convergence points for positive growth signaling ; the Raf/MEK/ P29323 pathway is also implicated in apoptosis . Several agents in development target P15692 ( bevacizumab ) , the P15692 receptor ( PTK787 , SU11248 , P15692 -trap , and BAY 43-9006 ) , the PDGF receptor ( SU11248 and BAY 43-9006 ) , or the P01133 receptor ( gefitinib , cetuximab , DB01269 , and erlotinib ) . The intracellular Raf/MEK/ P29323 signaling cascade has been targeted at either the level of Raf ( BAY 43-9006 , ISIS 5132 ) or MEK ( CI-1040 , PD184352 and ARRY-142886 ) , and PI3K signaling is disrupted by CCI-779 . DB05304 targets the Q16790 antigen , and PS-341 disrupts the 26S proteasome mediating the degradation of intracellular proteins . Given that multiple pathways contribute to tumor growth , anti-tumor activity may be increased by agents targeting multiple pathways , or by combining agents to allow horizontal or vertical inhibition of multiple pathways . DB01269 : a new anti- P00533 antibody for the treatment of advanced colorectal cancer . Combined effects of C225 and 125-iodine seed radiation on colorectal cancer cells . BACKGROUND : To characterize the effect of combined treatment of the anti-epidermal growth factor receptor ( P00533 ) monoclonal antibody C225 and 125-iodine ( 125I ) seed radiation in human colorectal cancer . METHODS : We treated LS180 cells with 125I continuous low dose rate radiation in the presence and absence of 100 nM C225 . The clonogenic capacity , cellular proliferation , cell cycle distribution , apoptosis , and molecular pathways of the cells following the treatments were analyzed in vitro . RESULTS : The sensitizer enhancement ratio of C225 was approximately 1.4 . Treatment with C225 and radiation alone produced significant inhibition of cell growth , but combination therapy produced greater inhibition than either treatment administered alone . C225 increased the radiation-induced apoptosis and the fraction of γ- P16104 foci positive cells at 48 h after treatment . The Akt phosphorylation level was lower in the cells receiving the combination treatment than in the cells treated with radiation or C225 alone . CONCLUSIONS : These findings indicate that C225 sensitizes LS180 cells to 125I seed radiation . Growth inhibition is mediated by inducing apoptosis and not cell cycle arrest . Additionally , we confirmed that C225 impairs DNA repair by reducing the cellular level of the P78527 and P12956 proteins . Furthermore , the inhibition of Akt signaling activation may be responsible for the C225-mediated radiosensitization . P00533 as a therapeutic target in colorectal cancer . The epidermal growth factor receptor ( P00533 ) is widely expressed in advanced colorectal cancers ( CRCs ) , and higher levels of P00533 are inversely related to survival in these patients . Two general strategies have been used to block P00533 signaling : preventing ligand binding with anti- P00533 monoclonal antibodies ( eg , cetuximab and DB01269 ) and inhibiting its intrinsic tyrosine kinase with small molecules ( eg , gefitinib [ DB00317 ] and erlotinib [ DB00530 ,Tarceva ] ) . Phase II trials of cetuximab suggest that it might be an effective treatment option alone or in combination with standard therapies as first- or second-line therapy . Phase I studies evaluating other P00533 inhibitors in patients with CRC have been reported . The inclusion of anti- P00533 therapies into standard treatment is the subject of current clinical trials . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . DB01269 -induced interstitial lung disease in a case of metastatic colorectal cancer . BACKGROUND : A Japanese postmarketing survey of panitumumab revealed that panitumumab-associated interstitial lung disease ( ILD ) occurred in approximately 1 % ( 19/1767 ) of patients , causing death in 36.8 % of these cases . CASE REPORT : We report the case of a 60-year-old Japanese man who developed ILD associated with panitumumab therapy ( third-line therapy ) for metastatic sigmoid colon cancer involving the liver , lymph nodes , and lungs . 2 months after the initiation of panitumumab therapy , he developed a progressive nonproductive cough , dyspnea , and a fever , and was diagnosed with ILD . Intravenous pulse methylprednisolone treatment led to quick recovery . The patient had some risk factors for ILD associated with epidermal growth factor receptor ( P00533 ) -tyrosine kinase inhibitors . CONCLUSION : Further studies are required to elucidate the association between anti- P00533 antibodies and ILD . Current situation of DB01269 , DB05101 , Nimotuzumab and Zalutumumab . P00533 overexpression usually correlates with a more advanced disease stage , a poorer prognosis and a worse chemotherapy response . P00533 expression increase has been observed in many tumours . For all the aforementioned reasons , P00533 inhibition can be considered an attractive approach in cancer treatment . One strategy has been receptor inhibition of extracellular domain using monoclonal antibodies . Cetuximab is the most developed one and there is plenty information on the literature about its current status . In this review we focus on other P00533 monoclonal antibodies under clinical development . The more developed one is DB01269 . Its clinical development is taking place very quickly and it has mainly been studied in colorectal cancer showing promising results . There are also other interesting drugs such as DB05101 , Nimotuzumab and Zalutumumab . P01116 mutations : analytical considerations . Colorectal cancer ( CRC ) is the third most common cancer and the second most common cause of cancer death globally . Significant improvements in survival have been made in patients with metastasis by new therapies . For example , Cetuximab and DB01269 are monoclonal antibodies that inhibit the epidermal growth receptor ( P00533 ) . P01116 mutations in codon 12 and 13 are the recognized biomarkers that are analyzed in clinics before the administration of anti- P00533 therapy . Genetic analyses have revealed that mutations in P01116 predict a lack of response to DB01269 and Cetuximab in patients with metastatic CRC ( mCRC ) . Notably , it is estimated that 35-45 % of CRC patients harbor P01116 mutations . Therefore , P01116 mutation testing should be performed in all individuals with the advanced CRC in order to identify the patients who will not respond to the monoclonal P00533 antibody inhibitors . New techniques for P01116 testing have arisen rapidly , and each technique has advantages and disadvantages . Herein , we review the latest published literature specific to P01116 mutation testing techniques . Since reliability and feasibility are important issues in clinical analyses . Therefore , this review also summarizes the effectiveness and limitations of numerous P01116 mutation testing techniques . New targeted therapies in gastrointestinal cancers . Despite surgical , radiotherapeutic , and chemotherapeutic advances , a large proportion of gastrointestinal ( GI ) cancers remain incurable . An improved understanding of the molecular pathogenesis of cancer has promulgated the development of novel agents designed to target critical pathways involved in cancer development and progression . The crucial role of the epidermal growth factor receptor ( P00533 ) in tumor proliferation and the overexpression of P00533 in several GI cancers provides the rationale for targeting and interrupting this key signaling network . P00533 blockade through monoclonal antibodies ( C225 and DB01269 ) and tyrosine kinase inhibitors ( ZD1839 and DB00530 ) has translated into promising evidence of clinical benefit . Ras-mediated signal transduction has been targeted using inhibitors of farnesyl transferase ( R115777 and SCH66336 ) to block the post-translation modification of Ras . Inhibitors of vascular growth factor receptor ( bevacizumab and PTK787 ) and matrix metalloproteinase target the effects of the host environment . P35354 inhibitors in colorectal cancer and STI571 in GI stromal tumors represent novel therapies of interest for these specific GI cancers . Evidence suggests that novel agents can be administered alone or in combination with standard therapies with little additional toxicity . The results of ongoing and future research efforts will clarify the optimal use and survival benefit of targeted therapies for patients with GI malignancies . Overview of monoclonal antibodies and small molecules targeting the epidermal growth factor receptor pathway in colorectal cancer . The epidermal growth factor receptor ( P00533 ) provides survival signals and is overexpressed in the majority of colorectal cancers . As more is learned about the molecular details of P00533 signaling , antibodies can be designed to interfere with specific domains of the P00533 molecule . In this review , we analyze preclinical and current clinical data on P00533 -targeting molecules and their potential role in the treatment of colorectal cancer . Cetuximab binds to domain III of P00533 and hinders ligand binding . It is now approved by the US Food and Drug Administration for metastatic colorectal cancer treatment . DB01269 is another widely studied anti- P00533 antibody with similar properties . Bispecific antibodies are modified immunoglobulin molecules containing 2 different binding specificities . These antibodies can redirect the immune response against tumor cells by tethering effector cells such as CD3e-expressing T cells or CD16-expressing natural killer cells and granulocytes to the surface of cancer cells . Tyrosine kinase inhibitors are quinazoline-derived , low molecular weight synthetic molecules that can block the intracellular tyrosine kinase domain of several receptors , including P00533 , Erb2 , and vascular endothelial growth factor receptor , and thereby inhibit ligand-induced receptor phosphorylation and abrogate the biologic effect of P00533 signaling . The presence of skin rash and P00533 gene amplification have been advanced as possible predictors of clinical effectiveness of targeted anti- P00533 therapies . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Protein kinase Cdelta and calmodulin regulate epidermal growth factor receptor recycling from early endosomes through Arp2/3 complex and cortactin . The intracellular trafficking of the epidermal growth factor receptor ( P00533 ) is regulated by a cross-talk between calmodulin ( P62158 ) and protein kinase Cdelta ( PKCdelta ) . On inhibition of P62158 , PKCdelta promotes the formation of enlarged early endosomes and blocks P00533 recycling and degradation . Here , we show that PKCdelta impairs P00533 trafficking due to the formation of an F-actin coat surrounding early endosomes . The PKCdelta-induced polymerization of actin is orchestrated by the Arp2/3 complex and requires the interaction of cortactin with PKCdelta . Accordingly , inhibition of actin polymerization by using cytochalasin D or by overexpression of active cofilin , restored the normal morphology of the organelle and the recycling of P00533 . Similar results were obtained after down-regulation of cortactin and the sequestration of the Arp2/3 complex . Furthermore we demonstrate an interaction of cortactin with P62158 and PKCdelta , the latter being dependent on P62158 inhibition . In summary , this study provides the first evidence that P62158 and PKCdelta organize actin dynamics in the early endosomal compartment , thereby regulating the intracellular trafficking of P00533 . Skin toxicity evaluation protocol with panitumumab ( Q8NFP0 ) , a phase II , open-label , randomized trial evaluating the impact of a pre-Emptive Skin treatment regimen on skin toxicities and quality of life in patients with metastatic colorectal cancer . PURPOSE : DB01269 , a fully human monoclonal antibody targeting the epidermal growth factor receptor ( P00533 ) , is approved in the United States and Europe for the treatment of refractory metastatic colorectal cancer ( mCRC ) . Skin toxicities are the most common adverse events with P00533 inhibitors . This is the first study designed to examine differences between pre-emptive and reactive skin treatment for specific skin toxicities in patients with mCRC for any P00533 inhibitor . PATIENTS AND METHODS : Patients receiving panitumumab-containing therapy were randomly assigned 1:1 to pre-emptive or reactive treatment ( after skin toxicity developed ) . Pre-emptive treatment included use of skin moisturizers , sunscreen , topical steroid , and doxycycline . The primary end point of the study was the incidence of protocol-specified > or= grade 2 skin toxicities during the 6-week skin treatment period . Quality of life ( QOL ) was assessed with the Dermatology Life Quality Index ( DLQI ) . RESULTS : Of 95 enrolled patients , 48 received pre-emptive treatment , and 47 received reactive treatment . The incidence of protocol-specified > or= grade 2 skin toxicities during the 6-week skin treatment period was 29 % and 62 % for the pre-emptive and reactive groups , respectively . Mean DLQI score change from baseline to week 3 was 1.3 points and 4.2 points in the pre-emptive and reactive groups , respectively . CONCLUSION : The pre-emptive skin treatment regimen was well tolerated . The incidence of specific > or= grade 2 skin toxicities during the 6-week skin treatment period was reduced by more than 50 % in the pre-emptive group compared with the reactive group . Patients in the pre-emptive group reported less QOL impairment than patients in the reactive group . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . DB01269 : a new frontier of target therapy for the treatment of metastatic colorectal cancer . DB01269 is a fully human monoclonal IgG(2) antibody targeting the P01133 receptor ( P00533 ) . This agent represents a new class of drug owing to its fully human nature , and no need for premedication and loading dose . DB01269 selectively binds to P00533 , blocking the extracellular domain of the receptor , and has not been associated with the formation of any antibodies directed against it . The drug is indicated as monotherapy for the treatment of patients with P00533 -expressing metastatic colorectal carcinoma with non-mutated ( wild-type ) P01116 after failure of fluoropyrimidine- , oxaliplatin- and irinotecan-containing chemotherapy regimens . The safety profile is favorable and is generally well tolerated ; the most common toxicities are skin rashes and diarrhea . Therefore , panitumumab 's hypersensitivity reaction rate is lower when compared with a chimeric monoclonal antibody such as cetuximab . DB01269 increases the clinician 's repertoire of agents to treat metastatic colorectal carcinoma . The available clinical data are the most promising for a single-agent anti- P00533 monoclonal antibody in this disease at the present time . These new data open different clinical scenarios in metastatic colorectal carcinoma patients and encourage clinicians and basic researchers to investigate new therapeutic approaches for this patient subset . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Short PEG-linkers improve the performance of targeted , activatable monoclonal antibody-indocyanine green optical imaging probes . The ability to switch optical imaging probes from the quenched ( off ) to the active state ( on ) has greatly improved target to background ratios . The optimal activation efficiency of an optical probe depends on complete quenching before activation and complete dequenching after activation . For instance , monoclonal antibody-indocyanine green ( mAb-ICG ) conjugates , which are promising agents for clinical translation , are normally quenched , but can be activated when bound to a cell surface receptor and internalized . However , the small fraction of commonly used ICG derivative ( ICG-Sulfo-OSu ) can bind noncovalently to its mAb and is , thus , gradually released from the mAb leading to relatively high background signal especially in the liver and the abdomen . In this study , we re-engineered a mAb-ICG conjugate , ( DB01269 -ICG ) using bifunctional ICG derivatives ( ICG-PEG4-Sulfo-OSu and ICG-PEG8-Sulfo-OSu ) with short polyethylene glycol ( PEG ) linkers . Higher covalent binding ( 70-86 % ) was observed using the bifunctional ICG with short PEG linkers resulting in less in vivo noncovalent dissociation . DB01269 -ICG conjugates with short PEG linkers were able to detect human epidermal growth factor receptor 1 ( P00533 ) -positive tumors with high tumor-to-background ratios ( 15.8 and 6.9 for P00533 positive tumor-to-negative tumor and tumor-to-liver ratios , respectively , at 3 d postinjection ) . [ DB01269 -treatment of metastatic colorectal cancer ] . In the molecular target treatment strategy of metastatic colorectal cancer patients panitumumab represents a new class of drugs due to its fully human nature , and no need for premedication and loading dose . DB01269 binds to epidermal growth factor receptor ( P00533 ) selectively . In registration pivotal studies the analysis of patient subgroups for P01116 status gives strong evidence for the important role of DB01367 oncogene : median progression-free survival was 16 weeks on panitumumab arm in P01116 wild-type patients ( 8 weeks in best supportive care ) , while in P01116 mutant patients panitumumab showed no efficacy , however adverse events were more frequent and severe . According to SmPC , panitumumab is indicated as monotherapy for the treatment of patients with P00533 expressing metastatic colorectal carcinoma with non-mutated ( wild-type ) P01116 after failure of fluoropyrimidine- , oxaliplatin- , and irinotecan-containing chemotherapy regimens . Adverse events are similar as with other P00533 inhibitors : skin symptoms ( rash ) , lung infiltrates , diarrhoea , ion abnormalities of renal origin . The drug was formerly available via named patient reimbursement , and now is financed by diagnosis-related group ( Q86YR7 ) system . Treatment of metastatic colorectal cancer : focus on panitumumab . Targeted agents are an important therapeutic option in the treatment of metastatic colorectal cancer ( mCRC ) . DB01269 is a recombinant , fully humanized , immunoglobulin G2 monoclonal antibody that targets the epidermal growth factor receptor ( P00533 ) with efficacy in mCRC as monotherapy and in combination with chemotherapy . Kirsten rat sarcoma ( P01116 ) mutation status has emerged as an important biomarker to predict response to anti- P00533 therapy . Optimal timing for panitumumab use in the mCRC treatment algorithm has not been established . This review discusses the mechanism of action , predictive biomarkers , and role of panitumumab in the treatment of mCRC . A case report on efficacy of Abound™ for anti- P00533 antibody-associated skin disorder in metastatic colon cancer . BACKGROUND : DB01269 is a full human epidermal growth factor receptor ( P00533 ) monoclonal antibody , an agent for metastatic colorectal cancer therapy . One of the most general adverse events of anti- P00533 monoclonal antibody therapy is skin disorder . At the present time , although prophylaxis of skin disorder is important for continuation of cancer therapy , there are no effective precautionary treatments . CASE PRESENTATION : A 73-year-old male with sigmoid colon cancer and synchronous lung metastasis was treated with panitumumab , an alone anti- P00533 monoclonal antibody as the third-line therapy.During the nine courses of the therapy , the response was stable disease ( SD ) , but skin disorder gradually appeared obviously ( CTCAE version 4.0 : Grade 2 ) . After 1 month of administration of Abound™ , symptoms of the skin disorder improved ( CTCAE version 4.0 : Grade 1 ) , thus the antibody therapy could be continued . CONCLUSION : We report that Abound™ was apparently effective in the treatment for anti- P00533 antibody-associated skin disorder . In the future , Abound™ could be expected as an agent for skin disorder as one of the side effects of colorectal cancer therapy . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Effects of P28335 -receptor expression on cell proliferation control in hamster fibroblasts : serotonin fails to induce a transformed phenotype . 5-HT1c receptors have been shown to act as protooncogenes in NIH 3T3 cells , inducing ligand-dependent focus formation . In order to assess their mitogenic and oncogenic potential in a different cell system , we transfected these receptors into CCL39 hamster fibroblasts , a well-characterized growth factor-dependent cell line . Cell clones expressing functional receptors were isolated and tested for ( a ) growth factor dependence of proliferation measuring thymidine incorporation in response to varying doses of serum , ( b ) the response to serotonin alone or in combination with other growth factors , and ( c ) the capacity for anchorage-independent proliferation . In the absence or presence of serotonin , the large majority of the clones isolated showed normal morphology and normal growth factor dependence and was unable to grow in soft agar . None of the clones showed a significant response to serotonin alone in DNA synthesis reinitiation experiments , but synergy was observed between serotonin and the tyrosine kinase activating growth factors P01133 and FGF . However , the major part of this effect could be abolished by an antagonist of 5-HT1b receptors , which are endogenous in CCL39 cells . The same receptor was found to mediate a significant mitogenic response to the neurotransmitter in Ha-ras-transfected cells . The fact that 5-HT1c receptors do not readily induce a transformed phenotype in CCL39 cells clearly distinguishes them from strong dominantly acting oncogene products like DB01367 , P12931 , or P07333 . Study of P01116 new predictive marker in a clinical laboratory . BACKGROUND : The presence of somatic mutations in the P01116 gene has been identified as a reliable strong negative predictor for the response to targeting the epidermal growth factor receptor ( P00533 ) , in patients with metastatic colorectal cancer and the use of anti- P00533 monoclonal antibodies such as Cetuximab and DB01269 is now restricted to patients with no detectable P01116 mutations . Between 30 and 40 % of colorectal cancers contain a mutated P01116 oncogene . The aim of this study was to evaluate concordance between three methods to analyze P01116 mutational status in regard to clinical testing . METHODS : We analyzed P01116 mutations in codons 12 and 13 of exon 2 in one hundred formalin-fixed paraffin-embedded ( FFPE ) colorectal cancer samples by three different methods : Direct Sequencing and two commercial kits on allele-specific oligonucleotide hybridization ( P01116 StripAssay , Vienna Lab. ) and Amplification Refractory Mutation System/Scorpions ( Q9ULH0 /S ; TheraScreen P01116 Mutation kit DxS ) based on q-PCR . RESULTS : We have found similar frequencies of P01116 mutations by TheraScreen and Strip-Assay ( 44 and 48 % ) , with a κ value of 0.90 , indicating almost perfect agreement between methods . The frequency by direct sequencing was much lower ( 26 % ) and the κ values were 0.67 ( compared to TheraScreen ) and 0.57 ( compared to Strip-Assay ) indicating low sensitivity . CONCLUSIONS : On analyzing P01116 mutation in FFPE tumor samples , direct sequencing sensitivity is too low to be used in a clinical setting . Choosing between Q9ULH0 /S ; TheraScreen P01116 Mutation kit DxS and P01116 StripAssay , Vienna Lab , will depend on laboratory facilities and expertise . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Targeting the P00533 pathway for cancer therapy . Clinical studies have shown that HER-2/Neu is over-expressed in up to one-third of patients with a variety of cancers , including B-cell acute lymphoblastic leukemia ( B-ALL ) , breast cancer and lung cancer , and that these patients are frequently resistant to conventional chemo-therapies . Additionally , in most patients with multiple myeloma , the malignant cells over-express a number of epidermal growth factor receptors ( P00533 )s and their ligands , HB- P01133 and amphiregulin , thus this growth-factor family may be an important aspect in the patho-biology of this disease . These and other , related findings have provided the rationale for the targeting of the components of the P00533 signaling pathways for cancer therapy . Below we discuss various aspects of P00533 -targeted therapies mainly in hematologic malignancies , lung cancer and breast cancer . Beside novel therapeutic approaches , we also discuss specific side effects associated with the therapeutic inhibition of components of the P00533 -pathways . Alongside small inhibitors , such as DB01259 ( DB01259 , GW572016 ) , Gefitinib ( DB00317 , ZD1839 ) , and Erlotinib ( Tarceva , DB00530 ) , a significant part of the review is also dedicated to therapeutic antibodies ( e.g. : DB00072 /Herceptin , DB06366 / DB06366 /rhuMab-2C4 , Cetuximab/Erbitux/IMC-C225 , DB01269 /Abenix/ DB01269 , and also DB05294 ) . In addition , we summarize , both current therapy development driven by antibody-based targeting of the P00533 -dependent signaling pathways , and furthermore , we provide a background on the history and the development of therapeutic antibodies . DB01269 a novel drug in cancer treatment . The epidermal growth factor receptor ( P00533 ) is a member of the erbB family overexpressed in most of the solid tumors . In cancer cells , the overexpression of P00533 correlates with the development and the progression of tumor . DB01269 is a fully human monoclonal antibody that blocks the extracellular domain of the P00533 and has not been associated with the formation of any antibodies directed against it . This review summarizes on the preclinical and clinical development of panitumumab in human solid tumors . As bevacizumab and cetuximab have been approved for colorectal cancer because of their improvements in progression-free survival and overall survival when associated with chemotherapy , panitumumab represents an interesting molecule which needs more phase III studies to validate its efficacy . Multiplexed evaluation of a cell-based assay for the detection of antidrug neutralizing antibodies to panitumumab in human serum using automated fluorescent microscopy . The method described here employs a high-content cell-based assay format for the detection of neutralizing antibodies ( NAbs ) to panitumumab , a fully human monoclonal antagonistic antibody to the human epidermal growth factor ( P01133 ) receptor in human serum ( screening assay ) . A specificity assay was also developed and qualified to confirm that the neutralizing activity was attributable to the presence of NAbs and not due to serum interference ( serum interference assay ) . The ArrayScan IV HCS reader was used for the measurement of tyrosine phosphorylation of the epidermal growth factor receptor ( P00533 ) and P35610 -1 redistribution between the cytoplasm and nucleus in the human epidermoid carcinoma cell line A431 . Assay conditions were developed by ( 1 ) optimizing the response of the A431 cells to recombinant human P01133 in pooled human serum , ( 2 ) evaluating the ability of panitumumab to inhibit the P01133 response , and ( 3 ) assessing the assay 's sensitivity for detecting a positive control affinity purified rabbit polyclonal anti-panitumumab antibody . DB01269 dose-dependently inhibited 4 ng/mL P01133 , and the positive control antibody showed a dose-dependent neutralization of 50 ng/mL panitumumab . The experiments indicated that in comparison to P35610 -1 translocation , P00533 phosphorylation was the optimal endpoint for the screening and serum interference assays . Assay cut points were derived for the screening and serum interference assays by obtaining normalized ratios of mean fluorescence intensity values obtained with P00533 phosphorylation by testing sera from healthy human donor sera . The assay sensitivity was determined to be 0.125 microg/mL for the positive control antibody . Vascular Endothelial Growth Factor plus Epidermal Growth Factor Receptor Dual Targeted Therapy in Metastatic Colorectal Cancer : Synergy or Antagonism ? There has been an intensive effort to develop novel therapies for the treatment of metastatic colorectal cancer ( mCRC ) . The anti-epidermal growth factor receptor ( P00533 ) antibodies panitumumab and cetuximab and the anti-vascular endothelial growth factor ( P15692 ) antibody bevacizumab have demonstrated clinical efficacy and acceptable toxicity in the treatment of mCRC as single agents or in combination with chemotherapy . Recent clinical trials have explored the efficacy and safety of treatment regimens incorporating chemotherapy in combination with bevacizumab and either panitumumab or cetuximab in patients with mCRC . Results from the BOND-2 trial , which investigated cetuximab , bevacizumab , and chemotherapy in mCRC , provided support for this therapeutic approach . Two large randomized phase 3 trials were initiated to evaluate firstline treatment of mCRC . The DB01269 Advanced Colorectal Cancer Evaluation ( PACCE ) study investigated the efficacy and safety of oxaliplatin- or irinotecan-based chemotherapy and bevacizumab with or without panitumumab ; CAIRO2 assessed the efficacy and safety of capecitabine/oxaliplatin and bevacizumab with or without cetuximab . In both trials , the combination of bevacizumab , an P00533 -specific antibody , and chemotherapy in first-line treatment of mCRC was associated with increased toxicity and no improvement in patient outcome . These results suggest that these specific combinations should not be used in first-line mCRC outside investigational studies . Boca-dependent maturation of beta-propeller/ P01133 modules in low-density lipoprotein receptor proteins . The extracellular portions of cell surface receptor proteins are often comprised of independently folding protein domains . As they are translated into the endoplasmic reticulum ( ER ) , some of these domains require protein chaperones to assist in their folding . Members of the low-density lipoprotein receptor ( P01130 ) family require the chaperone called Boca in Drosophila or its ortholog , Mesoderm development , in the mouse . All LDLRs have at least one six-bladed beta-propeller domain , which is immediately followed by an epidermal growth factor ( P01133 ) repeat . We show here that Boca is specifically required for the maturation of these beta-propeller/ P01133 modules through the secretory pathway , but is not required for other P01130 domains . Protein interaction data suggest that as LDLRs are translated into the ER , Boca binds to the beta-propeller . Subsequently , once the P01133 repeat is translated , the beta-propeller/ P01133 module achieves a more mature state that has lower affinity for Boca . We also show that Boca-dependent beta-propeller/ P01133 modules are found not only throughout the P01130 family but also in the precursor to the mammalian P01133 ligand . Treatment with panitumumab after a severe infusion reaction to cetuximab in a patient with metastatic colorectal cancer : a case report . Monoclonal antibodies targeting the epidermal growth factor receptor ( P00533 ) are effective in the treatment of metastatic colorectal cancer . Cetuximab is a human-murine chimeric monoclonal antibody targeting the P00533 . Even with premedication , cetuximab can result in an infusion reaction in select patients . In a portion of these patients , the reaction is severe , and further therapy with cetuximab is contraindicated , thus preventing these patients from receiving potentially beneficial anti- P00533 therapy . DB01269 is a fully human monoclonal antibody also targeting the P00533 . DB01269 is given without premedication and , in clinical trials , has rarely been associated with infusion reactions . It is not yet known whether panitumumab can be safely given in patients with a previous severe reaction to cetuximab . We report a case of a patient successfully treated with panitumumab after the patient had a severe infusion reaction to cetuximab . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . DB01269 combined with irinotecan for patients with P01116 wild-type metastatic colorectal cancer refractory to standard chemotherapy : a GERCOR efficacy , tolerance , and translational molecular study . BACKGROUND : The purpose of this study was to evaluate the combination of panitumumab and irinotecan in patients with P01116 wild-type metastatic colorectal cancer refractory to standard chemotherapy ( oxaliplatin , fluoropyrimidines-irinotecan and bevacizumab ) . PATIENTS AND METHODS : P01116 status was first determined locally but subsequent validation of P01116 status and additional screenings ( rare P01116 , P01111 , P15056 mutations and P00533 copy number ) were centrally assessed . Patients received panitumumab ( 6 mg/kg ) and irinotecan ( 180 mg/m² ) every 2 weeks . RESULTS : Sixty-five eligible patients were analyzed . The objective response rate ( ORR ) was 29.2 % [ 95 % confidence interval ( 95 % CI ) 18.2-40.3 ] . Median progression-free and overall survivals were 5.5 and 9.7 months , respectively . Most frequent grade 3/4 toxic effects were skin 32.3 % , diarrhea 15.4 % and neutropenia 12.3 % . Tissue samples were available for 60 patients . For the confirmed P01116 wild-type population codon 12 or 13 mutation ( n = 54 ) , ORR was 35.2 % ( 95 % CI 22.4.1-47.9 ) . Thirteen patients had a P01111 , a P15056 or a rare P01116 mutation , and no tumor response was observed in this subgroup when compared with 46.3 % ( 95 % CI 31.1-61.6 ) ORR in the subgroup of 41 patients with no identified mutation . CONCLUSION : DB01269 and irinotecan is an active third-line regimen in a well-defined population based on biomarkers . ClinicalTrials.gov Identifier NCT00655499 . Safety and efficacy of panitumumab following cetuximab : retrospective review of the Memorial Sloan-Kettering experience . There are two highly selective antibodies to the epidermal growth factor receptor ( P00533 ) now available for use in metastatic colorectal cancer ( mCRC ) . In P01116 wild type patients , cetuximab ( Cmab ) -an IgG1 chimeric molecule -- has activity alone and in combination with chemotherapy for the first , second and third-line settings . DB01269 ( Pmab ) -- a fully humanized IgG2 molecule -- has activity as a single agent in chemorefractory mCRC and shows promising activity in combination with chemotherapy . It remains unclear which antibody to use . This retrospective review of our experience with Pmab in 13 P00533 antibody-naive patients and in 22 patients previously treated with Cmab for mCRC highlights a lack of hypersensitivity reactions ( HSR ) with Pmab , even in patients who experienced HSR to Cmab . In patients who received Cmab , 22 % developed grade 3-4 HSR . There were no HSR on subsequent treatment with Pmab . We demonstrate similar activity in 95 % of cases , between Cmab and Pmab both alone and in combination with chemotherapy in the treatment of mCRC . In one case we report unique sensitivity to Pmab after progression with Cmab . Monoclonal antibodies targeting the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 , P00533 ) autocrine pathway contributes to a number of highly relevant processes in cancer development and progression , including cell proliferation , regulation of apoptotic cell death , angiogenesis and metastatic spread . The crucial role that P00533 plays in human cancers has led to an extensive search for selective inhibitors of its signaling pathway . The results of a large body of preclinical studies and clinical trials thus far conducted suggest that targeting the P00533 could bring a significant contribution to cancer therapy . A variety of different approaches are currently being used to target the P00533 . The most promising strategies in clinical development include monoclonal antibodies , to prevent ligand binding , and small molecules inhibitors of the tyrosine kinase enzymatic activity , that inhibit autophosphorylation and downstream intracellular signaling . Several blocking monoclonal antibodies against the P00533 have been developed . Among these , IMC-225 is a chimeric human-mouse monoclonal IgG1 antibody that has been the first anti- P00533 targeted therapy to enter clinical evaluation in cancer patients in Phase II and III studies , alone or in combination with conventional radiotherapy and chemotherapy . However , other antibodies against P00533 have demonstrated antitumor activity in several preclinical models of human cancer and are currently under investigation in the clinical setting , such as ICR62 , DB01269 and EMD72000 . This review will focus on all the preclinical data available on monoclonal antibodies engineered against the P01133 receptor . Targeted agents for the treatment of advanced renal cell carcinoma . Metastatic renal cell carcinoma ( RCC ) is currently one of the most treatment-resistant malignancies . However , the elucidation of the molecular mechanisms underlying RCC development has led to the identification of promising targets for novel therapeutic agents . The involvement of the Von Hippel-Lindau protein pathway in clear cell RCC suggests that downstream targets of this pathway , namely , signaling through vascular endothelial growth factor ( P15692 ) in endothelial cells , platelet-derived growth factor ( PDGF ) in endothelial cells and pericytes , and the epidermal growth factor receptor ( P00533 ) pathway in tumor cells are all reasonable and rational therapeutic targets . A number of agents are in development that target P15692 ( bevacizumab , a recombinant , humanized monoclonal antibody ) or its receptor , VEGFR ( PTK787 , SU011248 , and BAY 43-9006 , all of which are small molecule inhibitors ) . Agents targeting P00533 also are being investigated clinically ( gefitinib , cetuximab , erlotinib , and DB01269 ) . The Raf/MEK/ P29323 pathway is an important downstream convergence point for signaling through VEGFR , platelet-derived growth factor receptor ( P09619 ) , and P00533 ( all have receptor tyrosine kinase activity ) and also has important antiapoptotic effects , thereby providing an attractive target for intervention . In addition to inhibiting VEGFR and P09619 -mediated angiogenic pathways , BAY 43-9006 has been shown to inhibit the Raf/MEK/ P29323 pathway at the level of Raf kinase . MEK-directed therapeutic approaches are also in development . Given that multiple molecular pathways are implicated in tumor cell growth , antitumor activity may be increased by using individual agents that target multiple pathways , or by combining different agents to allow vertical or horizontal inhibition of relevant pathways . Improved speciation characteristics of PEGylated indocyanine green-labeled DB01269 : revisiting the solution and spectroscopic properties of a near-infrared emitting anti- P00533 antibody for optical imaging of cancer . A water-soluble amine-reactive PEGylated analogue of near-infrared emitting dye indocyanine green ( 5 ) was synthesized and used to label the anti- P00533 antibody panitumumab ( Vectibix ) at various equivalents . These conjugates were compared with non-PEGylated analogue conjugate products and the solution speciation analyzed with UV-vis spectrophotometry , size exclusion HPLC , and SDS-PAGE . PEGylation of the bioconjugates was successful in preventing aggregation in solution , a phenomenon observed with the non-PEGylated bioconjugates presumably due to the hydrophobicity of indocyanine green . Competitive radioimmunoassay demonstrated that the targeting moiety of the PEGylated bioconjugates was conserved . Fluorescence microscopy also demonstrated membrane binding of the bioconjugate to P00533 -expressing A431 cells . Hence , these bioconjugates are suitable candidates for the in vivo optical imaging of P00533 -expressing tumors . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Development and characterization of 89Zr-labeled panitumumab for immuno-positron emission tomographic imaging of the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) is overexpressed in the majority of malignancies and has been associated with poor outcomes . DB01269 , an anti- P00533 monoclonal antibody that binds to the extracellular binding domain of P00533 , is increasingly used with radiotherapy and chemotherapy but has associated toxicities . The purpose of this study was to develop and characterize a novel targeted imaging agent for the P00533 using radiolabeled panitumumab . Flow cytometry studies were performed to evaluate P00533 expression in several cell lines . DB00746 -Bz-NCS ( DB00746 ) was conjugated to panitumumab and labeled with (89)Zr . Cell uptake studies were performed in four cell lines . For biodistribution studies and micro-positron emission tomography/computed tomography ( PET/CT ) , mouse xenograft models were generated using the same cell lines . PET was performed , and tumors and select organs were harvested for biodistribution studies . DB01269 was radiolabeled with (89)Zr with high radiochemical purity and specific activity and was found to be stable in serum . Cell binding studies demonstrated that radiotracer uptake in cells correlated with the degree of P00533 expression . MicroPET/CT imaging studies demonstrated a high intensity of (89)Zr-panitumumab in A431 and HCT 116 tumors in comparison with the P00533 -negative tumors . Biodistribution studies confirmed the results from the imaging studies . (89)Zr-panitumumab imaging of P00533 -positive tumors demonstrated levels of radiotracer uptake associated with P00533 expression . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . P00533 gene expression in avian epiphyseal growth-plate cartilage cells : effect of serum , parathyroid hormone and atrial natriuretic peptide . Avian chondrocytes and fibroblasts , derived from epiphyseal growth-plate and skin , respectively , were cultured in vitro . In chondrocytes , epidermal growth factor ( P01133 ) caused a dose-dependent stimulation of proliferation . P01133 receptor mRNA was not detected with the v-erb B probe in chondrocytes cultured in the presence of 5 % fetal calf serum ( FCS ) . In the absence of FCS in the medium , a time-dependent increase in the level of P01133 receptor mRNA was observed . Parallel changes were also observed in the level of P01133 receptor , as demonstrated by immunofluorescence using antibodies directed against avian P01133 receptor . In avian fibroblasts , P01133 receptor mRNA and P01133 receptor levels were not affected by FCS . Furthermore , FCS did not affect the level of thyroid hormone receptor mRNA ( using v-erb A as a probe ) in either chondrocytes or fibroblasts . Parathyroid hormone ( PTH ) , which acts as a mitogen in avian chondrocytes attenuated -- whereas atrial natriuretic peptide ( P01160 ) , a suppressor of chondrocyte proliferation , enhanced -- P01133 receptor mRNA . The present results show that avian growth-plate chondrocytes respond to P01133 and bear P01133 receptors . The levels of P01133 mRNA and P01133 receptor are inversely related to cell proliferation . The results also support previous suggestions that PTH and P01160 play important roles in chondrocyte proliferation , possibly through their effect on the synthesis of the P01133 receptor . DB01269 : leading to better overall survival in metastatic colorectal cancer ? INTRODUCTION : Survival of metastatic colorectal cancer ( mCRC ) patients has improved greatly over the past few years , essentially due to the appearance of new biological therapies . Among these new therapies , monoclonal antibodies targeting the P00533 are the leading contributors to the so-called personalized medicine . Biomarkers within the P00533 pathway , such as K-Ras mutation , have proved to help better select the patients benefiting from these treatments . AREAS COVERED : DB01269 is a fully human monoclonal antibody that targets the P00533 and is currently approved in combination with chemotherapy or in monotherapy for the treatment of mCRC patients . Following a description of the pharmacological and tolerability data , this review focuses on the clinical activity of panitumumab through the description of clinical trials and biomarker research . EXPERT OPINION : Recent biomarker research with expanded Ras testing has led to an improvement in overall survival for all Ras wild-type patients treated with panitumumab . Furthermore , the thorough evaluation of markers within the P00533 pathway could potentially prevent detrimental effects for patients treated with panitumumab and avoid unnecessary toxicity and costs . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . Celecoxib blocks cardiac Kv1.5 , Kv4.3 and Kv7.1 ( P51787 ) channels : effects on cardiac action potentials . Celecoxib is a P35354 inhibitor that has been related to an increased cardiovascular risk and that exerts several actions on different targets . The aim of this study was to analyze the effects of this drug on human cardiac voltage-gated potassium channels ( Kv ) involved on cardiac repolarization Kv1.5 ( I(Kur) ) , Kv4.3+ Q9NS61 ( I(to1) ) and Kv7.1+ P15382 ( I(Ks) ) and to compare with another P35354 inhibitor , rofecoxib . Currents were recorded in transfected mammalian cells by whole-cell patch-clamp . Celecoxib blocked all the Kv channels analyzed and rofecoxib was always less potent , except on Kv4.3+ Q9NS61 channels . Kv1.5 block increased in the voltage range of channel activation , decreasing at potentials positive to 0 mV . The drug modified the activation curve of the channels that became biphasic . Block was frequency-dependent , increasing at fastest frequencies . Celecoxib effects were not altered by DB08837 (out) in R487Y mutant Kv1.5 channels but the kinetics of block were slower and the degree of block was smaller with DB08837 (in) , indicating that celecoxib acts from the cytosolic side . We confirmed the blocking properties of celecoxib on native Kv currents from rat vascular cells , where Kv1.5 are the main contributors ( IC(50)≈ 7 μM ) . Finally , we demonstrate that celecoxib prolongs the action potential duration in mouse cardiac myocytes and shortens it in guinea pig cardiac myocytes , suggesting that Kv block induced by celecoxib may be of clinical relevance . Therapeutic integration of new molecule-targeted therapies with radiotherapy in lung cancer . Lung cancer is the most common form of the disease and the leading cause of cancer deaths worldwide . Non-small-cell lung cancer ( NSCLC ) accounts for approximately 80-85 % of all lung cancers . Forty percent of all cases present with stage III , and many of them are considered inoperable ( staged IIIA with mediastinal lymph node involvement ) or stage IIIB disease . Concurrent platinum-based chemotherapy and thoracic radiation has demonstrated survival benefits in these patients . We review the role of new target agents in combination with radiotherapy in stage III NSCLC . Antiangiogenics improve tumor oxygenation thereby improving the therapeutic efficacy of irradiation in models . DB00112 in combination with thoracic radiation has shown high toxicity . However , other antiangiogenic agents are more promising . Radiation activates epidermal growth factor receptor ( P00533 ) pathways , inducing radioresistance , cell proliferation and enhanced DNA repair . After promising data from preclinical models and early clinical trials , cetuximab did not show any benefit in a recent phase III trial . DB01269 and nimotuzumab are under evaluation . Gefitinib has been investigated in combination with radiotherapy for unresectable stage III NSCLC , but results in maintenance treatment after chemoradiotherapy were not encouraging . Erlotinib has also been tested in a phase II trial with chemoradiotherapy . Other new pathways and agents are being studied , such as m-TOR pathway , bortezomib , heat shock protein 90 ( Hsp90 ) inhibition , histone deacetylase inhibitors ( HDACS ) , aurora kinases , mitogen activated protein kinases ( Q9P0L2 ) and PARP inhibitors . An P01133 receptor inhibitor induces P10826 expression in breast and ovarian cancer cells . Inhibition of the epidermal growth factor ( P01133 ) -receptor ( P00533 ) has become a promising anticancer treatment strategy . In addition , application of retinoids yields encouraging results for cancer prevention and therapy . Many tumors express no or low amounts of retinoic acid receptor-beta2 ( RAR-beta2 ) due to epigenetic silencing via DNA hypermethylation . RAR-beta2 is the main mediator of the antiproliferative effect of retinoids . RAR-beta2 re-expression causes reversal of transformation , cell cycle arrest , and restoration of retinoid sensitivity . RAR-beta2 is thus a tumor suppressor . Western blotting , colorimetric in vitro cell proliferation assays , and reverse transcription-polymerase chain reaction demonstrated that the P00533 inhibitor PD153035 not only blocked activation of P00533 and inhibited cell growth , but also stimulated P10826 expression in MDA-MB-468 breast and OVCAR-3 ovarian carcinoma cells . Upregulation of P10826 by PD153035 was confirmed by real-time reverse transcription-polymerase chain reaction . In contrast , expression of other retinoid receptors and of estrogen receptor-alpha was not affected . PD153035-mediated re-induction of P10826 was associated with demethylation of the RAR-beta2 gene promoter P2 as demonstrated by methylation-specific polymerase chain reaction . These novel results on the ErbB/retinoid receptor cross-talk may be useful for designing future anticancer combination regimens . P40763 signaling is induced by intercellular adhesion in squamous cell carcinoma cells . The signal transducer and activator of transcription-3 ( P40763 ) frequently activated during tumor progression has been linked to enhanced cell growth . In squamous cell carcinoma of the head and neck ( HNSCC ) , P40763 signaling has been shown to inhibit apoptosis and induce a more aggressive phenotype through the activation of specific signaling pathways . In the present study , we have examined the potential mechanism by which cell-cell contact initiates P40763 activation . Using a panel of HNSCC cell lines , Ca(+2)-dependent cell-cell adhesion and adherens junction formation in multicellular aggregates triggered phosphorylation of P40763 -Y705 and P42224 -Y701 . This intercellular adhesion-induced P40763 activation was mediated by JAK and Src signaling and partially by P00533 signaling . In addition , immunolocalization studies revealed initial formation of phosphorylated P40763 -Y705 at nascent P12830 cell junctions with eventual translocation to the nucleus in cell aggregates . Adhesion-mediated P35610 activation in monolayer and cell aggregate cultures required functional P12830 . These results indicate that , in HNSCC cells , cadherin-mediated intercellular adhesion induces P35610 signaling that may modulate cell survival and resistance to apoptosis during tumor progression . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Preclinical and clinical evaluations of DB01269 , a fully human anti-epidermal growth factor receptor antibody . The epidermal growth factor receptor ( P00533 ) is a transmembrane glycoprotein , with an extracellular ligand-binding domain and intracellular tyrosine kinase domain . Ligand binding induces P00533 dimerization and autophosphorylation on several tyrosine residues in the intracellular domain , leading to mitogenic signal transduction . P00533 overexpression correlates with a poor prognosis and is often associated with malignant transformation in a variety of epithelial cancers . DB01269 is a high-affinity ( dissociation constant K(D) = 5 x 10(-11) M ) fully human IgG2 monoclonal antibody against human P00533 . DB01269 binds P00533 and blocks receptor binding of P01133 and transforming growth factor-alpha , inhibiting P00533 tyrosine phosphorylation and tumor cell activation . DB01269 prevents tumor formation and eradicates large , established A431 tumors in xenograft models . Tumor growth inhibition occurs at relatively low doses , without concomitant chemotherapy or radiotherapy . When combined with chemotherapeutic agents , DB01269 has resulted in additive antitumor activity . A Phase I clinical trial has demonstrated activity in several tumor types , and the results from a Phase II trial for renal cell cancer also showed modest activity . Therapy was generally well tolerated without statistically significant adverse events . Monoclonal antibody blockade of P00533 represents a new and exciting direction in cancer therapy . JAK and P35610 proteins are expressed and activated by P01579 in rat pancreatic acinar cells . The development of acute pancreatitis ( AP ) is triggered by acinar events , but the subsequent extra-acinar events , particularly a distinct immune response , appear to determine its severity . Cytokines modulate this immune response and are derived not only from immunocytes but also from pancreatic acinar cells . We studied whether pancreatic acinar cells were also capable of responding to cytokines . The JAK/ P35610 -pathway represents the main effector for many cytokines . Therefore , expression and regulation of JAK and P35610 proteins were investigated in rat pancreatic acinar cells . Western blotting showed expression of P23458 , O60674 , Tyk2 , and P42224 , P52630 , P40763 , P42229 , P42226 . In addition , P42224 was reversibly tyrosine-phosphorylated upon the procedure of acinar cell isolation . In contrast , P40763 -phosphorylation occurred spontaneously after pancreas removal and was not reversible within 8 h . P42224 phosphorylation was also observed upon treatment with P01579 but not upon P01133 , P01375 or P05231 , and inhibited by the O60674 -inhibitor AG-490 . Immunohistochemistry revealed cytoplasmic expression of unphosphorylated P42224 in untreated acinar cells and nuclear translocation of phosphorylated P42224 following P01579 -treatment . Interestingly , although CCK leads to the activation of multiple stress pathways in pancreatic acinar cells , we found no influence of CCK on phosphorylation of P42224 , P40763 , or P42229 in the pancreas . In conclusion , our data provide further evidence that pancreatic acinar cells are able to interact with immune cells . Besides stimulating immune cells via cytokine secretion , acinar cells are in turn capable of responding to P01579 via O60674 and P42224 which may have an impact on the development of AP . Improving disease control in advanced colorectal cancer : DB01269 and cetuximab . Colorectal cancer remains a major public health concern in Europe and North America . It is responsible for one million new cases and half a million deaths per year worldwide . During the past few years new effective treatments have evolved improving the outcome of patients with this disease . Several alternatives are currently available for advanced colorectal cancer patients including different chemotherapeutic regimens ( fluoropyrimidines , irinotecan and oxaliplatin ) and targeted therapies such as bevacizumab and cetuximab . Different combinations achieve a median survival of over 2 years . Intense efforts focus on identifying agents targeting growth factor receptors , signal transduction pathways or angiogenesis mediators . One of the last available drugs for the management of advanced colorectal cancer is panitumumab , a well-tolerated and effective anti- P00533 monoclonal antibody approved as a single agent in chemotherapy refractory patients . We discuss the current evidence supporting panitumumab for metastatic colorectal cancer treatment , potential predictive biomarkers and ongoing clinical trials with different combinations including panitumumab . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . P01308 suppresses IKs ( P51787 / P15382 ) currents , which require β-subunit P15382 . Abnormal QT prolongation in diabetic patients has become a clinical problem because it increases the risk of lethal ventricular arrhythmia . In an animal model of type 1 diabetes mellitus , several ion currents , including the slowly activating delayed rectifier potassium current ( IKs ) , are altered . The IKs channel is composed of P51787 and P15382 subunits , whose genetic mutations are well known to cause long QT syndrome . Although insulin is known to affect many physiological and pathophysiological events in the heart , acute effects of insulin on cardiac ion channels are poorly understood at present . This study was designed to investigate direct electrophysiological effects of insulin on IKs ( P51787 / P15382 ) currents . P51787 and P15382 were co-expressed in Xenopus oocytes , and whole cell currents were measured by a two-microelectrode voltage-clamp method . Acute application of insulin suppressed the P51787 / P15382 currents and phosphorylated Akt and extracellular signal-regulated kinase ( P29323 ) , the two major downstream effectors , in a concentration-dependent manner . Wortmannin ( 10(-6) M ) , a phosphoinositide 3-kinase ( PI3K ) inhibitor , attenuated the suppression of the currents and phosphorylation of Akt by insulin , whereas U0126 ( 10(-5) M ) , a mitogen-activated protein kinase kinase ( MEK ) inhibitor , had no effect on insulin-induced suppression of the currents . In addition , insulin had little effect on P51787 currents without P15382 , which indicated an essential role of P15382 in the acute suppressive effects of insulin . Mutagenesis studies revealed amino acid residues 111-118 within the distal third C-terminus of P15382 as an important region . P01308 has direct electrophysiological effects on IKs currents , which may affect cardiac excitability .

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DB05773 is highly effective in preclinical models of P04626 -positive gastric cancer . BACKGROUND : A novel antibody-drug conjugate ( trastuzumab-DM1 , DB05773 ) is currently in clinical trials for patients with trastuzumab resistant P04626 -positive breast cancer . Since no clinical data is available from gastric cancer , we studied DB05773 on P04626 -positive human gastric cancer cells and xenograft tumors . METHODS : Effects of DB05773 were studied in four P04626 -positive gastric cancer cell lines ( N-87 , OE-19 , SNU-216 and MKN-7 ) in vitro . Xenograft tumors from N-87 and OE-19 were studied to determine the effect of DB05773 in vivo . RESULTS : DB05773 was found more effective than trastuzumab in N-87 and OE-19 , and moderately effective in MKN-7 cells . On SNU-216 cells both trastuzumab and DB05773 showed limited efficacy . In xenograft tumor experiments , complete pathological response was observed in all OE-19 xenografted mice and in half of the N-87 xenografted mice . The results were equally good irrespective of the tumor burden at therapy initiation , or preceding trastuzumab treatment . DB05773 treatment showed direct effects ( apoptotic cell death and aberrant mitosis ) as well as it mediated antibody-dependent cellular cytotoxicity ( ADCC ) . CONCLUSIONS : DB05773 showed a promising anti-tumor effect in P04626 -positive gastric cancer cell lines in vitro and in vivo , even in tumors which had developed resistance to trastuzumab . DB05773 therapy may warrant clinical trials for P04626 -positive gastric cancer patients . DB00072 emtansine in the treatment of P04626 -positive metastatic breast cancer in Japanese patients . Anti- P04626 agents , such as trastuzumab , lapatinib , trastuzumab emtansine ( DB05773 ) , and pertuzumab , are standard agents in the treatment of breast cancer overexpressing the human epidermal growth factor receptor 2 ( P04626 ) . DB00072 is the first approved P04626 -targeted agent . Subsequent developments include agents with different mechanisms . In this paper , we review the results of clinical trials of DB05773 , a new anti- P04626 agent , with a focus on Japanese patients with breast cancer . On the basis of results from a Phase I study ( JO22591 ) , the maximum tolerated dose was determined to be 3.6 mg/kg every 3 weeks for both Japanese and western patients . In a Phase II study ( JO22997 ) , the overall response rate was 38.4 % ( 90 % confidence interval 28.8-48.6 ) . DB05773 was well tolerated in Japanese patients ; however , the incidence of grade 3 or 4 thrombocytopenia was higher than that observed in earlier western studies , but was not associated with clinically important symptoms . Pharmacokinetic parameters for DB05773 and its metabolites were consistent with those reported previously from a Phase I or II study in non-Japanese patients , and the data obtained showed no suggestion of ethnic differences . Several Phase III studies of DB05773 are ongoing throughout the world , including in Japanese patients with breast cancer . The adjuvant treatment of P04626 -positive breast cancer . About 15-20 % of patients with early stage breast cancer present with tumors that have overexpression or amplification of the human epidermal growth factor receptor 2 ( P04626 ) gene . Before 2005 , these individuals had an increased risk of recurrence and death , but since then their outcomes have substantially improved with the adoption in most countries of adjuvant trastuzumab as a standard component of therapy for P04626 -positive early-stage breast cancer . Consequently , access to high-quality and accurate P04626 testing methods is critical to accurately determine P04626 status , guide treatment decisions , and ultimately improve clinical outcomes . In 2012 , the humanized monoclonal anti-HER antibody trastuzumab was the only approved P04626 -targeted therapy in the adjuvant setting . Data from the first generation of trials combining it with various chemotherapy regimens showed significant improvements in disease-free and overall survival ( DFS/OS ) . Based on results from five randomized clinical trials , a trastuzumab-containing regimen for up to 1 year is now considered standard for all patients with P04626 -positive tumors larger than 1 cm in size who would have fulfilled eligibility to those studies , and this recommendation is sometimes extended to patients with stage I tumors greater than 0.5 cm ( T1b ) . Second generation adjuvant studies with other P04626 -targeted agents like lapatinib and pertuzumab are ongoing , and newer drugs like DB05773 and neratinib are being actively tested in the metastatic setting . Influence of mammographic density on the diagnostic accuracy of tumor size assessment and association with breast cancer tumor characteristics . PURPOSE : The accuracy of breast cancer staging involves the estimation of the tumor size for the initial decision-making in the treatment . We investigated the accuracy of tumor size estimation and the association between tumor characteristics and breast density ( BD ) . MATERIALS AND METHODS : A total of 434 women with a primary diagnosis of breast cancer were included in this prospective study at a specialist breast unit . Estimated tumor characteristics included tumor size , nodal status , estrogen/progesterone receptor status , Ki-67 , P04626 /neu , vascular invasion . Radiomorphological data included tumor size as assessed by mammography , breast ultrasonography , and clinical examination , and American College of Radiology ( P10323 ) categories for BD . RESULTS : BD did not have a significant impact on the assessment of tumor size using breast ultrasound ( deviation from P10323 categories 1-4 : 0.55-0.68 cm ; P=0.331 ) . The deviation in mammography was significantly different dependent on BD ( 0.42-0.9 cm ; P < 0.001 ) . The clinical examination was not affected by BD . Age and tumor size were the only parameters associated with a denser breast in the multivariate analysis . Older women were less likely to have dense breasts ( odds ratio 0.157 for women aged > or=70 years ) , and patients with larger tumors were less likely to have dense breasts ( adjusted OR 0.36 for tumors > 2 cm ) . CONCLUSION : Breast ultrasonography is more accurate than mammography for assessing tumor size in breasts with a higher BD . The difference in tumor size assessment needs to be taken into consideration in the design of clinical trials and treatment decisions . The potential for trastuzumab emtansine in human epidermal growth factor receptor 2 positive metastatic breast cancer : latest evidence and ongoing studies . The treatment of breast cancer that is driven by amplification and overexpression of human epidermal growth factor receptor 2 ( P04626 ) has been drastically improved by the development of P04626 -targeted therapies including trastuzumab and lapatinib . While outcomes for patients diagnosed with P04626 -positive breast cancer have been greatly impacted by these therapies , treatment resistance is common and toxicity to standard regimens remains a therapeutic challenge . DB00072 emtansine ( DB05773 ) is a novel antibody drug conjugate that consists of the P04626 -targeted monoclonal antibody , trastuzumab , joined via a stable linker to a derivative of maytansine , a highly potent cytotoxic chemotherapy . While other antibody drug conjugates have been developed clinically , this is the first in its class that maintains the antitumor properties of the P04626 -targeted antibody , trastuzumab , and also avoids release of the chemotherapy until the molecule is taken up inside the P04626 -overexpressing cancer cell . Several phase I studies have shown DB05773 is safe , tolerable and has activity in trastuzumab- and lapatinib-pretreated breast cancer . Moreover , phase II studies are now being reported that confirm its safety and clinical efficacy in both the frontline and heavily pretreated settings . Preliminary data from phase II studies evaluating its use in combination with other cytotoxics have also been reported and several large phase III trials are underway to evaluate its use in the P04626 -positive metastatic breast cancer setting . This paper aims to provide a detailed review of the preclinical and clinical evidence relating to the mechanism of action , efficacy and safety of DB05773 for the treatment of P04626 -positive breast cancer . DB05773 , a novel antibody-drug conjugate , is highly effective against primary P04626 overexpressing uterine serous carcinoma in vitro and in vivo . Amplification of c-erbB2 has been reported in over 30 % of uterine serous carcinoma ( USC ) and found to confer poor survival because of high proliferation and increased resistance to therapy . In this study , we evaluated for the first time DB00072 emtansine ( DB05773 ) , a novel antibody-drug conjugate , against multiple epidermal growth factor receptor-2 ( P04626 ) -positive USC cells in vitro followed by developing a supportive in vivo model . Fifteen primary USC cell lines were assessed by immunohistochemistry ( IHC ) and flow cytometry for P04626 protein expression . C-erbB2 gene amplification was evaluated using fluorescent in situ hybridization . Sensitivity to DB05773 and trastuzumab ( T ) -induced antibody-dependent cell-mediated cytotoxicity was evaluated in 5-h chromium release assays . DB05773 and T cytostatic and apoptotic activities were evaluated using flow-cytometry-based proliferation assays . In vivo activity of DB05773 versus T in USC xenografts in SCID mice was also evaluated . High levels of P04626 protein overexpression and P04626 gene amplification were detected in 33 % of USC cell lines . DB05773 was considerably more effective than trastuzumab in inhibiting cell proliferation and in causing apoptosis ( P = 0.004 ) of USC showing P04626 overexpression . Importantly , DB05773 was highly active at reducing tumor formation in vivo in USC xenografts overexpressing P04626 ( P = 0.04 ) and mice treated with TDM-1 had significantly longer survival when compared to T-treated mice and control mice ( P ≤ 0.0001 ) . DB05773 shows promising antitumor effect in P04626 -positive USC cell lines and USC xenografts and its activity is significantly higher when compared to T . DB05773 may represent a novel treatment option for P04626 -positive USC patients with disease refractory to trastuzumab and traditional chemotherapy . Systemic treatment of P04626 -positive metastatic breast cancer : a systematic review . AIM : We aimed to systematically review and summarize data from the available clinical trials that examined the treatment of P04626 -positive metastatic breast cancer . METHODS : We reviewed phase 2 and 3 studies in which an anti- P04626 agent was used in one or both arms of the study . While formal meta-analysis was not possible for such a heterogeneous group of trials , resulting forest plots outline some generalizable findings . RESULTS : There is strong evidence that the addition of an anti- P04626 agent to standard chemo- or endocrine therapy improves clinically relevant measurable outcomes . There is also consistent evidence that initial treatment with trastuzumab alone ( and subsequent use of a cytotoxic ) is inferior to the initial combination of trastuzumab plus chemotherapy , and that either DB05773 or dual anti- P04626 agents are superior to single anti- P04626 agent regimens . There is no strong evidence that the use of more than one cytotoxic agent together with an anti- P04626 agent confers any benefit over a single cytotoxic , anti- P04626 combination . CONCLUSION : This review provides a strong evidence base for current clinical practice with a discussion of treatment in the Australian setting . Triggering P25942 on endothelial cells contributes to tumor growth . Inflammatory cells can either promote or inhibit tumor growth . Here we studied whether P25942 , a key molecule for adaptive immune response , has any role in mammary carcinogenesis of BALB/NeuT transgenic tumor-prone mice . We transferred the P04626 /neu oncogene into P25942 -null background to obtain the P25942 -KO/NeuT strain . P25942 -KO/NeuT mice showed delayed tumor onset and reduced tumor multiplicity . BM ( BM ) transplantation experiments excluded a role of BM-derived cells in the reduced tumorigenicity associated with P25942 deficiency . Rather , P25942 expressed by endothelial cells ( ECs ) takes part to the angiogenic process . Accordingly , large vessels , well organized around the tumor lobular structures , characterize BALB/NeuT tumors , whereas tiny numerous vessels with scarce extracellular matrix are dispersed in the parenchyma of poorly organized P25942 -KO/NeuT tumors . Activated platelets , which may interact with and activate ECs , are a possible source of P29965 . Their localization within tumor vessels prompted the idea of treating BALB/NeuT and P25942 -KO/NeuT mice chronically with the anti-platelet drug clopidogrel , known to inhibit platelet P29965 expression . Treatment of BALB/NeuT mice reduced tumor growth to a level similar to P25942 -deficient mice , whereas P25942 -KO/NeuT mice treated or not showed the same attenuated tumor outgrowth , indicating that activated platelets are the likely source of P29965 in this model . Collectively , these data point to a participation of P25942 / P29965 in the angiogenic processes associated with mammary carcinogenesis of BALB/NeuT mice . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Safety and efficacy of the combination of DB05773 with radiotherapy of the central nervous system in a patient with P04626 -positive metastatic breast cancer : case study and review of the literature . Approximately 35 % of patients with confirmed P04626 breast cancer progress to metastases of the central nervous system ( CNS ) . Total cerebral radiotherapy is considered as standard treatment for these cases ; however , studies have shown that some chemotherapy drugs can be used during radiotherapy without significantly increasing its toxicity . In this article , we report the case of a patient with P04626 -positive breast cancer who showed isolated progression of the illness in the CNS , which was observed during the treatment period using DB05773 concomitantly with radiotherapy of the CNS without apparent toxicity of the combination and keeping the illness controlled . Through a review of the literature on the use of radiotherapy and chemotherapy with DB05773 for the treatment of cerebral metastases in P04626 -positive breast cancer , we describe the efficacy and tolerance of the concomitant application of these treatments . Multifocal synchronous mucinous adenocarcinomas arising in congenital pulmonary airway malformation : a case report with molecular study . AIMS : Congenital pulmonary airway malformation ( CPAM ) is a rare developmental anomaly of the lung . Here , we report a case of mucinous adenocarcinoma arising in CPAM . A 23-month-old boy underwent a thoracoscopic lobectomy of the left upper lobe of the lung based on a presumptive diagnosis of asymptomatic CPAM , found in antenatal sonogram . METHODS AND RESULTS : Histologically , the lesion was consistent with CPAM , Stocker type I . In addition , multiple foci ranging from mucinous epithelial hyperplasia to mucinous adenocarcinoma were detected . All lesions shared the same immunoprofile with the expression of cytokeratin ( CK ) 20 , P98088 , and human epidermal growth factor receptor2 ( P04626 ) , but were negative for CK7 , transcription factor 1 ( Q15669 -1 ) , P15941 , Q99626 , P15056 ( VE1 ) and anaplastic lymphoma kinase ( Q9UM73 ) . K- DB01367 point mutation ( G12V ) was also detected in all micro-dissected mucinous lesions but P00533 mutation was not found . All lesions were consistent with mucinous adenocarcinoma . The patient 's clinical course has been uneventful during the 12-months follow-up period . CONCLUSIONS : This interesting case demonstrated that multiple foci in CPAM can synchronously transform into malignancies . Dual targeting of P04626 -positive cancer with trastuzumab emtansine and pertuzumab : critical role for neuregulin blockade in antitumor response to combination therapy . PURPOSE : Targeting P04626 with multiple P04626 -directed therapies represents a promising area of treatment for P04626 -positive cancers . We investigated combining the P04626 -directed antibody-drug conjugate trastuzumab emtansine ( DB05773 ) with the P04626 dimerization inhibitor pertuzumab ( Perjeta ) . EXPERIMENTAL DESIGN : Drug combination studies with DB05773 and pertuzumab were performed on cultured tumor cells and in mouse xenograft models of P04626 -amplified cancer . In patients with P04626 -positive locally advanced or metastatic breast cancer ( mBC ) , DB05773 was dose-escalated with a fixed standard pertuzumab dose in a 3+3 phase Ib/II study design . RESULTS : Treatment of P04626 -overexpressing tumor cells in vitro with DB05773 plus pertuzumab resulted in synergistic inhibition of cell proliferation and induction of apoptotic cell death . The presence of the P21860 ligand , heregulin ( Q99988 β ) , reduced the cytotoxic activity of DB05773 in a subset of breast cancer lines ; this effect was reversed by the addition of pertuzumab . Results from mouse xenograft models showed enhanced antitumor efficacy with DB05773 and pertuzumab resulting from the unique antitumor activities of each agent . In patients with mBC previously treated with trastuzumab , lapatinib , and chemotherapy , DB05773 could be dosed at the maximum tolerated dose ( MTD ; 3.6 mg/kg every 3 weeks ) with standard dose pertuzumab . Adverse events were mostly grade 1 and 2 , with indications of clinical activity . CONCLUSIONS : Dual targeting of P04626 with the combination of DB05773 and pertuzumab in cell culture and mouse xenograft models resulted in enhanced antitumor activity . In patients , this combination showed an encouraging safety and tolerability profile with preliminary evidence of efficacy . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Enhanced antitumor activity of trastuzumab emtansine ( DB05773 ) in combination with pertuzumab in a P04626 -positive gastric cancer model . Human epidermal growth factor receptor 2 ( P04626 ) -targeted therapy by trastuzumab has become increasingly important for treating P04626 -positive cancers , and trastuzumab emtansine ( DB05773 ) is expected to serve as an effective alternative to trastuzumab . DB06366 , a P04626 dimerization inhibitor , showed prolonged progression-free survival when used with trastuzumab for P04626 -positive breast cancer . In this study , we investigated the effect of combining DB05773 and pertuzumab on xenografted gastric tumors . DB05773 as a single agent showed significant antitumor activity in all the three P04626 -high expression tumor models tested ( NCI-N87 , P35240 and 4-1ST ) but was ineffective against two P04626 -low expression tumors ( SNU-16 and MKN-28 ) . Using the DB05773 -sensitive NCI-N87 model , the combination efficacy of DB05773 and pertuzumab was elucidated . The combination induced significant tumor regression , whereas DB05773 or pertuzumab alone did not . In cultured NCI-N87 cells stimulated with epidermal growth factor ( P01133 ) or heregulin-α , concomitant treatment of DB05773 and pertuzumab significantly inhibited proliferation and increased caspase 3/7 activity compared to either agent alone . Only the combination significantly inhibited the phosphorylation of P00533 or P21860 , and its downstream factor AKT . Suppressed P21860 phosphorylation by the combination was also seen in the NCI-N87 xenografted tumors . Compared to single agent treatments , the combination treatment significantly enhanced antibody-dependent cellular cytotoxicity ( ADCC ) against NCI-N87 cells . These findings suggest that DB05773 in combination with pertuzumab shows significant antitumor activity by increasing AKT signal inhibition and ADCC in P04626 -positive gastric cancers . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Why your preferred targeted drugs may become unaffordable . DB00072 , a monoclonal antibody directed at the P04626 receptor , is one of the most impressive targeted drugs developed in the last two decades . Indeed , when given in conjunction with chemotherapy , it improves the survival of women with P04626 positive breast cancer , both in advanced and in early disease . Its optimal duration , however , is poorly defined in both settings with a significant economic impact in the adjuvant setting where the drug is arbitrarily given for 1 year . This article reviews current attempts at shortening this treatment duration , emphasizing the likelihood of inconclusive results and , therefore , the need to investigate this important variable as part of the initial pivotal trials and with the support of public health systems . Failure to do so has major consequences on treatment affordability . Ongoing adjuvant trials of dual P04626 blockade , using trastuzumab in combination with a second anti- P04626 agent , and trials of the antibody-drug conjugate DB05773 ( trastuzumab-emtansine ) have to all be designed with 12 months of targeted therapy . Nursing perspectives on trastuzumab emtansine for the treatment of metastatic breast cancer . Increased understanding of the molecular composition of breast cancer tumors has led to the development of targeted anticancer agents . Novel therapies directed against human epidermal growth factor receptor 2 ( P04626 ) in breast cancer have been developed . One such agent , trastuzumab emtansine ( DB05773 ) , is an antibody drug conjugate that has been shown to be effective in the treatment of women with P04626 -positive breast cancer . Phase I and II studies have determined a maximum tolerated dose , and several phase Ib/II , II , and III studies have shown improved tolerability and efficacy compared with the combination of trastuzumab and chemotherapy . The most concerning grade 3 or higher adverse events associated with DB05773 include thrombocytopenia and transaminitis . To ensure that these adverse events do not delay or interrupt treatment , oncology nurses need to familiarize themselves with these risks and their management . This article reviews the clinical development of DB05773 and its usage , with a focus on the nurse 's role in preventing and managing adverse events associated with DB05773 therapy . Population pharmacokinetics of trastuzumab emtansine ( DB05773 ) , a P04626 -targeted antibody-drug conjugate , in patients with P04626 -positive metastatic breast cancer : clinical implications of the effect of covariates . PURPOSE : DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate comprising the humanized monoclonal antibody trastuzumab linked to DM1 , a highly potent cytotoxic agent . A population pharmacokinetic ( PK ) analysis was performed to estimate typical values and interindividual variability of DB05773 PK parameters and the effects of clinically relevant covariates . METHODS : Serum samples were collected from 671 patients with human epidermal growth factor receptor 2-positive locally advanced or metastatic breast cancer ( MBC ) who received single-agent DB05773 in five phase I to phase III studies . Nonlinear mixed-effects modeling with the first-order conditional estimation method was used . RESULTS : A linear two-compartment model with first-order elimination from the central compartment described DB05773 PKs in the clinical dose range . DB05773 elimination clearance was 0.676 L/day , volume of distribution in the central compartment ( V c ) was 3.127 L , and terminal elimination half-life was 3.94 days . Age , race , region , and renal function did not influence DB05773 PK . Given the low-to-moderate effect of all statistically significant covariates on DB05773 exposure , none of these covariates is expected to result in a clinically meaningful change in DB05773 exposure . CONCLUSIONS : DB05773 PK properties are consistent and predictable in patients . A further refinement of dose based on baseline covariates other than body weight for the current 3.6 mg/kg regimen would not yield clinically meaningful reductions in interindividual PK variability in patients with MBC . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Live cell off-target identification of lapatinib using ligand-directed tosyl chemistry . We demonstrate that ligand-directed tosyl ( DB01265 ) chemistry is applicable to off-target identification in live cells . DB01259 ( Lap ) -based DB01265 reagents not only labeled a receptor tyrosine kinase , P04626 , target protein , but also the protein disulfide isomerase ( P07237 ) that should be an off-target protein for Lap . DB05773 ( DB05773 ) in human epidermal growth factor receptor 2 ( P04626 ) -positive metastatic breast cancer : latest evidence and clinical potential . In February 2013 , ado-trastuzumab emtansine ( DB05773 , Kadcyla® ) received regulatory approval in the United States for treatment-refractory human epidermal growth factor receptor 2 ( P04626 ) positive metastatic or locally advanced breast cancer based on results from EMILIA , a large phase III trial that compared standard of care lapatinib plus capecitabine to DB05773 . Several other studies have been reported in the metastatic setting and multiple trials are ongoing or planned in the neoadjuvant , adjuvant and advanced disease settings . Here we provide an updated and comprehensive review of clinical trials evaluating DB05773 , discuss management of toxicity associated with this drug , propose potential mechanisms of resistance and offer practical considerations for the treating oncologist . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Phase I and pharmacokinetic study of trastuzumab emtansine in Japanese patients with P04626 -positive metastatic breast cancer . OBJECTIVE : DB00072 emtansine ( DB05773 ) , an antibody-drug conjugate composed of the cytotoxic agent DM1 conjugated to trastuzumab via a stable thioether linker , has shown clinical activity in human epidermal growth factor receptor 2-positive metastatic breast cancer patients . This study evaluated the maximum tolerated dose , toxicity and pharmacokinetics of trastuzumab emtansine in Japanese breast cancer patients . METHODS : Inoperable advanced or recurrent human epidermal growth factor receptor 2-positive breast cancer patients were administered trastuzumab emtansine intravenously at a dose of 1.8 , 2.4 or 3.6 mg/kg every 3 weeks . The maximum tolerated dose was estimated using the continual reassessment method . RESULTS : This study enrolled 10 patients who were administered trastuzumab emtansine for a median of seven cycles . The dose-limiting toxicity was Grade 3 elevation of aspartate aminotransferase/alanine aminotransferase at the 2.4 mg/kg dose level . The maximum tolerated dose was estimated to be 3.6 mg/kg because at the point when dose-limiting toxicity was evaluable in 10 patients , the probability of dose-limiting toxicity estimated using the continual reassessment method was closest to 25 % at a dose of 3.6 mg/kg and this was unchanged by the results for patients enrolled after that . The most frequent adverse events were nausea , arthralgia , fever , fatigue and decreased appetite . Adverse events were generally tolerable . The maximum concentration and area under the concentration-time curve increased linearly with the dose . CONCLUSIONS : DB00072 emtansine up to 3.6 mg/kg was well tolerated by Japanese breast cancer patients . Although thrombocytopenia and hepatotoxicity tended to be more severe than was seen in Western patients in previous trastuzumab emtansine trials , those adverse events recovered without special supportive treatment . DB05773 : a P04626 -positive targeted antibody-drug conjugate . OBJECTIVE : To review the pharmacology , pharmacokinetics , efficacy , adverse effects , drug-drug interactions , dosage and administration , and formulary considerations for ado-trastuzumab emtansine . DATA SOURCES : Sources of information were identified through a PubMed search ( 1966 to June 2014 ) using the key terms ado-trastuzumab emtansine , trastuzumab-DM1 , trastuzumab-MCC-DM1 , and DB05773 . Other information was obtained from clinicaltrials.gov , product labeling , and press releases . STUDY SELECTION AND DATA EXTRACTION : All English-language clinical trials and abstracts evaluating ado-trastuzumab emtansine in humans were reviewed for inclusion . DATA SYNTHESIS : Overexpression or amplification of human epidermal growth factor receptor 2 ( P04626 ) occurs in approximately 20 % of breast cancers and is associated with more aggressive tumors and poorer prognosis in the absence of treatment . Although effective therapies for the initial management of P04626 -positive metastatic breast cancer ( MBC ) exist , many patients will experience disease progression . Most second-line therapies are associated with either significant toxicities or limited improvements in overall survival ( OS ) . DB05773 is a P04626 -positive directed antibody drug conjugate ( ADC ) approved in February 2013 . In phase III clinical trials comparing the efficacy and safety of ado-trastuzumab emtansine with lapatinib-capecitabine or physician 's choice , ado-trastuzumab emtansine had a better tolerability profile and improved progression-free survival compared with lapatinib-capecitabine or physician 's choice and increased OS compared with lapatinib-capecitabine . CONCLUSION : DB05773 is a novel ADC effective for P04626 -positive MBC in patients previously treated with trastuzumab , lapatinib , and a taxane . Further studies will determine its use in the adjuvant and neoadjuvant setting and in combination with pertuzumab . Is the Improved Efficacy of DB00072 and DB01259 Combination Worth the Added Toxicity ? A Discussion of Current Evidence , Recommendations , and Ethical Issues Regarding Dual P04626 -Targeted Therapy . Following FDA approval of trastuzumab in 1998 and lapatinib in 2007 , several clinical studies have addressed the question of whether trastuzumab and lapatinib combination therapy is better than trastuzumab alone in the metastatic breast cancer and neoadjuvant setting . In this review , updated to September 2012 , we focus on the relevant clinical trials that address this question and , based on the available data , reach conclusions regarding a rational and reasonably individualized approach to the management of P04626 + breast cancer . With the FDA approval of pertuzumab in June 2012 and the likely approval of DB05773 approaching , several ethical issues overshadow the excitement oncologists have for these new treatment options . We discuss the potential evolution of highly active anti- P04626 therapy ( HAAHT ) as an optimal treatment paradigm for P04626 + breast cancer . Additionally , we review lessons learned from the evolution of HAART for HIV treatment . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Cellular mechanisms of the hemostatic effects of desmopressin ( DB00035 ) . The synthetic analog of vasopressin desmopressin ( DB00035 ) is widely used for the treatment of patients with von Willebrand disease ( VWD ) , hemophilia A , several platelet disorders , and uremic bleeding . DB00035 induces an increase in plasma levels of P04275 ( P04275 ) , coagulation factor VIII ( FVIII ) , and tissue plasminogen activator ( t-PA ) . It also has a vasodilatory action . In spite of its extensive clinical use , its cellular mechanism of action remains incompletely understood . Its effect on P04275 and t-PA as well as its vasodilatory effect are likely explained by a direct action on the endothelium , via activation of endothelial vasopressin P30518 receptor and DB02527 -mediated signaling . This leads to exocytosis from Weibel Palade bodies where both P04275 and t-PA are stored , as well as to nitric oxide ( NO ) production via activation of endothelial NO synthase . The mechanism of action of DB00035 on FVIII plasma levels remains to be elucidated . The hemostatic effect of DB00035 likely involves additional cellular effects that remain to be discovered . [ DB05773 and pertuzumab : emerging anti- P04626 therapeutics ] . P04626 -targeted therapy for P04626 -positive breast cancer is one of the success stories in medical oncology . DB00072 , a humanized monoclonal antibody , was the first approved P04626 -targeted agent . Subsequent developments include agents with different mechanisms , such as lapatinib , a tyrosine kinase inhibitor . We describe here the results of late-phase clinical trials of two newly-available anti- P04626 agents , DB05773 and pertuzumab . Relation between hemostatic parameters and prognostic/predictive factors in breast cancer . BACKGROUND : In our study , we searched for a relation between various prognostic and predictive factors and hemostatic parameters . METHODS : One hundred women with newly diagnosed breast cancer after surgery were included . Patients did not receive systemic therapy or radiotherapy . The control group included 100 healthy , age-matched women . In the patient group , age , menopausal status , tumor size , grade , axillary lymph node status , steroid receptor status , p53 , and P04626 /neu were evaluated . Plasma levels of factor VIII , factor IX , D-dimer , fibrinogen , protein C , protein S , P04275 , and antithrombin III were measured in both groups . RESULTS : Plasma levels of factor VIII , factor IX , P04275 , and CRP in patients with breast cancer were higher than those in controls . Protein S levels in patients were lower than in controls . There was no significant difference in other hemostatic parameters between the groups . In patients with axillary lymph node metastasis , factor VIII levels were significantly higher than in node-negative patients . There was a strong correlation between axillary lymph node status , number of metastatic nodes , and factor VIII levels . There was no correlation between factor VIII levels and CRP . Factor VIII levels were higher in the group having high P04626 /neu ( 3+ ) than in the group with negativity for P04626 /neu . CONCLUSION : There was a strong correlation between axillary lymph node involvement , number of metastatic nodules , overexpression of P04626 /neu , hemostatic parameters , and factor VIII levels . Our study showed that factor VIII level measurement can provide additional data for evaluation of breast cancer patients ' prognosis . Emerging therapeutic targets in bladder cancer . Treatment of muscle invasive urothelial bladder carcinoma ( BCa ) remains a major challenge . Comprehensive genomic profiling of tumors and identification of driver mutations may reveal new therapeutic targets . This manuscript discusses relevant molecular drivers of the malignant phenotype and agents with therapeutic potential in BCa . Small molecule pan-FGFR inhibitors have shown encouraging efficacy and safety results especially among patients with activating FGFR mutations or translocations . P42345 inhibitors for patients with Q92574 mutations and concomitant targeting of PI3K and MEK represent strategies to block PI3K/AKT/ P42345 pathway . Encouraging preclinical results with ado-trastuzumab emtansine ( DB05773 ) exemplifies a new potential treatment for P04626 -positive BCa along with innovative bispecific antibodies . Inhibitors of cell cycle regulators ( aurora kinase , polo-like kinase 1 , and cyclin-dependent kinase 4 ) are being investigated in combination with chemotherapy . Early results of clinical studies with anti- P16410 and anti- Q9NZQ7 are propelling immune modulating drugs to the forefront of emerging treatments for BCa . Collectively , these novel therapeutic targets and treatment strategies hold promise to improve the outcome of patients afflicted with this malignancy . Breast cancer brain metastases responding to lapatinib plus capecitabine as second-line primary systemic therapy . Brain metastases ( BM ) are diagnosed in up to 40 % of P04626 -positive breast cancer patients . Standard treatment includes local approaches such as whole-brain radiotherapy ( WBRT ) , radiosurgery , and neurosurgery . The landscape trial established primary systemic therapy as an effective and safe alternative to WBRT in selected patients with Her2-positive BM . We aim to further focus on the role of systemic therapy in oligosymptomatic patients by presenting this case report . We report on a 50-year-old patient diagnosed with multiple BM 5 years after early breast cancer diagnosis . As the patient was asymptomatic and had a favorable diagnosis-specific P02724 score , she received primary systemic treatment with DB05773 . She achieved partial remission within the brain for eight treatment cycles and then progressed despite stable extracranial disease . As the patient remained asymptomatic and refused WBRT , we decided upon trastuzumab , lapatinib plus capecitabine as second-line therapy . Another partial remission of BM was observed ; to date , she has received 11 treatment cycles without any sign of disease progression . In this case , WBRT was delayed by at least 14 months , again indicating the activity of systemic treatment in BM . Apparently , in selected patients , BM can be controlled with multiple lines of systemic therapy similar to extracranial disease . Further investigation of systemic treatment approaches is therefore warranted . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . The food colorant erythrosine is a promiscuous protein-protein interaction inhibitor . Following our observation that erythrosine B ( FD & C Red No. 3 ) is a relatively potent inhibitor of the P01375 -R-TNFα and P25942 -CD154 protein-protein interactions , we investigated whether this inhibitory activity extends to any other protein-protein interactions ( PPI ) as well as whether any other approved food colors possess such inhibitory activity . We found erythrosine , a poly-iodinated xanthene dye , to be a non-specific promiscuous inhibitor of a number of PPIs within the tumor necrosis factor superfamily ( P01375 -R-TNFα , P25942 -CD154 , Q96RJ3 - Q9Y275 , Q9Y6Q6 - O14788 , OX40- P23510 , 4-1BB- P41273 ) as well as outside of it ( P01133 -R- P01133 ) with a remarkably consistent median inhibitory concentration ( IC(50) ) in the 2-20 μM ( approximately 2-20mg/L ) range . In agreement with this , erythrosine also showed cellular effects including clear cytotoxic effects around this concentration range ( IC₅₀≈50 μM ) . Among the seven FDA-approved food colorants , only erythrosine showed consistent PPI inhibitory activity in the sub-100 μM range , which might also explain ( at least partially ) why it also has the lowest approved acceptable daily intake ( ADI ) ( 0.1 mg/kg body weight/day ) . Among a number of xanthene structural analogs of erythrosine tested for activity , rose Bengal , a food colorant approved in Japan , showed similar , maybe even more pronounced , promiscuous inhibitory activity , whereas fluorescein was inactive and gallein , phloxine , and eosin were somewhat active in some of the assays . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . Updates on the treatment of human epidermal growth factor receptor type 2-positive breast cancer . PURPOSE OF REVIEW : To review the most recent developments in the treatment of human epidermal growth factor receptor type 2 ( P04626 ) -positive breast cancer with novel P04626 -targeting agents and combinations that have significantly improved clinical outcomes . RECENT FINDINGS : Since the approval of trastuzumab 15 years ago , the natural history of P04626 -positive breast cancer has been altered with improvements in survival for both early and advanced disease with the addition of this agent to standard chemotherapy . The P04626 receptor pathway drives breast cancer growth and aggressiveness , and P04626 -targeted agents can improve survival in early and advanced disease . In the advanced setting , two new drugs have been approved since 2012 , pertuzumab and ado-trastuzumab emtansine ( DB05773 ) , both of which improve survival without any reciprocal increase in toxicity . However , resistance almost always ensues , pointing to the need to understand the driving mechanisms and to biomarkers that will help individualize therapy and point to newer signal transduction and other modulators . SUMMARY : P04626 -positive breast cancer represents a distinct subtype with more aggressive clinical characteristics . P04626 -targeted therapies , usually in combination with chemotherapy , are the standard of care , improving the cure rate in early-stage breast cancer and lengthening survival in the advanced setting . Targeting the P04626 receptor in metastatic breast cancer . The advent of targeted therapies has revolutionized the treatment of certain types of cancer . Identification of molecular targets on cancer cells has led to the design of novel drugs , which either used as single agents or in combination with chemotherapy , has prolonged survival in metastatic disease , or contributed to curative treatment in the adjuvant setting . A literature review was conducted to identify and present current knowledge on the molecular function of the P04626 receptor , its role in the pathogenesis of breast cancer and anti- P04626 targeted drugs in use or under development . Many molecular targets have been identified in breast cancer , with the HER family of receptors being the ones most extensively studied . DB00072 and lapatinib target the P04626 receptor and are approved drugs for the treatment of metastatic breast cancer . Several other targeted agents , including DB05773 , pertuzumab , neratinib , afatinib and ertumaxomab , are currently being tested in vivo as well as in clinical studies . The use of targeted therapies in metastatic breast cancer has improved prognosis , increased survival and dramatically changed the way we treat breast cancer patients today . DB11320 H3 receptors aggravate cerebral ischaemic injury by histamine-independent mechanisms . The role of the histamine H3 receptor ( Q9Y5N1 ) in cerebral ischaemia/reperfusion ( I/R ) injury remains unknown . Here we show that Q9Y5N1 expression is upregulated after I/R in two mouse models . Q9Y5N1 antagonists and Q9Y5N1 knockout attenuate I/R injury , which is reversed by an Q9Y5N1 -selective agonist . Interestingly , P35367 and P25021 antagonists , a histidine decarboxylase ( HDC ) inhibitor and HDC knockout all fail to compromise the protection by Q9Y5N1 blockade . Q9Y5N1 blockade inhibits P42345 phosphorylation and reinforces autophagy . The neuroprotection by Q9Y5N1 antagonism is reversed by 3-methyladenine and siRNA for Atg7 , and is diminished in Atg5⁻/⁻ mouse embryonic fibroblasts . Furthermore , the peptide Tat- Q9Y5N1 (CT414-436) , which blocks Q9Y696 binding with H3Rs , or siRNA for Q9Y696 , further increases I/R-induced autophagy and protects against I/R injury . Therefore , Q9Y5N1 promotes I/R injury while its antagonism protects against ischaemic injury via histamine-independent mechanisms that involve suppressing Q9Y5N1 / Q9Y696 binding-activated autophagy , suggesting that Q9Y5N1 inhibition is a therapeutic target for cerebral ischaemia . A common binding site on the microsomal triglyceride transfer protein for apolipoprotein B and protein disulfide isomerase . The assembly of triglyceride-rich lipoproteins requires the formation in the endoplasmic reticulum of a complex between apolipoprotein B ( apoB ) , a microsomal triglyceride transfer protein ( P55157 ) , and protein disulfide isomerase ( P07237 ) . In the P55157 complex , the amino-terminal region of P55157 ( residues 22-303 ) interacts with the amino-terminal region of apoB ( residues 1-264 ) . Here , we report the identification and characterization of a site on apoB between residues 512 and 721 , which interacts with residues 517-603 of P55157 . P07237 binds in close proximity to this apoB binding site on P55157 . The proximity of these binding sites on P55157 for P07237 and amino acids 512-721 of apoB was evident from studies carried out in a yeast two-hybrid system and by co-immunoprecipitation . The expression of P07237 with P55157 and apoB16 ( residues 1-721 ) in the baculovirus expression system reduced the amount of P55157 co-immunoprecipitated with apoB by 73 % . The interaction of residues 512-721 of apoB with P55157 facilitates lipoprotein production . Mutations of apoB that markedly reduced this interaction also reduced the level of apoB-containing lipoprotein secretion . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Preclinical safety profile of trastuzumab emtansine ( DB05773 ) : mechanism of action of its cytotoxic component retained with improved tolerability . DB00072 emtansine ( DB05773 ) is the first antibody-drug conjugate ( ADC ) approved for patients with human epidermal growth factor receptor 2 ( P04626 ) -positive metastatic breast cancer . The therapeutic premise of ADCs is based on the hypothesis that targeted delivery of potent cytotoxic drugs to tumors will provide better tolerability and efficacy compared with non-targeted delivery , where poor tolerability can limit efficacious doses . Here , we present results from preclinical studies characterizing the toxicity profile of DB05773 , including limited assessment of unconjugated DM1 . DB05773 binds primate ErbB2 and human P04626 but not the rodent homolog c-neu . Therefore , antigen-dependent and non-antigen-dependent toxicity was evaluated in monkeys and rats , respectively , in both single- and repeat-dose studies ; toxicity of DM1 was assessed in rats only . DB05773 was well tolerated at doses up to 40 mg/kg ( ~4400 μg DM1/m(2) ) and 30 mg/kg ( ~ 6000 μg DM1/m(2) ) in rats and monkeys , respectively . In contrast , DM1 was only tolerated up to 0.2mg/kg ( 1600 μg DM1/m(2) ) . This suggests that at least two-fold higher doses of the cytotoxic agent are tolerated in DB05773 , supporting the premise of ADCs to improve the therapeutic index . In addition , DB05773 and DM1 safety profiles were similar and consistent with the mechanism of action of DM1 ( i.e. , microtubule disruption ) . Findings included hepatic , bone marrow/hematologic ( primarily platelet ) , lymphoid organ , and neuronal toxicities , and increased numbers of cells of epithelial and phagocytic origin in metaphase arrest . These adverse effects did not worsen with chronic dosing in monkeys and are consistent with those reported in DB05773 -treated patients to date . Potential mechanisms for thrombocytopenia development with trastuzumab emtansine ( DB05773 ) . PURPOSE : DB00072 -emtansine ( DB05773 ) is an antibody-drug conjugate ( ADC ) comprising the cytotoxic agent DM1 conjugated to trastuzumab with a stable linker . Thrombocytopenia was the dose-limiting toxicity in the phase I study , and grade ≥3 thrombocytopenia occurred in up to 13 % of patients receiving DB05773 in phase III studies . We investigated the mechanism of DB05773 -induced thrombocytopenia . EXPERIMENTAL DESIGN : The effect of DB05773 on platelet function was measured by aggregometry , and by flow cytometry to detect the markers of activation . The effect of DB05773 on differentiation and maturation of megakaryocytes ( MK ) from human hematopoietic stem cells was assessed by flow cytometry and microscopy . Binding , uptake , and catabolism of DB05773 in MKs , were assessed by various techniques including fluorescence microscopy , scintigraphy to detect T-[H(3)]-DM1 and (125)I- DB05773 , and mass spectrometry . The role of FcγRIIa was assessed using blocking antibodies and mutant constructs of trastuzumab that do not bind FcγR . RESULTS : DB05773 had no direct effect on platelet activation and aggregation , but it did markedly inhibit MK differentiation via a cytotoxic effect . Inhibition occurred with DM1-containing ADCs but not with trastuzumab demonstrating a role for DM1 . MKs internalized these ADCs in a P04626 -independent , FcγRIIa-dependent manner , resulting in intracellular release of DM1 . Binding and internalization of DB05773 diminished as MKs matured ; however , prolonged exposure of mature MKs to DB05773 resulted in a disrupted cytoskeletal structure . CONCLUSIONS : These data support the hypothesis that DB05773 -induced thrombocytopenia is mediated in large part by DM1-induced impairment of MK differentiation , with a less pronounced effect on mature MKs . Hypoxic/normoxic preconditioning increases endothelial differentiation potential of human bone marrow CD133+ cells . CD133+ cells are hemangioblasts that have capacity to generate into both hematopoietic and endothelial cells ( ECs ) . Hypoxia/normoxia has shown to be the regulator of the balance between stemness and differentiation . In this study we performed Agilent 's whole human genome oligo microarray analysis and examined the differentiation potential of the bone-marrow-derived CD133+ cells after hypoxic/normoxic preconditioning of CD133+ cells . Results showed that there was no significant increase in erythroid colony forming unit ( CFU-E ) and CFU-granulocyte , erythrocyte , monocyte , and megakaryocyte formation with cells treated under hypoxia/normoxia . However , a significant increment of EC forming unit at 24 h ( 143.2 +/- 8.0 % ) compared to 0 h ( 100 +/- 11.4 % ) was observed in CFU-EC analysis . Reverse transcription-polymerase chain reaction and immunostaining analysis showed that the differentiated cells diminished hematopoietic stem cell surface markers and acquired the gene markers and functional phenotype of ECs . The transcriptome profile revealed a cluster of 232 downregulated and 498 upregulated genes in cells treated for 24 h under hypoxia . The upregulated genes include angiogenic genes , angiogenic growth factor genes , angiogenic cytokine and chemokine genes , as well as angiogenic-positive regulatory genes , including Q14512 , PDGFB , Q16663 , P48061 , P80162 , P05231 , P21246 , O14944 , P04626 , O95136 , P11487 , Q92913 , Q99988 , P05412 , L1CAM , Q02297 , P08138 , and PDGFB . On the other hand , angiogenesis inhibitors and related genes , including P29459 , P98177 , Q9NY15 , and P16035 , are downregulated . Taken together , hypoxic/normoxic preconditioning may lead to the differentiation of CD133+ cells toward endothelial lineage , which may improve the current clinical trial studies . DB05773 for the treatment of human epidermal growth factor receptor 2-positive metastatic breast cancer . PURPOSE : An update on completed and ongoing clinical trials of ado-trastuzumab emtansine for the treatment of metastatic breast cancer ( MBC ) is presented . SUMMARY : DB05773 ( Kadcyla , Genentech ) , the first U.S.-approved antibody-drug conjugate for MBC , is indicated for use as a single-agent therapy in patients with human epidermal growth factor receptor 2 ( P04626 ) -positive MBC who have received prior treatment with unconjugated trastuzumab and a taxane-based regimen . The standard dosage of ado-trastuzumab is 3.6 mg/kg i.v. every three weeks . In completed Phase II or III clinical trials , ado-trastuzumab was found to confer significant survival and quality-of-life benefits . The largest of those trials ( the EMILIA study , n = 991 ) showed that ado-trastuzumab was superior to a regimen of lapatinib plus capecitabine in terms of progression-free survival ( 9.6 months versus 6.4 months , p < 0.001 ) and overall survival ( 30.9 months versus 25.1 months , p < 0.001 ) ; it also had a more favorable tolerability profile , with lower rates of treatment-limiting adverse effects . The most common adverse effects of ado-trastuzumab are thrombocytopenia ( reported in about 12 % of clinical trial participants overall ) and increased transaminase levels . Two ongoing Phase III trials-the TH3RESA study ( slated for completion in June 2015 ) and the MARIANNE study ( estimated completion in 2016 ) -may help determine the optimal role of ado-trastuzumab relative to other P04626 -targeted agents and its potential use as a front-line therapy for both heavily pretreated and treatment-naive patients with MBC . CONCLUSION : With a novel targeted mechanism of action , ado-trastuzumab is an effective treatment option for P04626 -positive MBC in previously treated patient populations . Catabolic fate and pharmacokinetic characterization of trastuzumab emtansine ( DB05773 ) : an emphasis on preclinical and clinical catabolism . DB00072 emtansine ( DB05773 ) is an antibody-drug conjugate in clinical development for the treatment of human epidermal growth factor receptor 2 ( P04626 ) -positive cancers . Herein , we describe a series of studies to assess DB05773 absorption , distribution , metabolism , and excretion ( ADME ) in rats as well as to assess human exposure to DB05773 catabolites . Following administration of unlabeled and radiolabeled DB05773 in female Sprague Dawley rats as a single dose , plasma , urine , bile and feces were assessed for mass balance , profiling and identification of catabolites . In rats , the major circulating species in plasma was DB05773 , while DM1 concentrations were low ( 1.08 to 15.6 ng/mL ) . The major catabolites found circulating in rat plasma were DM1 , [ N-maleimidomethyl ] cyclohexane-1- carboxylate-DM1 ( MCC-DM1 ) , and DB00123 -MCC-DM1 . These catabolites identified in rats were also detected in plasma samples from patients with P04626 -positive metastatic breast cancer who received single-agent DB05773 ( 3.6 mg/kg every 3 weeks ) in a phase 2 clinical study . There was no evidence of tissue accumulation in rats or catabolite accumulation in human plasma following multiple dosing . In rats , DB05773 was distributed nonspecifically to the organs without accumulation . The major pathway of DM1-containing catabolite elimination in rats was the fecal/biliary route , with up to 80 % of radioactivity recovered in the feces and 50 % in the bile . The rat DB05773 ADME profile is likely similar to the human profile , although there may be differences since trastuzumab does not bind the rat P04626 - like receptor . Further research is necessary to more fully understand the DB05773 ADME profile in humans . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . Treating metastatic breast cancer with systemic chemotherapies : current trends and future perspectives . Treatment selection for metastatic breast cancer ( MBC ) is guided by multiple factors , most importantly hormone receptor ( HR ) or P04626 expression , treatment history , and prognostic factors such as short disease-free interval , presence of visceral metastases , performance status , and degree of symptoms . Chemotherapy is indicated as initial therapy for patients with HR-negative disease and following failure of hormonal therapies in HR-positive disease . Patients treated with an anthracycline or a taxane in early-stage settings may no longer be candidates for those drugs in MBC , thus underscoring the need for alternative options . Sequential single-agent therapy or combination therapy are viable strategies . Trials have shown that ixabepilone plus capecitabine significantly improves progression-free survival compared with capecitabine alone in anthracycline- or taxane-pretreated or -resistant patients , and single-agent eribulin improves survival compared with the physician 's choice of treatment in patients treated previously with at least two regimens for MBC . Regardless of the regimen , proactive management to detect treatment-related adverse events in a timely manner remains important for ensuring effective delivery of treatment . Many promising investigational agents are in development , including DB05773 ( trastuzumab emtansine ) and pertuzumab for P04626 -positive disease , as well as P09874 ( poly [ adenosine diphosphate ribose ] polymerase-1 ) inhibitors and cetuximab for triple-negative disease . In addition , new options for the treatment of MBC following failure of an anthracycline and a taxane promise to improve patient outcomes . Nurses should remain vigilant for adverse events and remember that the goal of treatment remains control of the disease and palliation . Urinary pheromones promote P29323 /Akt phosphorylation , regeneration and survival of vomeronasal ( P30518 ) neurons . The G protein-coupled pheromone receptor neurons ( V1R and P30518 ) of the vomeronasal organ ( VNO ) are continually replaced throughout the lifetime of the mouse . Moreover , active signalling of V2Rs via the transient receptor potential 2(TRPC2) channel is necessary for regeneration of receptors , as the TRPC2 null mutant mouse showed a 75 % reduction of V2Rs by the age of two months . Here we describe P30518 mediated signalling in a neuronal line established from vomeronasal stem cells taken from postnatal female mice . Cells were immunoreactive for Galpha(o) and P30518 , whereas V1R and Galpha(i) immunoreactivity could not be detected . Biological ligands ( dilute urine and its protein fractions ) were found to increase proliferation and survival of these neurons . Dilute mouse urine but not artificial urine also induced P29323 , Akt and CREB signalling in a dose dependent way . The volatile fraction of male mouse urine alone was without effect while the fraction containing peptides ( > 5 kDa ) also stimulated P29323 and Akt phosphorylation . The P29323 , Akt and CREB phosphorylation response was sensitive to pertussis toxin , confirming the involvement of P30518 linked Galpha(o) . Dilute mouse urine or its high molecular weight protein fraction increased survival and proliferation of these neurons . Hence , urinary pheromones , which signal important social information via mature neurons , also promote survival and proliferation of their regenerating precursors . These data show that regenerating V2Rs respond to urine and the urinary peptides by activation of the Ras- P29323 and P19957 -Akt pathways , which appear to be important for vomeronasal neural survival and proliferation . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Proteomic profiling of cancer stem cells derived from primary tumors of P04626 /Neu transgenic mice . Human epidermal growth factor receptor 2 ( P04626 ) overexpression leads to mammary tumorigenesis and its elevated levels lead to increase in cancer stem cells ( CSCs ) , invasion , and metastasis . CSCs are resistant to radiation/chemotherapeutic drugs and are believed to be responsible for recurrence/relapse of cancer . CSCs are isolated using flow cytometry based sorting , although reliable , this technology hinders the convenient identification of molecular targets of CSCs . Therefore to understand the molecular players of increased CSC through P04626 overexpression and to develop meaningful targets for combination therapy , we isolated and characterized breast CSCs through convenient tumorsphere culture . We identified the altered protein expression in CSC as compared to non-CSC using LC-MS/MS and confirmed those results using qRT-PCR and Western blotting . P02794 1 ( P02794 ) was identified as a candidate gene , which is involved in iron metabolism and iron depletion significantly decreased the self-renewal of CSCs . We further performed in silico analysis of altered genes in tumorsphere and identified a set of genes ( P06454 , P26447 , P06703 , TNXRD1 , P23219 , P35354 , P02533 , and P02794 ) , representing possible molecular targets , which in combination showed a promise to be used as prognostic markers for breast cancer . DB05773 for P04626 -positive metastatic breast cancer . DB00072 emtansine in advanced human epidermal growth factor receptor 2-positive breast cancer . INTRODUCTION : DB00640 - trastuzumab emtansine ( DB05773 ) is a human epidermal growth factor receptor 2 ( P04626 ) -targeted antibody-drug conjugate composed of trastuzumab , a stable linker ( MCC ) , and the cytotoxic agent DM1 ( derivative of maytansine ; mertansine ) . DB05773 retains the mechanisms of action of trastuzumab , but also acts as a , selectively delivered , tubulin inhibitor . Following antigen-mediated binding to the tumor cell , DB05773 is endocytosed and intracellularly catabolized resulting in the release of its cytotoxic moiety . AREAS COVERED : DB05773 has completed Phase III development and compared favorably with the lapatinib/capecitabine combination with a superior response rate ( objective response rate [ ORR ] ) and duration of response , longer duration of disease control ( progression-free survival [ PFS ] ) , prolonged overall survival and improved tolerability and quality of life in patients with prior treatment with trastuzumab and a taxane . In a separate Phase III , DB05773 was compared with any other chosen regimen in patients who had at least received two prior P04626 -directed therapies . DB05773 nearly doubled PFS . EXPERT OPINION : DB05773 ( Kadcyla ) has become the treatment of choice in second-line and beyond for patients with advanced P04626 -expressing breast cancer . DB00072 emtansine : a novel antibody-drug conjugate for P04626 -positive breast cancer . DB00072 emtansine ( DB05773 ) is a novel P04626 -directed antibody-drug conjugate . DB05773 consists of the potent antimicrotubule agent DM1 , linked via a noncleavable linker to the P04626 -specific monoclonal antibody trastuzumab . Preclinical studies demonstrate that DB05773 has dual mechanisms of action : selective delivery of DM1 to the P04626 -positive ( P04626 (+) ) tumor cell combined with trastuzumab 's activation of antibody-dependent cell-mediated cytotoxicity and inhibition of P04626 -mediated signal transduction . In phase II studies , DB05773 was active in patients with trastuzumab- and lapatinib-refractory metastatic breast cancer and led to improved progression-free survival compared with the combination of trastuzumab and docetaxel in the first-line setting . In a recent phase III trial in patients with metastatic breast cancer who previously received trastuzumab and a taxane , DB05773 resulted in improved progression-free and overall survival compared with capecitabine and lapatinib . DB05773 is associated with a favorable toxicity profile ; reversible thrombocytopenia and hepatic transaminase elevations are the only grade ≥3 adverse event present in 5 % or more of patients . Alopecia , peripheral neuropathy , and neutropenia are distinctly uncommon . On the basis of its improved efficacy and toxicity compared with capecitabine/lapatinib , DB05773 should be considered the standard for patients with P04626 (+) metastatic breast cancer who have previously progressed on trastuzumab and a taxane . Results from additional randomized studies in metastatic breast cancer are pending , and trials in the (neo)adjuvant setting are being initiated . [ Chemotherapy for breast cancer refractory to anthracycline , taxane or trastuzumab ] . Anthracycline , taxane or trastuzumab play a central role in systemic chemotherapy for breast cancer . The standard of subsequent treatment is capecitabine , S-1 , vinorelbine , irinotecan or gemcitabine . DB04845 or nanoparticle paclitaxel is effective for taxane-resistant breast cancer . DB01259 proves effective for trastuzumab-resistant P04626 -overexpressing breast cancer and also for brain metastasis . DB05773 , pertuzumab and neratinib are promising drugs . In terms of antiangiogenic agents , bevacizumab in combination with taxane demonstrates efficacy . DB06626 , sunitinib or pazopanib is under investigation . It is necessary to study the best manner of sequence and combination in these drugs . P04626 -positive breast cancer : beyond trastuzumab . The outlook for patients with P04626 -positive breast cancer was revolutionized by the development of trastuzumab ( Herceptin ) , a humanized murine monoclonal antibody . Use of this agent led to improved overall survival when it was added to chemotherapy for the treatment of metastatic breast cancer . Improved understanding of mechanisms of resistance to trastuzumab has facilitated the development of novel agents for P04626 -positive breast cancer , and also resulted in superior outcomes when added to chemotherapy in the adjuvant setting . This review explores the use of several such agents , including lapatinib ( DB01259 ) , HSP90 inhibitors , DB05773 , and other tyrosine kinase inhibitors . Emerging data from trials of these agents indicate that the P04626 pathway remains a valid therapeutic target following disease progression on trastuzumab , and suggest a promising role for combined P04626 blockade with two or more agents . New therapies in P04626 -positive breast cancer : a major step towards a cure of the disease ? Overexpression of the human epidermal growth factor receptor 2 ( P04626 ) predicts a poor prognosis in metastatic breast cancer . While the introduction of P04626 -targeted therapies , such as the monoclonal antibody trastuzumab and the small-molecule tyrosine kinase inhibitor lapatinib , has significantly improved outcomes in P04626 + breast cancer compared with previously available therapies , use of these targeted therapies is often limited by the development of drug resistance and tolerability issues . These limitations create the need for further development and investigation of new targeted therapies that show potent and selective inhibition of these targets or closely connected molecular pathways . Recently , several agents have demonstrated promising activity in P04626 + metastatic breast cancer , either as monotherapy or in combination therapy , including the tyrosine-kinase inhibitors neratinib ( HKI-272 ) and afatinib ( BIBW-2992 ) and the anti- P04626 monoclonal antibodies pertuzumab and trastuzumab-DM1 ( DB05773 ) . Agents that target other molecular pathways , such as the vascular endothelial growth factor receptor , mammalian target of rapamycin , P19957 -kinases , insulin-like growth factor ( IGFR ) , HSP-90 , and other kinases also have potential , in combination with anti- P04626 and/or other systemic therapies , to be active in this subtype of breast cancer . Innovative clinical studies are required in well-characterized patient populations to define the true clinical value of these emerging new approaches . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits .

MH_train_41

MH_train_41

MH_train_41

interacts_with DB00530?

multiple_choice

Erlotinib in non-small cell lung cancer : a review . Erlotinib ( Tarceva , DB00530 ; Pfizer , Inc. ) is an orally-active , targeted inhibitor of the epidermal growth factor receptor ( P00533 / P00533 ) , which is part of a key regulatory pathway in cancer . Patients with advanced , incurable non-small cell lung cancer ( NSCLC ) may derive a clinical benefit from first- and second-line chemotherapy , but third-line treatment with available cytotoxic agents is not effective . Remarkably , P00533 / P00533 antagonists have demonstrated activity as second- and even third-line treatment for this disease . Erlotinib is the first of this novel class of drug to demonstrate a statistically significant and clinically relevant difference in overall survival , progression free survival and time to disease related symptoms ( cough , pain , shortness of breath ) compared with treatment with best supportive care in patients who have failed standard first- or second-line chemotherapy . This paper reviews the pharmacology , preclinical and clinical data to support the use of erlotinib in NSCLC . Targeting epidermal growth factor receptor : novel therapeutics in the management of cancer . Overexpression of epidermal growth factor receptor ( P00533 ) in epithelial tumors , including head and neck , lung , breast , colon and other solid tumors , has frequently been correlated with poor prognosis , thus stimulating efforts to develop new cancer therapies that target P00533 . Monoclonal antibodies and tyrosine kinase inhibitors specifically targeting P00533 are the most well-studied and hold substantial promise of success . Several compounds of monoclonal antibodies and tyrosine kinase inhibitors targeting P00533 have been studied and clinical trials are now underway to test the safety and efficacy of these targeting strategies in several human tumors . This review will address each of these agents alone or in combination with radiation or chemotherapy and highlight some of these promising developments . Cetuximab ( Erbitux ) is being evaluated in combination with radiation or chemotherapy in Phase III trials . Other compounds such as h-R3 , DB01269 , P50402 -55900 and ICR-62 have proved to be effective in targeting malignant cells alone or in combination with traditional therapies . Tyrosine kinase inhibitors targeting the intracellular domain of P00533 , including ZD-1839 ( gefitinib , DB00317 ) , DB00530 ( Erlotinib/Tarceva ) , PD-153053 , PD-168393 and DB05424 , have been studied in clinical setting alone or in combination with radiation or chemotherapy . ZD-1839 is being studied in a Phase III trial in patients with advanced non-small cell lung cancer . P00533 targeted treatment by monoclonal antibodies and tyrosine kinase inhibitors have been proven to sensitize tumor cells to the effects of chemotherapy and radiation therapy . The synergistic activities and nonoverlapping toxicities of these compounds allow concomitant administration with cytotoxic therapy . Challenges of evaluating P00533 targeted agents exist in selecting the optimal dosages and determining long-term toxicity . Soothing the watchman : telomerase reduces the p53-dependent cellular stress response . In addition to conferring an indefinite replicative life span , telomerase renders p16(-) human mammary epithelial cells ( HMEC ) resistant to growth arrest by TGFbeta or by loss of P01133 or insulin signaling . In contrast to earlier reports , we recently found that growth factor signaling was not directly affected by telomerase expression . Rather , short dysfunctional or near-dysfunctional telomeres in proliferating telomerase(-) HMEC sensitized the cells to p53-dependent signals for growth arrest . We showed that during serial passage and before any signs of replicative senescence , HMEC lacking telomerase experience enhanced p53 stability and DNA damage signaling , as determined by increased phosphorylation on p53-Ser15 and Chk2-Thr68 , and formation of Q12888 /phosphorylated histone P16104 foci at chromosome ends . This heightened activity of the p53 pathway enhanced the efficiency with which cells arrested growth in response to TGFbeta or to P01133 or insulin withdrawal , and was abolished by ectopic expression of hTERT , the catalytic subunit of telomerase . Telomerase elongated short telomeres , thereby reducing the basal level of activated p53 and raising cellular tolerance for other p53-dependent signals , including those emanating from non-genotoxic sources . These findings explain a number of observed effects of telomerase expression on cell growth and survival without postulating additional functions for telomerase . 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 . Interstitial lung disease in patients with non-small-cell lung cancer treated with epidermal growth factor receptor inhibitors . Interstitial lung disease ( ILD ) refers to a diverse range of pulmonary fibrotic disorders and may be hard to accurately diagnose , as distinguishing it from other pulmonary diseases can be difficult . Estimations of the incidence in populations are confounded by the complexity of the different forms of the disorder . In addition , ILD is a comorbid disease of lung cancer and is seen after most forms of chemotherapy and radiotherapy for advanced lung cancer . Incidences of > or=10 % have been reported ; however , whatever the true incidence , both chemotherapy and radiotherapy enhance the risk of developing ILD . ILD has also been reported with the epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors , including erlotinib ( Tarceva , DB00530 ) and gefitinib ( IRESSA ) . In a large number of gefitinib-treated patients ( n > 185,000 ) an incidence of approx 1 % has been observed ( approx 2 % in Japan ; 0.3 % in the rest of the world ) . Nevertheless , as with other treatments for advanced non-small-cell lung cancer , the clinical benefit outweighs the risk of ILD . In this article , we review the data on ILD with P00533 inhibitors and other common lung cancer treatments . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . 5- Q13049 receptor induces P29323 phosphorylation and proliferation through ADAM-17 tumor necrosis factor-alpha-converting enzyme ( P78536 ) activation and heparin-bound epidermal growth factor-like growth factor ( HB- P01133 ) shedding in mesangial cells . In this study , we present multiple lines of evidence to support a critical role for heparin-bound P01133 ( epidermal growth factor ) -like growth factor ( HB- P01133 ) and tumor necrosis factor-alpha-converting enzyme ( P78536 ) ( P78536 ) in the transactivation of P01133 receptor ( P00533 ) , P29323 phosphorylation , and cellular proliferation induced by the 5-HT(2A) receptor in renal mesangial cells . 5-hydroxy-tryptamine ( 5-HT ) resulted in rapid activation of P78536 , HB- P01133 shedding , P00533 activation , P29323 phosphorylation , and longer term increases in DNA content in mesangial cells . P29323 phosphorylation was attenuated by 1 ) neutralizing P00533 antibodies and the P00533 kinase inhibitor , AG1478 , 2 ) neutralizing HB- P01133 , but not amphiregulin , antibodies , heparin , or CM197 , and 3 ) pharmacological inhibitors of matrix-degrading metalloproteinases or P78536 small interfering RNA . Exogenously administered HB- P01133 stimulated P29323 phosphorylation . Additionally , P78536 was co-immunoprecipitated with HB- P01133 . Small interfering RNA against P78536 also blocked 5-HT-induced increases in P29323 phosphorylation , HB- P01133 shedding , and DNA content . In aggregate , this work supports a pathway map that can be depicted as follows : 5-HT --> 5-HT(2A) receptor --> P78536 --> HB- P01133 shedding --> P00533 --> P29323 --> increased DNA content . To our knowledge , this is the first time that P78536 has been implicated in 5-HT-induced P00533 transactivation or in proliferation induced by a G protein-coupled receptor in native cells in culture . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Erlotinib ( DB00530 ) -induced inhibition of transitional cell carcinoma of bladder cell line growth is enhanced by interferon-alpha . OBJECTIVE : To examine whether erlotinib gives similar results to gefitinib , a small molecule epidermal growth factor receptor ( P00533 / P00533 ) tyrosine kinase ( TK ) inhibitor that inhibits the growth of human bladder cancer cell lines in vitro , and given that interferon-alpha ( IFNalpha ) promotes an antiproliferative effect of P00533 / P00533 inhibitors on colon cancer cell lines , to also determine the effects of erlotinib alone or together with INFalpha on bladder cancer cell lines , and whether sensitivity is influenced by P00533 / P00533 mutation status . MATERIALS AND METHODS : Seven bladder cancer cell lines were characterized for P00533 / P00533 expression , then treated with erlotinib alone , IFNalpha alone , or IFNalpha plus erlotinib . Cell growth inhibition was assessed by crystal-violet staining and P00533 / P00533 expression by flow cytometry . Synergy was evaluated using the combination index of Chou and Talalay . DNA from these cell lines in the linear growth phase and from 14 bladder cancer tissue samples were tested for P00533 /EGFRTK mutations . RESULTS : Cell-surface P00533 / P00533 expression was present in all seven bladder cancer cell lines . Both erlotinib and IFNalpha independently were significantly antiproliferative , and combined treatment synergistically enhanced the sensitivity in six of the seven cell lines . No bladder cancer cell lines or tissues tested expressed P00533 /EGFRTK mutations . CONCLUSION : Erlotinib inhibits the growth of human bladder cancer cell lines . Enhanced inhibition in the presence of IFNalpha is not determined by the presence of P00533 /EGFRTK mutations . This study might have clinical implications for improving the treatment of bladder cancer . Combined effects of C225 and 125-iodine seed radiation on colorectal cancer cells . BACKGROUND : To characterize the effect of combined treatment of the anti-epidermal growth factor receptor ( P00533 ) monoclonal antibody C225 and 125-iodine ( 125I ) seed radiation in human colorectal cancer . METHODS : We treated LS180 cells with 125I continuous low dose rate radiation in the presence and absence of 100 nM C225 . The clonogenic capacity , cellular proliferation , cell cycle distribution , apoptosis , and molecular pathways of the cells following the treatments were analyzed in vitro . RESULTS : The sensitizer enhancement ratio of C225 was approximately 1.4 . Treatment with C225 and radiation alone produced significant inhibition of cell growth , but combination therapy produced greater inhibition than either treatment administered alone . C225 increased the radiation-induced apoptosis and the fraction of γ- P16104 foci positive cells at 48 h after treatment . The Akt phosphorylation level was lower in the cells receiving the combination treatment than in the cells treated with radiation or C225 alone . CONCLUSIONS : These findings indicate that C225 sensitizes LS180 cells to 125I seed radiation . Growth inhibition is mediated by inducing apoptosis and not cell cycle arrest . Additionally , we confirmed that C225 impairs DNA repair by reducing the cellular level of the P78527 and P12956 proteins . Furthermore , the inhibition of Akt signaling activation may be responsible for the C225-mediated radiosensitization . Rationale for investigation of epidermal growth factor receptor inhibitors in definitive treatment of locally advanced non-small cell lung cancer and head and neck cancer . Designing targeted therapies has become an important field in cancer therapeutics . The epidermal growth factor receptor ( P00533 ) is a molecular target that has gained immense attention as preclinical and clinical studies have supported its potential role for therapy of a variety of cancers , including non-small cell lung cancer ( NSCLC ) and head and neck ( HN ) cancer . Several compounds that specifically inhibit P00533 have been developed , including ZD1839 , C225 , and DB00530 . Interestingly , studies suggesting a potential role for P00533 inhibitors as an adjunct to the current combined-modality approach for therapy of NSCLC and HN cancer have been performed in the preclinical and clinical setting . Therefore , determining the potential of P00533 inhibitors to improve the efficacy of standard combined-modality regimens ( chemotherapy/radiation therapy +/- surgery ) for NSCLC and HN cancer patients is of the utmost importance . An overview of the rationale and the ongoing/proposed studies aimed at determining the role for P00533 inhibitors in combination with radiation therapy for NSCLC and HN cancer patients will be presented . P04626 up-regulates P26447 and several other prometastatic genes in medulloblastoma . Medulloblastoma is frequently disseminated throughout the central nervous system by the time of diagnosis . Conventional therapeutic approaches have not reduced the high mortality associated with metastatic medulloblastoma and little is known regarding the molecular mechanisms that promote tumor invasion . Previously , we reported that overexpression of P04626 in medulloblastoma is associated with poor prognosis and metastasis . Here , we demonstrate that P04626 overexpression increases the migration of medulloblastoma cells across basement membranes in vitro . Furthermore , using microarray expression profiling , we show that P04626 up-regulates the expression of prometastatic genes in medulloblastoma cells . These include P26447 , which was previously shown to promote metastasis of breast cancer . We demonstrate that P26447 is a direct target of P04626 signaling in medulloblastoma cells via a pathway involving phosphatidylinositol 3-kinase , P31749 , and extracellular signal-regulated kinase 1/2 and that levels of P04626 and P26447 are tightly correlated in samples of primary medulloblastoma . Finally , we show that P04626 -dependent medulloblastoma cell invasion in vitro and prometastatic gene expression in vivo can be blocked using the P00533 tyrosine kinase inhibitor DB00530 . These data identify an P04626 driven prometastatic pathway that may provide a novel target for therapeutic intervention in metastatic medulloblastoma . Beyond the TRIBUTE trial : integrating P00533 / P00533 tyrosine kinase inhibitors with chemotherapy in advanced NSCLC . Evaluation of : Herbst RS , Prager D , Hermann R et al. : TRIBUTE : a Phase III trial of erlotinib hydrochloride ( DB00530 ) combined with carboplatin and paclitaxel chemotherapy in advanced non-small cell lung cancer . J . Clin . Oncol . 23 , 5892-5899 ( 2005 ) . Patients diagnosed with advanced non-small cell lung cancer have limited therapeutic options . A large , randomized , controlled trial of the human epidermal growth factor receptor tyrosine-kinase inhibitor , erlotinib ( Tarceva ) , plus standard first-line chemotherapy did not meet its primary end point of improved survival in the overall population , but did reveal a striking survival benefit in a subset of patients who had never smoked . There are a number of possible explanations for the lack of overall benefit , including the use of an unselected patient population , the need for alternatives to concurrent administration , and a postulated pathophysiological interaction between erlotinib and chemotherapy . Ongoing studies investigating alternative schedules and sequences of administration with chemotherapy will help clinicians to determine how agents such as erlotinib can best be combined with standard cytotoxic agents . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Targeting the P00533 pathway for cancer therapy . Clinical studies have shown that HER-2/Neu is over-expressed in up to one-third of patients with a variety of cancers , including B-cell acute lymphoblastic leukemia ( B-ALL ) , breast cancer and lung cancer , and that these patients are frequently resistant to conventional chemo-therapies . Additionally , in most patients with multiple myeloma , the malignant cells over-express a number of epidermal growth factor receptors ( P00533 )s and their ligands , HB- P01133 and amphiregulin , thus this growth-factor family may be an important aspect in the patho-biology of this disease . These and other , related findings have provided the rationale for the targeting of the components of the P00533 signaling pathways for cancer therapy . Below we discuss various aspects of P00533 -targeted therapies mainly in hematologic malignancies , lung cancer and breast cancer . Beside novel therapeutic approaches , we also discuss specific side effects associated with the therapeutic inhibition of components of the P00533 -pathways . Alongside small inhibitors , such as DB01259 ( DB01259 , GW572016 ) , Gefitinib ( DB00317 , ZD1839 ) , and Erlotinib ( Tarceva , DB00530 ) , a significant part of the review is also dedicated to therapeutic antibodies ( e.g. : DB00072 /Herceptin , DB06366 / DB06366 /rhuMab-2C4 , Cetuximab/Erbitux/IMC-C225 , DB01269 /Abenix/ DB01269 , and also DB05294 ) . In addition , we summarize , both current therapy development driven by antibody-based targeting of the P00533 -dependent signaling pathways , and furthermore , we provide a background on the history and the development of therapeutic antibodies . 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 . The emerging role of epidermal growth factor receptor inhibitors in ovarian cancer . P00533 ( P00533 ) inhibitors are a new biologically targeted therapy , which may offer new hope in the treatment of patients with advanced or recurrent ovarian cancers . In this review , we summarize and discuss the results of research to date on P00533 inhibitors with particular emphasis on ovarian cancer . We reviewed data identified by searches of MEDLINE , PubMed , and abstracts from the proceedings of the American Society of Clinical Oncology meetings from 1998 to 2006 , with the search terms " Ovarian Cancer, " " P00533 , " " gefitinib , ZD1839 , DB00317 , " " erlotinib , DB00530 , Tarceva, " " DB05424 , " " DB01259 , lapatinib, " " PKI-166, " " Q9Y259 569, " " anti- P00533 antibodies, " " trastuzumab , Herceptin, " " cetuximab , Erbitux , IMC-C225, " " matuzumab , P50402 72000, " " panitumamab , DB01269 , " " pertuzumab , " and " vandetanib , rINN , DB05294 , DB05294 . " Phase II trials of both small molecule inhibitors of P00533 - and antibody-based inhibitors are currently ongoing in ovarian cancer and emerging data suggest that their activity in unselected women with advanced or recurrent ovarian cancer is modest , when utilized as a single agent . It is possible that these agents will be highly effective in smaller subsets of patients whose tumors are dependent on P00533 signaling , perhaps through activating mutations in P00533 or its downstream pathway . Targeted therapy with P00533 inhibitors is an untapped potential resource in the treatment of advanced or recurrent ovarian cancer . Ongoing trials will elucidate the most effective strategies to use these agents individually or in combination with traditional chemotherapeutic agents . Nimotuzumab suppresses epithelial-mesenchymal transition and enhances apoptosis in low-dose UV-C treated salivary adenoid cystic carcinoma cell lines in vitro . Salivary adenoid cystic carcinoma ( SACC ) , which is one of the most common malignant tumors of the salivary glands , is associated with a poor long-term outcome . There are currently few therapeutic options for patients with SACC . Recent studies have shown the potential of the application of ultraviolet-C ( UV-C ) irradiation for the treatment of human cancer . In the present study , we investigated the effects of UV-C in the SACC cell lines SACC-83 and SACC-LM . High-dose UV-C ( 200 J/m ) induced apoptosis and inhibited colony formation significantly . However , low-dose UV-C ( 10 J/m ) , which had little effect on apoptosis and colony formation , increased the ability of migration in SACC cells accompanied by a decrease in P12830 and an increase in vimentin , suggesting the occurrence of epithelial-mesenchymal transition ( EMT ) . Low-dose UV-C ( 10 J/m ) also resulted in upregulation of the phosphorylated forms of epidermal growth factor receptor ( P00533 ) and Akt ( p- P00533 and p-Akt , respectively ) . Pretreatment with Nimotuzumab , an anti- P00533 monoclonal antibody , reversed the EMT as well as upregulation of p- P00533 /p-Akt induced by UV-C . Moreover , Nimotuzumab enhanced UV-C induced apoptosis and inhibition of colony formation . Our results indicate that EMT exerts a protective effect against apoptosis induced by low-dose UV-C . Thus , the combined application of Nimotuzumab and low-dose UV-C in vitro has an advantageous antitumor effect in SACC compared with the application of UV-C alone . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Phase II study of erlotinib ( DB00530 ) in patients with metastatic colorectal cancer . Erlotinib ( Tarceva , DB00530 ) , a potent epidermal growth factor receptor tyrosine kinase inhibitor ( P00533 ) , was evaluated in a phase II study to assess its activity in patients with metastatic colorectal cancer . In all , 38 patients with metastatic colorectal cancer were treated with erlotinib at a continuous daily oral dose of 150 mg . Radiological evaluation was carried out every 8 weeks and tumour biopsies were performed before treatment and on day 8 . Of 31 evaluable patients , 19 ( 61 % ) had progressive disease and 12 ( 39 % ) had stable disease ( s.d. ) . The median time to progression for those patients having s.d. was 123 days ( range 108-329 days ) . The most common adverse events were rash in 34 patients and diarrhoea in 23 patients . Correlative studies were conducted to investigate the effect of erlotinib on downstream signalling . Tumour tissue correlations were based on usable tissue from eight match paired tumour samples pre- and on therapy , and showed a statistically significant decrease in the median intensity of both pEGFR ( P=0.008 ) and phospho-extracellular signal-regulated kinase ( P29323 ) ( P=0.008 ) a week after commencement of treatment . No other statistically significant change in tumour markers was observed . Erlotinib was well tolerated with the most common toxicities being rash and diarrhoea . More than one-third of evaluable patients had s.d. for a minimum of 8 weeks . Correlative studies showed a reduction in phosphorylated P00533 and P29323 in tumour tissue post-treatment . Tamoxifen enhances erlotinib-induced cytotoxicity through down-regulating AKT-mediated thymidine phosphorylase expression in human non-small-cell lung cancer cells . Tamoxifen is a triphenylethylene nonsteroidal estrogen receptor ( ER ) antagonist used worldwide as an adjuvant hormone therapeutic agent in the treatment of breast cancer . However , the molecular mechanism of tamoxifen-induced cytotoxicity in non-small cell lung cancer ( NSCLC ) cells has not been identified . P19971 ( TP ) is an enzyme of the pyrimidine salvage pathway which is upregulated in cancers . In this study , tamoxifen treatment inhibited cell survival in two NSCLC cells , H520 and H1975 . Treatment with tamoxifen decreased TP mRNA and protein levels through AKT inactivation . Furthermore , expression of constitutively active AKT ( AKT-CA ) vectors significantly rescued the decreased TP protein and mRNA levels in tamoxifen-treated NSCLC cells . In contrast , combination treatment with PI3K inhibitors ( LY294002 or wortmannin ) and tamoxifen further decreased the TP expression and cell viability of NSCLC cells . Knocking down TP expression by transfection with small interfering RNA of TP enhanced the cytotoxicity and cell growth inhibition of tamoxifen . Erlotinib ( Tarceva , DB00530 ) , an orally available small molecular inhibitor of epidermal growth factor receptor ( P00533 ) tyrosine kinase , is approved for clinical treatment of NSCLC . Compared to a single agent alone , tamoxifen combined with erlotinib resulted in cytotoxicity and cell growth inhibition synergistically in NSCLC cells , accompanied with reduced activation of phospho-AKT and phospho- P27361 /2 , and reduced TP protein levels . These findings may have implications for the rational design of future drug regimens incorporating tamoxifen and erlotinib for the treatment of NSCLC . Somatic mutations of the epidermal growth factor receptor and non-small-cell lung cancer . Frequent overexpression of epidermal growth factor receptor ( P00533 ) in non-small-cell lung cancer ( NSCLC ) makes P00533 a new therapeutic target . Two specific P00533 tyrosine kinase inhibitors , gefitinib ( ZD1839 , DB00317 ) and erlotinib ( DB00530 , Tarceva ) , have been developed and approved by the US Food and Drug Administration for second-line and third-line treatment of advanced NSCLC . Clinical trials have shown considerable variability in the response rate between different patients with NSCLC , which led to the discovery of somatic P00533 -activating mutations . This brief review summarises the discovery and functional consequences of the mutations , their clinicopathological features and significant implications in the treatment and prognosis of NSCLC . Extracellular signal-regulated kinase and the small GTP-binding protein , Rac , contribute to the effects of transforming growth factor-beta1 on gene expression . The kinases and regulatory proteins that convey signals initiated by transforming growth factor-beta ( TGF-beta ) to the nucleus are poorly characterized . To study the role of the extracellular signal-regulated kinase ( P29323 ) pathway in this process , we transiently transfected NIH 3T3 fibroblasts with TGF-beta-responsive luciferase reporter genes and expression vectors designed to interrupt this kinase cascade . Mitogen-activated protein ( Q96HU1 ) kinase phosphatase-1 and a dominant negative Q96HU1 / P29323 kinase 1 mutant reduced stimulation of plasminogen activator inhibitor-1 ( P05121 ) promoter activity by TGF-beta1 from 11.5- to 4-fold and 4.9-fold , respectively . Similar results were observed with the type I collagen promoters . TGF-beta1 increased P27361 activity 4.5-fold at 5 min and 3 . 1-fold at 3 h , while Jun kinase and p38 activity were not affected . Cotransfection of a dominant negative mutant of the small G protein , Rac , but not dominant negative Ras , Cdc42 , or Rho mutants , reduced the effects of TGF-beta1 on the P05121 promoter by approximately half . In support of a role for Rac in signaling by TGF-beta , GTP binding to Rac was increased 3.7-fold following exposure of NIH 3T3 cells to TGF-beta1 for 3 min . These findings indicate that TGF-beta1 modulates gene expression partly through P29323 and Rac in NIH 3T3 cells . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Overview of tyrosine kinase inhibitors in clinical breast cancer . Studies of cell models and profiling of clinical breast cancer material to reveal the mechanisms of resistance to anti-oestrogen therapy , and to tamoxifen in particular , have reported that this phenomenon can be associated with increased expression and signalling through erbB Type 1 growth factor receptors , notably the epidermal growth factor receptor ( P00533 ) and P04626 . Further molecular studies have revealed an intricate interlinking between such growth factor receptor pathways and oestrogen receptor ( ER ) signalling . Inhibition of receptor tyrosine kinase activity involved in the P00533 signalling cascade forms the basis for the use of P00533 specific tyrosine kinase inhibitors exemplified by gefitinib ( ZD1839 , DB00317 ) and erlotinib ( DB00530 , Tarceva ) . Such agents have proved promising in pre-clinical studies and are currently in clinical trials in breast cancer , where gefitinib has been studied more extensively to date . Here , we present an overview of the current development of gefitinib in clinical breast cancer . This includes results from our clinical breast cancer trial 1839IL/0057 that demonstrate the efficacy of gefitinib within ER-positive , tamoxifen-resistant patients with locally advanced/metastatic disease , where parallel decreases in P00533 signal transduction and the Ki67 ( Q86YT6 ) proliferation marker can be detected as predicted from model system studies . We also consider trials examining combination treatment with gefitinib and anti-hormonal strategies that will begin to address the clinically important question of whether gefitinib can delay/prevent onset of anti-hormone resistance . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Epithelial growth factor receptor interacting agents . The data reviewed here have further established the promise of anti- P00533 -targeted therapies . This statement is supported by the evidence of antitumor activity of the TK inhibitors ZD1839 and DB00530 against several tumor types and by the ability of the monoclonal antibody IMC-C225 to reverse clinical chemotherapy resistance . These results are further supported by an emerging number of compounds , monoclonal antibodies , and TK inhibitors directed at the P00533 that are in clinical development ( see Fig. 2 , Table 1 ) . Among the TK inhibitors , these compounds can be further categorized by their receptor specificity and reversibility of binding . In the case of anti- P00533 monoclonal antibodies , compounds in clinical development include chimeric , humanized , and bispecific antibodies . The fundamental observation is that these compounds have shown activity in several tumor types , including NSCL cancer , prostate carcinoma , colorectal carcinoma , ovarian carcinoma , renal cell carcinoma , and head and neck cancers . These findings observed with different agents and in different tumor types validate P00533 as a target for cancer therapy . The results of ongoing studies with these agents in diverse indications and tumor types may establish the role of these promising therapies to our current cancer treatments . Local immunotherapy of glioma patients with a combination of 2 bispecific antibody fragments and resting autologous lymphocytes : evidence for in situ t-cell activation and therapeutic efficacy . After adoptive transfer of pre-activated lymphocytes into the operation cavity of glioma patients , tumor regression and improved survival have been reported in some patients . Results were most impressive when bispecific antibodies with tumor x CD3 specificity were also applied . In this study , we attempted to avoid time-consuming pre-activation procedures for adoptively transferred cells by using a combination of bispecific antibodies directed to the P01133 receptor ( P00533 ) on tumor cells and to CD3 and P10747 on T cells . Eleven patients with high-grade malignant glioma received 3 injections of 2 bispecific antibody fragments ( P00533 x CD3 and P00533 x P10747 ) together with freshly isolated autologous lymphocytes via an Ommaya reservoir . Intracavitary fluid aspirated during immunotherapy was examined for markers of T-cell activation . Increased levels of soluble P60568 receptor and P01375 were detected in the intracavitary fluid of all patients tested . Two of the 11 treated patients experienced a beneficial response to therapy as defined by a transient contrast enhancement in subsequent Q9BWK5 scans and prolonged survival . Side effects were transient and consisted of fever , nausea , headache and aggravation of pre-existing neurologic deficits . These adverse effects were most likely due to the antibody construct containing anti-CD3 specificity . Two patients developed cerebral edema and required steroid treatment . Attenuated P08908 receptor signaling in brains of suicide victims : involvement of adenylyl cyclase , phosphatidylinositol 3-kinase , Akt and mitogen-activated protein kinase . Positron emission tomography studies in major depression show reduced serotonin (5-HT)1A receptor antagonist-binding potentials in many brain regions including occipital cortex . The functional meaning of this observation in terms of signal transduction is unknown . We used postmortem brain samples from depressed suicide victims to examine the downstream effectors of P08908 receptor activation . The diagnosis was established by means of psychological autopsy using Diagnostic and Statistical Manual of Mental Disorders ( DSM ) III-R criteria . Measurements of [35S]GTPgammaS binding to Galphai/o in the occipital cortex of suicide victims and matched controls revealed a blunted response in suicide subjects and a decrease in the coupling of P08908 receptor to adenylyl cyclase . No significant group differences were detected in the expression levels of Galphai/o , Galphaq/11 or Galphas proteins , or in the activity of DB02527 -dependent protein kinase A . Studies of a parallel transduction pathway downstream from P08908 receptor activation demonstrated a decrease in the activity of phosphatidylinositol 3-kinase and its downstream effector Akt , as well as an increase in P60484 ( phosphatase and tensin homolog deleted on chromosome 10 ) , the phosphatase that hydrolyzes phosphatidylinositol 3,4,5-triphosphate . Finally , the activation of extracellular signal-regulated kinases 1 and 2 was attenuated in suicide victims . These data suggest that the alterations in agonist-stimulated P08908 receptor activation in depressed suicide victims are also manifest downstream from the associated G protein , affecting the activity of second messengers in two P08908 receptor transduction pathways that may have implications for cell survival . Rationale and clinical validation of epidermal growth factor receptor as a target in the treatment of head and neck cancer . Recurrent/metastatic head and neck cancer is an area of high , unmet treatment need . There is a strong rationale for targeting the epidermal growth factor receptor ( P00533 ) in head and neck cancer as most of these tumors express high levels of P00533 relative to normal tissue , with high expression correlating with poor patient outcome . This rationale has been validated in extensive preclinical studies . Two small molecules with P00533 inhibitory activity , gefitinib ( ' DB00317 ' , ZD1839 ) and erlotinib ( ' Tarceva ' , DB00530 ) , and a humanized monoclonal antibody against the P00533 extracellular domain , cetuximab ( ' Erbitux ' , C225 ) , are in clinical trials for advanced head and neck cancer . The initial results of these trials are promising . Gefitinib and erlotinib show activity as monotherapy in patients with recurrent or metastatic head and neck cancer , and have an acceptable safety profile compared with conventional chemotherapy . Gefitinib , which can be given at doses below the maximum tolerated dose , is associated with slightly lower rates of adverse events than erlotinib , which is dosed at the maximum tolerated dose . Combinations of cetuximab with radiotherapy or platinum-based chemotherapy have also shown activity in phase I/II studies . Both gefitinib and cetuximab have entered phase III studies . The results of these trials , which will mature over the next few years , will help determine the optimal use of P00533 agents in head and neck cancers . P00533 as a therapeutic target in colorectal cancer . The epidermal growth factor receptor ( P00533 ) is widely expressed in advanced colorectal cancers ( CRCs ) , and higher levels of P00533 are inversely related to survival in these patients . Two general strategies have been used to block P00533 signaling : preventing ligand binding with anti- P00533 monoclonal antibodies ( eg , cetuximab and DB01269 ) and inhibiting its intrinsic tyrosine kinase with small molecules ( eg , gefitinib [ DB00317 ] and erlotinib [ DB00530 ,Tarceva ] ) . Phase II trials of cetuximab suggest that it might be an effective treatment option alone or in combination with standard therapies as first- or second-line therapy . Phase I studies evaluating other P00533 inhibitors in patients with CRC have been reported . The inclusion of anti- P00533 therapies into standard treatment is the subject of current clinical trials . Combination treatment of glioblastoma multiforme cell lines with the anti-malarial artesunate and the epidermal growth factor receptor tyrosine kinase inhibitor DB00530 . New drugs and combination modalities for otherwise non-responsive brain tumors are urgently required . The anti-malarial artesunate ( O00253 ) and the P00533 tyrosine kinase inhibitor DB00530 reveal profound cytotoxic activity . The effectiveness of a combination treatment and the underlying molecular determinants of cellular response are unknown . In the present investigation , we studied O00253 and DB00530 in glioblastoma multiforme ( GBM ) cell lines . Supra-additive inhibition of cell growth was observed in U-87MG.DeltaEGFR cells transduced with a deletion-mutant constitutively active P00533 gene , while additive effects were present in cells transduced with wild-type P00533 ( U-87MG.WT-2N ) , kinase-deficient P00533 ( U-87MG.DK-2N ) , mock vector controls ( U-87MG.LUX ) , or non-transduced parental U-87MG cells . Among nine other non-transduced GBM cell lines , supra-additive effects were found in two cell lines ( G-210GM , G-599GM ) , while O00253 and DB00530 acted in an additive manner in the other seven cell lines ( G-211GM , G-750GM , G-1163GM , G-1187GM , G-1265GM , G-1301GM , and G-1408GM ) . Sub-additive or antagonistic effects were not observed . Genomic gains and losses of genetic material in the non-transduced cell lines as assessed by comparative genomic hybridization were correlated with the IC(50) values for O00253 and DB00530 and subsequently subjected to hierarchical cluster analysis and cluster image mapping . A genomic profile of imbalances was detected that predicted cellular response to O00253 and DB00530 . The genes located at the genomic imbalances of interest may serve as candidate resistance genes of GBM cells towards O00253 and DB00530 . In conclusion , the combination treatment of O00253 and DB00530 resulted in an increased growth inhibition of GBM cell lines as compared to each drug alone . Erlotinib ( Tarceva , DB00530 ) antagonizes DB00171 -binding cassette subfamily B member 1 and DB00171 -binding cassette subfamily G member 2-mediated drug resistance . It has been reported that gefitinib , an epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor ( TKI ) , has the ability to modulate the function of certain DB00171 -binding cassette ( DB01048 ) transporters and to reverse ABC subfamily B member 1 ( P08183 ; P-glycoprotein ) - and ABC subfamily G member 2 ( Q9UNQ0 ; breast cancer resistance protein/mitoxantrone resistance protein ) -mediated multidrug resistance ( MDR ) in cancer cells . However , it is unknown whether other P00533 TKIs have effects similar to that of gefitinib . In the present study , we have investigated the interaction of another P00533 TKI , erlotinib , with selected ABC drug transporters . Our findings show that erlotinib significantly potentiated the sensitivity of established P08183 or Q9UNQ0 substrates and increased the accumulation of paclitaxel or mitoxantrone in P08183 - or Q9UNQ0 -overexpressing cells . Furthermore , erlotinib did not significantly alter the sensitivity of non- P08183 or non- Q9UNQ0 substrates in all cells and was unable to reverse MRP1-mediated MDR and had no effect on the parental cells . However , erlotinib remarkably inhibited the transport of E(2)17 beta G and methotrexate by Q9UNQ0 . In addition , the results of ATPase assays show that erlotinib stimulated the ATPase activity of both P08183 and Q9UNQ0 . Interestingly , erlotinib slightly inhibited the photolabeling of P08183 with [(125)I]iodoarylazidoprazosin ( IAAP ) at high concentration , but it did not inhibit the photolabeling of Q9UNQ0 with IAAP . Overall , we conclude that erlotinib reverses P08183 - and Q9UNQ0 -mediated MDR in cancer cells through direct inhibition of the drug efflux function of P08183 and Q9UNQ0 . These findings may be useful for cancer combinational therapy with erlotinib in the clinic . Phase II study of carboplatin and erlotinib ( Tarceva , DB00530 ) in patients with recurrent glioblastoma . Targeting the epidermal growth factor receptor ( P00533 ) may be effective in a subset of glioblastoma patients . This phase II study assessed the clinical activity of erlotinib plus carboplatin and to determine molecular predictors of response . The primary endpoint was progression free survival ( PFS ) . Patients with recurrent glioblastoma with no more than two prior relapses received carboplatin intravenously on day 1 of every 28-day cycle ( target AUC of 6 mg x ml/min ) . Daily erlotinib at 150 mg/day was dose escalated to 200 mg/day , as tolerated . Clinical and Q9BWK5 assessments were made every 4 and 8 weeks , respectively . Tumor tissue was evaluated for P00533 , AKT and phosphatase and tensin homolog ( P60484 ) status . One partial response ( PR ) was observed out of 43 assessable patients . Twenty patients ( 47 % ) had stable disease ( SD ) for an average of 12 weeks . Median PFS was 9 weeks . The 6-month PFS rate was 14 % . Median overall survival ( OS ) was 30 weeks . This regimen was well tolerated with grade 3/4 toxicities of fatigue , leukopenia , thrombocytopenia and rash requiring dose reductions . A recursive partitioning analysis ( RPA ) predicted that patients with KPS > or=90 treated with more than 1 prior regimen had the highest OS . No correlation was observed between P00533 , Akt or P60484 expression and either PFS or OS . Carboplatin plus erlotinib is well tolerated but has modest activity in unselected patients . Future trials should be stratified based on optimal molecular or clinical characteristics . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma . Tumor-derived exosomes contain biologically active proteins and messenger and microRNAs ( miRNAs ) . These particles serve as vehicles of intercellular communication and are emerging mediators of tumorigenesis and immune escape . Here , we isolated 30-100 nm exosomes from the serum of patients with nasopharyngeal carcinoma ( NPC ) or the supernatant of TW03 cells . Increased circulating exosome concentrations were correlated with advanced lymphoid node stage and poor prognosis in NPC patients ( P < 0.05 ) . TW03-derived exosomes impaired T-cell function by inhibiting T-cell proliferation and Th1 and Th17 differentiation and promoting Treg induction by NPC cells in vitro . These results are associated with decreases in P29323 , P42224 , and P40763 phosphorylation and increases in P42229 phosphorylation in exosome-stimulated T-cells . TW03-derived exosomes increased the proinflammatory cytokines IL-1β , P05231 , and P22301 but decreased IFNγ , P60568 , and Q16552 release from P01730 + or CD8+ T-cells . Furthermore , five commonly over-expressed miRNAs were identified in the exosomes from patient sera or NPC cells : hsa-miR-24-3p , hsa-miR-891a , hsa-miR-106a-5p , hsa-miR-20a-5p , and hsa-miR-1908 . These over-expressed miRNA clusters down-regulated the Q9P0L2 signaling pathway to alter cell proliferation and differentiation . Overall , these observations reveal the clinical relevance and prognostic value of tumor-derived exosomes and identify a unique intercellular mechanism mediated by tumor-derived exosomes to modulate T-cell function in NPC . A novel approach in the treatment of cancer : targeting the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) autocrine pathway contributes to a number of processes important to cancer development and progression , including cell proliferation , apoptosis , angiogenesis , and metastatic spread . The critical role the P00533 plays in cancer has led to an extensive search for selective inhibitors of the P00533 signaling pathway . The results of a large body of preclinical studies and the early clinical trials thus far conducted suggest that targeting the P00533 could represent a significant contribution to cancer therapy . A variety of different approaches are currently being used to target the P00533 . The most promising strategies in clinical development include monoclonal antibodies to prevent ligand binding and small molecule inhibitors of the tyrosine kinase enzymatic activity to inhibit autophosphorylation and downstream intracellular signaling . At least five blocking monoclonal antibodies have been developed against the P00533 . Among these , IMC-225 is a chimeric human-mouse monoclonal IgG1 antibody that has been the first anti- P00533 targeted therapy to enter clinical evaluation in cancer patients in Phase II and III studies , alone or in combination with conventional therapies , such as radiotherapy and chemotherapy . A number of small molecule inhibitors of the P00533 tyrosine kinase enzymatic activity is also in development . DB00530 and ZD1839 ( DB00317 ) are currently in Phase II and III development , respectively . ZD1839 , a p.o. active , selective quinazoline derivative has demonstrated promising in vitro and in vivo antitumor activity . Preliminary results from Phase I and II trials in patients with advanced disease demonstrate that ZD1839 and DB00530 have an acceptable tolerability profile and promising clinical efficacy in patients with a variety of tumor types . This mini-review describes the P00533 inhibitors in clinical development . Interferon-alpha promotes the anti-proliferative effect of Erlotinib ( DB00530 ) on human colon cancer cell lines . Interferon-alpha ( IFNalpha ) treatment is associated with up-regulation of epidermal growth factor receptor ( P00533 / P00533 ) expression and marked growth inhibition while maintaining the sensitivity of the target colon cancer cells to epidermal growth factor ( Gut 2004;53:123 ) . We aimed to determine the effect of combining IFNalpha and Erlotinib ( an P00533 / P00533 inhibitor ) on colon cancer cell line growth . Crystal-violet staining and flow cytometry were used to assess cell proliferation and expression of P00533 / P00533 . IFNalpha pre-treatment followed by a combination of IFNalpha plus Erlotinib significantly enhanced the sensitivity of 7/9 of colon cancer cell lines by 7-43 % . This approach may have clinical implications for improving treatment based on targeting of P00533 / P00533 . Why the epidermal growth factor receptor ? The rationale for cancer therapy . There is a need for new , selective anticancer agents that differentiate between malignant and nonmalignant cells . The benefits of such agents would include a higher therapeutic index and lower toxicity than conventional therapies . Although expressed in nonmalignant cells , the epidermal growth factor receptor ( P00533 ) is highly expressed in a variety of tumors , and its expression correlates with poor response to treatment , disease progression , and poor survival . Evidence for a role for the P00533 in the inhibition and pathogenesis of various cancers has led to the rational design and development of agents that selectively target this receptor . Activation of the P00533 signaling pathway in cancer cells has been linked with increased cell proliferation , angiogenesis , and metastasis , and decreased apoptosis . Preclinical data show that anti- P00533 therapies can inhibit these effects in vitro and in vivo . In addition , preclinical data confirm that many such agents have the potential to increase the effectiveness of current cytotoxic agents . Following accelerated drug development programs , phase III trials are now under way for a number of P00533 -targeted therapies , including the monoclonal antibody IMC-C225 and the P00533 -tyrosine kinase inhibitors ZD1839 ( DB00317 ) and DB00530 . Thus , the rationale for P00533 -targeted approaches to cancer treatment is apparent and now well established , and there is increasing evidence that they may represent a significant contribution to cancer therapy . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Targeting epidermal growth factor receptor in lung cancer . Among the most promising agents in clinical development to treat non-small-cell lung cancer ( NSCLC ) are the epidermal growth factor receptor ( P00533 ) targeting agents . A series of recent studies have demonstrated the activity of anti- P00533 targeted therapies for NSCLC . In advanced NSCLC that is refractory to chemotherapy , antitumor responses have been reported with P00533 tyrosine kinase inhibitors ( ZD1839 and DB00530 ) . The role of ZD1839 and DB00530 as possible additions to standard chemotherapy in the first-line setting has also been evaluated , and the studies conducted to date should respond to the question of whether these compounds could provide a survival benefit . Other areas of research involve looking at the role of P00533 tyrosine kinase inhibitors in the neoadjuvant treatment of stage III NSCLC and the planning of chemoprevention studies . These exciting results and plans are further complemented by an emerging number of compounds in clinical development , including both monoclonal antibodies ( ie , IMC-C225 ) and other tyrosine kinase inhibitors , directed at the P00533 . New targeted therapies in gastrointestinal cancers . Despite surgical , radiotherapeutic , and chemotherapeutic advances , a large proportion of gastrointestinal ( GI ) cancers remain incurable . An improved understanding of the molecular pathogenesis of cancer has promulgated the development of novel agents designed to target critical pathways involved in cancer development and progression . The crucial role of the epidermal growth factor receptor ( P00533 ) in tumor proliferation and the overexpression of P00533 in several GI cancers provides the rationale for targeting and interrupting this key signaling network . P00533 blockade through monoclonal antibodies ( C225 and DB01269 ) and tyrosine kinase inhibitors ( ZD1839 and DB00530 ) has translated into promising evidence of clinical benefit . Ras-mediated signal transduction has been targeted using inhibitors of farnesyl transferase ( R115777 and SCH66336 ) to block the post-translation modification of Ras . Inhibitors of vascular growth factor receptor ( bevacizumab and PTK787 ) and matrix metalloproteinase target the effects of the host environment . P35354 inhibitors in colorectal cancer and STI571 in GI stromal tumors represent novel therapies of interest for these specific GI cancers . Evidence suggests that novel agents can be administered alone or in combination with standard therapies with little additional toxicity . The results of ongoing and future research efforts will clarify the optimal use and survival benefit of targeted therapies for patients with GI malignancies . P05121 Regulates the Invasive Phenotype in Human Cutaneous Squamous Cell Carcinoma . The emergence of highly aggressive subtypes of human cutaneous squamous cell carcinoma ( SCC ) often reflects increased autocrine/paracrine TGF-beta synthesis and epidermal growth factor receptor ( P00533 ) amplification . Cooperative TGF-beta/ P00533 signaling promotes cell migration and induces expression of both proteases and protease inhibitors that regulate stromal remodeling resulting in the acquisition of an invasive phenotype . In one physiologically relevant model of human cutaneous SCC progression , TGF-beta1+ P01133 stimulation increases the production of several matrix metalloproteinases ( MMPs ) , among the most prominent of which is P09238 -an MMP known to be elevated in SCC in situ . Activation of stromal plasminogen appears to be critical in triggering downstream MMP activity . Paradoxically , P05121 , the major physiological inhibitor of plasmin generation , is also upregulated under these conditions and is an early event in progression of incipient epidermal SCC . One testable hypothesis proposes that TGF-beta1+ P01133 -dependent P09238 elevation directs focalized matrix remodeling events that promote epithelial cell plasticity and tissue invasion . Increased P05121 expression serves to temporally and spatially modulate plasmin-initiated pericellular proteolysis , further facilitating epithelial invasive potential . Defining the complex signaling and transcriptional mechanisms that maintain this delicate balance is critical to developing targeted therapeutics for the treatment of human cutaneous malignancies . A unique structure for epidermal growth factor receptor bound to GW572016 ( DB01259 ) : relationships among protein conformation , inhibitor off-rate , and receptor activity in tumor cells . GW572016 ( DB01259 ) is a tyrosine kinase inhibitor in clinical development for cancer that is a potent dual inhibitor of epidermal growth factor receptor ( P00533 , ErbB-1 ) and ErbB-2 . We determined the crystal structure of P00533 bound to GW572016 . The compound is bound to an inactive-like conformation of P00533 that is very different from the active-like structure bound by the selective P00533 inhibitor DB00530 ( Tarceva ) described previously . Surprisingly , we found that GW572016 has a very slow off-rate from the purified intracellular domains of P00533 and ErbB-2 compared with DB00530 and another P00533 selective inhibitor , ZD-1839 ( DB00317 ) . Treatment of tumor cells with these inhibitors results in down-regulation of receptor tyrosine phosphorylation . We evaluated the duration of the drug effect after washing away free compound and found that the rate of recovery of receptor phosphorylation in the tumor cells reflected the inhibitor off-rate from the purified intracellular domain . The slow off-rate of GW572016 correlates with a prolonged down-regulation of receptor tyrosine phosphorylation in tumor cells . The differences in the off-rates of these drugs and the ability of GW572016 to inhibit ErbB-2 can be explained by the enzyme-inhibitor structures . Anti-epidermal growth factor receptor drugs in cancer therapy . The epidermal growth factor receptor is a cell membrane growth factor receptor that plays a key role in cancer development and progression . P00533 -activated signalling pathways control cell proliferation , apoptosis , angiogenesis and metastatic spread in the majority of human epithelial cancers . Targeting the epidermal growth factor receptor represents a valuable molecular approach to cancer therapy . Promising strategies in clinical development include monoclonal antibodies which block ligand binding and small molecule inhibitors of the tyrosine kinase enzymatic activity which prevent epidermal growth factor receptor autophosphorylation and propagation of downstream intracellular signals . Several anti-epidermal growth factor receptor agents are in clinical development for cancer therapy . Among these , IMC-225 ( cetuximab ) , a chimeric human-mouse monoclonal IgG1 antibody , DB00530 ( Tarceva ) and ZD1839 ( DB00317 ) , two small molecule epidermal growth factor receptor-selective tyrosine kinase inhibitors , are currently in Phase II and III development as single agents or in combination with conventional therapies , such as radiotherapy or chemotherapy . Results from Phase I - II trials in advanced cancer demonstrate that these drugs have an acceptable tolerability and an interesting clinical activity in patients with a variety of tumour types . Efficacy and safety of erlotinib HCl , an epidermal growth factor receptor ( P00533 / P00533 ) tyrosine kinase inhibitor , in patients with advanced ovarian carcinoma : results from a phase II multicenter study . The aim of this single-arm , phase II study was to estimate the tumor response rate and safety profile of erlotinib HCl ( erlotinib , Tarceva , DB00530 ) monotherapy in patients with refractory , recurrent , P00533 / P00533 -positive epithelial ovarian tumors , who had failed prior taxane and/or platinum-based chemotherapy . Thirty-four patients received 150 mg erlotinib orally once daily for up to 48 weeks or until disease progression or dose-limiting toxicity . Two patients had partial responses , lasting 8+ and 17 weeks , giving an objective response rate of 6 % ( 95 % confidence interval [ CI ] , 0.7-19.7 % ) . Fifteen patients ( 44 % ) had stable disease , and 17 patients ( 50 % ) had progressive disease . Median overall survival was 8 months ( 95 % CI , 5.7-12.7 months ) , with a 1-year survival rate of 35.3 % ( 95 % CI , 19.8-53.5 % ) . Patients with rash survived significantly longer than those without ( P= 0.009 ) , correlating with rash grade . Erlotinib was generally well tolerated . The most frequent erlotinib-related adverse events were rash ( 68 % ) and diarrhea ( 38 % ) . Erlotinib had marginal activity but was generally well tolerated . The safety profile appears more favorable than typically experienced with standard chemotherapeutic agents , which is encouraging in these heavily pretreated patients . Combination of erlotinib with chemotherapy or other targeted agents should be considered . Gefitinib ( ' DB00317 ' , ZD1839 ) and new epidermal growth factor receptor inhibitors . The epidermal growth factor receptor ( P00533 ) is a promising target for cancer therapy and a number of P00533 -targeted agents have been developed . Those most advanced in development are the P00533 tyrosine kinase inhibitors gefitinib ( ' DB00317 ' , ZD1839 ) and erlotinib ( ' Tarceva ' , DB00530 ) , and the monoclonal antibody cetuximab ( ' Erbitux ' , IMC-C225 ) . This review provides a clinical overview of these agents , highlighting their antitumour activities in different tumour types . P00533 -targeted agents are generally well tolerated and are not typically associated with the severe adverse events often seen with cytotoxic chemotherapy . Gefitinib is the agent with the most extensive clinical experience , particularly in non-small-cell lung cancer ( NSCLC ) . Recently , gefitinib became the first-approved P00533 -targeted agent , for use in patients with previously treated advanced NSCLC in Japan , the USA and other countries . Further studies are required to explore the full potential of these novel agents either as monotherapy or combination therapy . Erlotinib : success of a molecularly targeted agent for the treatment of advanced pancreatic cancer . P00533 ( P00533 ) is overexpressed by several solid tumors , including pancreatic cancer , and has become an important target for novel anticancer pharmacotherapy . Erlotinib ( Tarceva , DB00530 ) is an orally available small-molecule inhibitor of the P00533 tyrosine kinase . The addition of erlotinib to gemcitabine has been shown to prolong survival of patients treated for advanced pancreatic cancer in the National Cancer Institute of Canada PA.3 trial . This survival advantage is small yet noteworthy , in that numerous gemcitabine-containing combinations have failed to show a statistically significant survival advantage over gemcitabine alone . The most frequent toxicities associated with the addition of erlotinib are diarrhea and rash . Erlotinib-induced rash appears to be predictive of outcome . Further clinical studies of erlotinib in the treatment of pancreatic cancer are ongoing . Phase II clinical trial data with the epidermal growth factor receptor tyrosine kinase inhibitor erlotinib ( DB00530 ) in non-small-cell lung cancer . Erlotinib ( DB00530 ; Tarceva ) is an orally available , highly specific epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor . The results of 3 phase II studies with erlotinib in non-small-cell lung cancer ( NSCLC ) are reviewed herein : ( 1 ) in patients with chemotherapy-resistant , P00533 / P00533 -expressing NSCLC of all histologies , ( 2 ) in patients with bronchoalveolar carcinoma previously untreated or treated with chemotherapy , and ( 3 ) as first-line therapy in elderly patients with NSCLC of all histologies . These studies have evaluated tumor response , survival , and symptom improvement . Erlotinib was given as an oral , continuous daily dose of 150 mg . The drug was well tolerated ; drug-related cutaneous rash and diarrhea were observed in approximately two thirds of patients . Withdrawals caused by toxicity were rare . The response rates were 12.3 % , 25 % , and 13.3 % , respectively . Mature survival data are available for the first trial . The median survival was 8.4 months , and the 1-year survival rate was 40 % . All responding patients in the first and second trials presented skin rash . In addition , survival correlated with the occurrence and severity of rash in the first trial . No data on the correlation between rash and survival are available for the second and third trials . Erlotinib is active and well tolerated in patients with NSCLC as first- and second-line therapy . Cutaneous rash appears to be a surrogate marker of clinical benefit , but this finding needs to be confirmed in ongoing and future studies . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . DB00530 OSI Pharmaceuticals . DB00530 ( formerly CP-358774 ) , a quinazoline derivative , is an orally active epidermal growth factor receptor ( P00533 ) inhibitor which was originally under joint development by Pfizer and OSI Pharmaceuticals ( formerly Oncogene Science ) for the potential treatment of cancer ( eg , ovarian , non-small cell lung cancer ( NSCLC ) and head and neck ) . It is being evaluated in phase II trials [ 304305 ] , [ 372201 ] . On 8 January 2001 , OSI announced that it had signed an agreement with Roche and Genentech for the global co-development and marketing of DB00530 . The agreement with Genentech covers the United States , that with Roche the rest of the world [ 395371 ] , [ 395526 ] . In June 2000 , OSI gained all development and marketing rights for DB00530 following Pfizer 's merger with Warner-Lambert [ 371439 ] . In September 2000 , Pfizer transferred the IND dossierfor DB00530 to OSI ahead of the timeline agreed in the June 2000 development and marketing rights agreement [ 383786 ] . The phase II trials will assess DB00530 both as a single agent and in combination with existing chemotherapy regimens [ 347783 ] . Phase III trials are expected to be initiated in 2001 [ 347783 ] . In October 2000 , Lehman Brothers predicted that DB00530 would move into pivotal trials in thefirst half of 2001 and that the drug would be launched in 2003 . The analysts also estimated worldwide sales of US $ 66 million , $ 285 million and $ 461 million in 2003 , 2004 and 2005 , respectively , and peak sales in excess of US $ 500 million [ 395189 ] . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Structure of the epidermal growth factor receptor kinase domain alone and in complex with a 4-anilinoquinazoline inhibitor . The crystal structure of the kinase domain from the epidermal growth factor receptor ( EGFRK ) including forty amino acids from the carboxyl-terminal tail has been determined to 2.6-A resolution , both with and without an EGFRK-specific inhibitor currently in Phase III clinical trials as an anti-cancer agent , erlotinib ( DB00530 , CP-358,774 , Tarceva(TM) ) . The P00533 family members are distinguished from all other known receptor tyrosine kinases in possessing constitutive kinase activity without a phosphorylation event within their kinase domains . Despite its lack of phosphorylation , we find that the EGFRK activation loop adopts a conformation similar to that of the phosphorylated active form of the kinase domain from the insulin receptor . Surprisingly , key residues of a putative dimerization motif lying between the EGFRK domain and carboxyl-terminal substrate docking sites are found in close contact with the kinase domain . Significant intermolecular contacts involving the carboxyl-terminal tail are discussed with respect to receptor oligomerization . Development of the epidermal growth factor receptor inhibitor Tarceva ( DB00530 ) . The epidermal growth factor receptor ( P00533 ) is a transmembrane receptor involved in the regulation of a complex array of essential biological processes such as cell proliferation and survival . Dysregulation of P00533 signaling network has been frequently reported in multiple human cancers and has been associated with the processes of tumor development , growth , proliferation , metastasis and angiogenesis . Inhibition of the P00533 was associated with antitumor effects in preclinical models . On the bases of these data , therapeutics targeting the P00533 were explore in clinical trials . Tarceva ( DB00530 , OSI Pharmaceuticals , Uniondale , NY ) is a small molecule selective inhibitor of the P00533 tyrosine kinase ( TK ) . In preclinical studies , Tarceva inhibited the phosphorylation of the P00533 in a dose and concentration dependent manner resulting in cell cycle arrest and induction of apoptosis . In in vivo studies , the agent caused tumor growth inhibition and shoved synergistic effects when combined with conventional chemotherapy . Subsequent single agent phase I studies and phase I studies in combination with chemotherapy demonstrated that the agent has a good safety profile and induced tumor growth inhibition in a substantial number of patients with a variety of different solid tumor . Preliminary report from phase II studies confirmed the excellent tolerability of Tarceva as well as showed encouraging preliminary activity . Phase III studies have either been completed or are ongoing in several tumor types such as lung cancer and pancreatic cancer . In summary , Tarceva is a novel inhibitor of the P00533 TK which has shown promising activity in initial studies and is currently undergoing full development as an anticancer drug . Targeted therapy using novel agents in the treatment of non-small-cell lung cancer . Patients with advanced non-small-cell lung cancer ( NSCLC ) have a poor prognosis and high mortality . The therapeutic improvement caused by the new generation of cytotoxic agents seems to have reached a plateau . The main categories of targeted therapeutics applicable for NSCLC include receptor-targeted therapy , signal transduction or cell-cycle inhibition , angiogenesis inhibitors , gene therapy , and vaccines . Several major classes of agents directed at specific cellular mechanisms exist for the treatment of NSCLC . The anti-epidermal growth factor receptor ( P00533 ) group contains trastuzumab and IMC-C225 , monoclonal antibodies against EGFRs that are overexpressed in many cancers . DB00530 and ZD1839 are inhibitors of P00533 tyrosine kinase , a key enzyme of the signaling pathway . Farnesyl transferase inhibitors , such as SCH66336 , and protein kinase C inhibitors , such as ISIS 3521 , have also shown antitumor activity . Antiangiogenesis agents that have shown promise include TNP-470 , recombinant endostatin , and angiostatin . Antibodies to vascular endothelial growth factor ( P15692 ) also seem to control tumor progression and may prolong survival . LY317615 , an inhibitor of protein kinase Cb , augmented the tumor growth delay produced by cytotoxic drugs . All of these agents are in different phases of clinical testing and have shown encouraging activity as single agents or in combination with chemotherapy drugs . These new agents are more target specific , less toxic , easier to administer , and may lead to enhanced safety and survival for patients with advanced NSCLC . Biologically targeted treatment of non-small-cell lung cancer : focus on epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) has emerged in recent years as a key target of molecular therapy for solid tumors . The postembryonic role of P00533 is normally limited . In cancer , however , abnormal P00533 -tyrosine kinase ( TK ) activity plays a central role in many of the processes involved in tumor progression , such as proliferation , angiogenesis , invasiveness , decreased apoptosis , and loss of differentiation . Several different approaches have been taken to inhibit P00533 -mediated activity in tumor cells , including monoclonal antibodies directed at the ligand-binding portion of the P00533 and small-molecule agents that directly inhibit the intracellular TK domain of P00533 . Two of these TK inhibitors , gefitinib and erlotinib ( DB00530 , Tarceva ) , have shown antitumor activity and good tolerability across several tumor types in early dose-finding clinical trials , particularly for non-small-cell lung cancer ( NSCLC ) . In heavily pretreated patients with advanced NSCLC , gefitinib showed clinically significant tumor responses and symptom relief with good tolerability . Based on these results , gefitinib has now been approved for the third-line treatment of advanced NSCLC . The use of gefitinib in standard treatment programs or combined with other molecular targeted agents may substantially improve the outlook for patients with NSCLC or other types of solid tumors Significance of interleukin-6 signaling in the resistance of pharyngeal cancer to irradiation and the epidermal growth factor receptor inhibitor . PURPOSE : Tumor eradication by chemoradiotherapy for pharyngeal cancer has not been particularly successful . Targeting epithelial growth factor receptor ( P00533 ) could be a potential treatment strategy providing additional benefits , but only a subset of these tumors gives a clinically significant response to P00533 inhibitors . The aim has been to identify the role of interleukin-6 ( P05231 ) signaling and its predictive power in the treatment response of pharyngeal cancer . METHODS AND MATERIALS : Human pharyngeal cancer cell lines , including the hypopharyngeal cancer cell line FaDu and its derived cell line FaDu-C225-R , were selected . Changes in tumor growth , response to treatment , and responsible signaling pathway were investigated in vitro . Furthermore , 95 pharyngeal cancer tissue specimens were analyzed by immunohistochemical staining , and correlations were made between levels of P05231 , P05231 receptor ( IL-6R ) , p-AKT , and p- P40763 expression and the clinical outcome of patients . RESULTS : In vitro , either extrinsic P05231 stimulation of cancer cells or intrinsically activated P05231 signaling detected in FADu-C225-R cells results in resistance to irradiation and P00533 inhibitor . Blocking P05231 signaling attenuated aggressive tumor behavior and sensitized the cells to treatments . The responsible mechanisms included decreased p- P40763 , less nuclear translocation of P00533 , and subsequently attenuated epithelial-mesenchymal transition . Regarding clinical data , staining of p- P40763 and P05231 was significantly linked with lower response rates to treatments and shorter survival in pharyngeal cancer patients . CONCLUSIONS : P05231 and p- P40763 may be significant predictors of pharyngeal carcinoma , and regulating P05231 signaling can be considered a promising therapeutic approach . [ Molecular target-based cancer therapy : epidermal growth factor receptor inhibitors ] . Based on recent progress in cancer biology , numerous molecules that contribute to proliferation , invasion , and metastasis of cancer cells have been identified . The epidermal growth factor receptor ( P00533 ) , a member of cell membrane receptors , is overexpressed by many tumors , and P00533 overexpression correlates with poor prognosis and disease progression . The P00533 is an attractive target for novel anticancer therapy . ZD1839 and DB00530 , highly specific P00533 tyrosine kinase inhibitors , have shown promising antitumor activity against cisplatin-resistant non-small cell lung cancer in phase I and phase II trials . IMC-C225 , a monoclonal antibody against P00533 , has achieved significant disease control in head and neck cancer and colorectal cancer in combination with anticancer agents . These agents are under evaluation in phase III trials . In conclusion , it is expected that P00533 -directed therapies will soon be established as an effective novel treatment for many cancer patients . [ Cellular adhesion signal transduction network of tumor necrosis factor-alpha induced hepatocellular carcinoma cells ] . OBJECTIVE : To systemically explore the cellular adhesion signal transduction network of tumor necrosis factor-alpha ( P01375 -α ) -induced hepatocellular carcinoma cells with bioinformatics tools . METHODS : Published microarray dataset of P01375 -α-induced HepG2 , human transcription factor database HTRI and human protein-protein interaction database HPRD were used to construct and analyze the signal transduction network . RESULTS : In the signal transduction network , MYC and SP1 were the key nodes of signaling transduction . Several genes from the network were closely related with cellular adhesion. P00533 ( P00533 ) is a possible key gene of effectively regulating cellular adhesion during the induction of P01375 -α . CONCLUSION : P00533 is a possible key gene for P01375 -α-induced metastasis of hepatocellular carcinoma .

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interacts_with DB01241?

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LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Regulation of angiotensin II receptors beyond the classical pathway . The DB01367 ( renin-angiotensin system ) plays a role not only in the cardiovascular system , including blood pressure regulation , but also in the central nervous system . AngII ( angiotensin II ) binds two major receptors : the AT(1) receptor ( AngII type 1 receptor ) and AT(2) receptor ( AngII type 2 receptor ) . It has been recognized that AT(2) receptor activation not only opposes AT(1) receptor actions , but also has unique effects beyond inhibitory cross-talk with AT(1) receptor signalling . Novel pathways beyond the classical actions of DB01367 , the P12821 ( angiotensin-converting enzyme ) /AngII/AT(1) receptor axis , have been highlighted : the Q9BYF1 /Ang-(1-7) [ angiotensin-(1-7) ] /Mas receptor axis as a new opposing axis against the P12821 /AngII/AT(1) receptor axis , novel AngII-receptor-interacting proteins and various AngII-receptor-activation mechanisms including dimer formation . Q6RW13 ( AT(1)-receptor-associated protein ) and Q9ULD2 ( AT(2)-receptor-interacting protein ) are well-characterized AngII-receptor-associated proteins . These proteins could regulate the functions of AngII receptors and thereby influence various pathophysiological states . Moreover , the possible cross-talk between Q07869 ( peroxisome-proliferator-activated receptor ) -γ and AngII receptor subtypes is an intriguing issue to be addressed in order to understand the roles of DB01367 in the metabolic syndrome , and interestingly some ARBs ( AT(1)-receptor blockers ) have been reported to have an AT(1)-receptor-blocking action with a partial Q07869 -γ agonistic effect . These emerging concepts concerning the regulation of AngII receptors are discussed in the present review . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . [ The effect of blood pressure-reducing therapy with captopril on tubular marker excretion in type-1 diabetics with nephropathy ] . A prospective open clinical trial was carried out with 23 hypertensive type I diabetics ( 13 men , ten women , mean age 49 +/- 9.1 years , duration of diabetes 18 +/- 9.1 years ) with early nephropathy . Glomerular and tubular renal function and metabolic parameters were monitored during 8 months ' treatment with the angiotensin converting enzyme ( P12821 ) inhibitor , captopril , in addition to previous antihypertensive treatment with one or more drugs . Blood pressure control tended to improve on captopril ( systolic pressures 152 +/- 13 vs 140 +/- 13 mm Hg , P < 0.05 ; diastolic pressures 89 +/- 10 vs 87 +/- 10 mm Hg , not significant ) . Proteinuria ( > 0.5 g/24 hours ) fell into the microalbuminuria range ( albumin excretion 2-20 mg/mmol creatinine ) in four out of 13 patients , and microalbuminuria disappeared in four out of ten patients . Urinary levels of the brush border enzyme O60502 ( NAG ) , a marker of tubular dysfunction , were initially raised and fell significantly after 8 months ' treatment with captopril ( 20.3 +/- 14.4 vs 8.8 +/- 8.1 U/g creatinine ; P < 0.01 ) . DB01197 did not affect metabolic control ( HbA1 , total , HDL and LDL cholesterol , triglycerides , apolipoproteins A1 and B ) or the insulin dosage . These results show that long-term treatment with captopril may favourably influence both albumin excretion and NAG activity , a marker of tubular dysfunction , in type I diabetics with nephropathy . 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 . Peroxisome proliferator-activated receptor-gamma is a target of nonsteroidal anti-inflammatory drugs mediating cyclooxygenase-independent inhibition of lung cancer cell growth . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) inhibit the growth of different cancer cell types , suggesting a broad role for their cyclooxygenase ( P36551 ) targets and eicosanoid products in tumor cell growth . Sulindac sulfide , a P36551 inhibitor , inhibited the growth of non-small-cell lung cancers ( NSCLC ) both in soft agar and as xenografts in nude mice . Importantly , the concentration of sulindac sulfide required to inhibit NSCLC cell growth greatly exceeded the concentration required to inhibit prostaglandin ( PG ) E(2) synthesis in NSCLC cells , suggesting that NSAID inhibition of cell growth is mediated by additional targets distinct from P36551 . Both sulindac sulfide and ciglitazone , a defined peroxisome proliferator-activated receptor-gamma ( PPARgamma ) agonist , stimulated a promoter construct containing a Q07869 response element linked to luciferase and potently inhibited NSCLC cell growth at similar concentrations , indicating a role for PPARgamma as a target of NSAID action in these cells . Overexpression of PPARgamma in NSCLC cells strongly inhibited the transformed growth properties of the cells , providing a molecular confirmation of the results obtained with the PPARgamma agonists . Increased expression of PPARgamma , as well as ciglitazone and sulindac sulfide induced expression of P12830 , which has been linked to increased differentiation of NSCLC . Despite the fact that SCLC cell lines expressed little or no cytosolic phospholipase A(2) , P23219 , or P35354 , sulindac sulfide and PPARgamma agonists also inhibited the transformed growth of these lung cancer cells . We propose that PPARgamma serves as a target for NSAIDs that accounts for P36551 -independent inhibition of lung cancer cell growth . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . P60568 suppression by 2-arachidonyl glycerol is mediated through peroxisome proliferator-activated receptor gamma independently of cannabinoid receptors 1 and 2 . 2-Arachidonyl glycerol ( 2-AG ) is an endogenous arachidonic acid derivative that binds cannabinoid receptors P21554 and CB2 and is hence termed an endocannabinoid . 2-AG also modulates a variety of immunological responses , including expression of the autocrine/paracrine T cell growth factor interleukin ( IL ) -2 . The objective of the present studies was to determine the mechanism responsible for P60568 suppression by 2-AG . Because of the labile properties of 2-AG , 2-AG ether , a nonhydrolyzable analog of 2-AG , was also used . Both 2-AG and 2-AG ether suppressed P60568 expression independently of P21554 and CB2 , as demonstrated in leukocytes derived from P21554 /CB2-null mice . Moreover , we demonstrated that both 2-AG and 2-AG ether treatment activated peroxisome proliferator-activated receptor gamma ( PPARgamma ) , as evidenced by forced differentiation of 3T3- Q9NUQ9 cells into adipocytes , induction of aP2 mRNA levels , and activation of a PPARgamma-specific luciferase reporter in transiently transfected 3T3- Q9NUQ9 cells . Consequently , the putative role of PPARgamma in P60568 suppression by 2-AG and 2-AG ether was examined in Jurkat T cells . Concordant with PPARgamma involvement , the PPARgamma-specific antagonist 2-chloro-5-nitro-N-(4-pyridyl)-benzamide ( T0070907 ) blocked 2-AG- and 2-AG ether-mediated P60568 suppression . Likewise , 2-AG suppressed the transcriptional activity of two transcription factors crucial for P60568 expression , nuclear factor of activated T cells and nuclear factor kappaB , in the absence but not in the presence of T0070907 . 2-AG treatment also induced PPARgamma binding to a Q07869 response element in activated Jurkat T cells . Together , the aforementioned studies identify PPARgamma as a novel intracellular target of 2-AG , which mediates the suppression of P60568 by 2-AG in a manner that is independent of P21554 and/or CB2 . Involvement of PPARs in Cell Proliferation and Apoptosis in Human Colon Cancer Specimens and in Normal and Cancer Cell Lines . Q07869 involvement in cell growth was investigated " in vivo " and " in vitro " and was correlated with cell proliferation and apoptotic death . " In vivo " PPARgamma and alpha were evaluated in colon cancer specimens and adjacent nonneoplastic colonic mucosa . PPARgamma increased in most cancer specimens versus mucosa , with a decrease in c-Myc and in P12004 proteins , suggesting that colon cancer growth is due to increased cell survival rather than increased proliferation . The prevalence of survival over proliferation was confirmed by Bcl-2 or Bcl-X(L) increase in cancer versus mucosa , and by decreased PPARalpha . " In vitro " PPARgamma and PPARalpha were evaluated in human tumor and normal cell lines , treated with natural or synthetic ligands . PPARgamma was involved in inhibiting cell proliferation with a decrease in c-Myc protein , whereas PPARalpha was involved in inducing apoptosis with modulation of Bcl-2 and Bad proteins . This involvement was confirmed using specific antagonists of two PPARs . Moreover , the results obtained on treating cell lines with Q07869 ligands confirm observations in colon cancer : there is an inverse correlation between PPARalpha and Bcl-2 and between PPARgamma and c-Myc . [ The interconnections of molecular mechanisms of hormone actions and their role in pathogenesis of obesity , insulin resistance , and diabetes mellitus ] . The various hormones , proteins and other compounds related to developing obesity , insulin resistance and type 2 diabetes are analyzed in the paper . 1 ) Leptin , ciliary neurutrophic factor , adiponectin , glucagon-like peptide 1 , peptide YY , neuromedin S , as well as the protein receptors of these hormones decrease the food consumption , increase the energy turnover , and prevent obesity , insulin resistance , and type 2 diabetes development . The mediators of these hormone and receptor actions are melanocyte stimulating hormone ( MSH ) , corticotropin-releasing hormone ( P06850 ) , and the others . 2 ) Ghrelin , endogenose cannabinoides , galanin-like peptide and the mediators of their actions : neuropeptide Y ( P01303 ) and Agouti gene related protein ( O00253 ) increase the appetite and food consumption . Peroxisome proliferation-activated receptor ( Q07869 ) performs the similar action on food intake . The activation of the first group compound functioning decreases the obesity , increases the energy turnover , facilitates the insulin action and prevents the insulin resistance and type 2 diabetes . Increasing the activities of the second group , as well as , decreasing the actions of the first one of substances induce the opposite effects and facilitate obesity , insulin resistance , and type 2 diabetes developments . The interconnections of the molecular mechanisms of so many hormone actions make the very complicated tusk to study the various endocrine disorders including diabetes mellitus as well . Molecular targets and regulators of cardiac hypertrophy . Cardiac hypertrophy is one of the main ways in which cardiomyocytes respond to mechanical and neurohormonal stimuli . It enables myocytes to increase their work output , which improves cardiac pump function . Although cardiac hypertrophy may initially represent an adaptive response of the myocardium , ultimately , it often progresses to ventricular dilatation and heart failure which is one of the leading causes of mortality in the western world . A number of signaling modulators that influence gene expression , apoptosis , cytokine release and growth factor signaling , etc. are known to regulate heart . By using genetic and cellular models of cardiac hypertrophy it has been proved that pathological hypertrophy can be prevented or reversed . This finding has promoted an enormous drive to identify novel and specific regulators of hypertrophy . In this review , we have discussed the various molecular signal transduction pathways and the regulators of hypertrophic response which includes calcineurin , cGMP , NFAT , natriuretic peptides , histone deacetylase , P05231 cytokine family , Gq/ P49842 signaling , PI3K , MAPK pathways , Na/H exchanger , DB01367 , polypeptide growth factors , P01160 , NO , P01375 , Q07869 and JAK/ P35610 pathway , microRNA , Cardiac angiogenesis and gene mutations in adult heart . Augmented knowledge of these signaling pathways and their interactions may potentially be translated into pharmacological therapies for the treatment of various cardiac diseases that are adversely affected by hypertrophy . The purpose of this review is to provide the current knowledge about the molecular pathogenesis of cardiac hypertrophy , with special emphasis on novel researches and investigations . 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 . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . Obesity and breast cancer : the roles of peroxisome proliferator-activated receptor-γ and plasminogen activator inhibitor-1 . Breast cancer is the most prominent cancer among females in the United States . There are a number of risk factors associated with development of breast cancer , including consumption of a high-fat diet and obesity . P00747 activator inhibitor-1 ( P05121 ) is a cytokine upregulated in obesity whose expression is correlated with a poor prognosis in breast cancer . As a key mediator of adipogenesis and regulator of adipokine production , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) is involved in P05121 expression from adipose tissue . We summarize the current knowledge linking Q07869 -γ and P05121 expression to high-fat diet and obesity in the risk of breast cancer . Q07869 gene variants influence progression of coronary atherosclerosis and risk of coronary artery disease . BACKGROUND : Q07869 ( PPARalpha ) regulates the expression of genes involved in lipid metabolism and inflammation , making it a candidate gene for atherosclerosis and ischemic heart disease ( IHD ) . METHODS AND RESULTS : We investigated the association between the leucine 162 to valine ( L162V ) polymorphism and a G to C transversion in intron 7 of the PPARalpha gene and progression of atherosclerosis in the DB01241 Coronary Angiography Trial ( LOCAT ) , a trial examining the effect of gemfibrozil treatment on progression of atherosclerosis after bypass surgery and on risk of IHD in the second Northwick Park Heart Study ( Q9NP85 ) , a prospective study of healthy middle-aged men in the United Kingdom . There was no association with plasma lipid concentrations in either study . Both polymorphisms influenced progression of atherosclerosis and risk of IHD . V162 allele carriers had less progression of diffuse atherosclerosis than did L162 allele homozygotes with a similar trend for focal atherosclerosis . Intron 7 C allele carriers had greater progression of atherosclerosis than did G allele homozygotes . The V162 allele attenuated the proatherosclerotic effect of the intron 7 C allele . Homozygotes for the intron 7 C allele had increased risk of IHD , an effect modulated by the L162V polymorphism CONCLUSIONS : The PPARalpha gene affects progression of atherosclerosis and risk of IHD . Absence of association with plasma lipid concentrations suggests that PPARalpha affects atherosclerotic progression directly in the vessel wall . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples .

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Effects of inhaled prostacyclin analogue on chronic hypoxic pulmonary hypertension . Inhaled DB01240 has been reported to elicit pulmonary vasodilation , but whether it is also effective in treating chronic hypoxic pulmonary hypertension is still uncertain . We designed this study to address the in vivo effectiveness of inhaled DB05229 , a stable DB01240 analogue , on pulmonary vascular tone during hypoxic exposure in normoxic ( N ) and chronically hypoxic ( CH ) rats . Pulmonary vasodilation was observed by low-dose inhaled DB05229 in N rats , but not in CH rats . It was not until higher doses of DB05229 were given that pulmonary vasodilation was obtained in CH rats . When the agent was continuously administered by an intravascular route at the inhaled dose , it elicited no vasodilation in N rats . On the contrary , it elicited profound vasodilation in CH rats , although a concomitant systemic hypotension was observed . The P43119 mRNA expression was unchanged in the lungs of CH rats compared with that of N rats . We conclude that low doses of aerosolized DB05229 may reduce pulmonary vascular tone in rats without preexisting lung diseases . In contrast , when hypoxic pulmonary hypertension is present , the threshold of DB05229 inhalation was elevated to provoke pulmonary vasodilation . Population pharmacokinetic study of memantine : effects of clinical and genetic factors . BACKGROUND AND OBJECTIVE : Memantine , a frequently prescribed anti-dementia drug , is mainly eliminated unchanged by the kidneys , partly via tubular secretion . Considerable inter-individual variability in plasma concentrations has been reported . We aimed to investigate clinical and genetic factors influencing memantine disposition . METHODS : A population pharmacokinetic study was performed including data from 108 patients recruited in a naturalistic setting . Patients were genotyped for common polymorphisms in renal cation transporters ( O15245 /2/5 , Q96FL8 , P08183 ) and nuclear receptors ( O75469 , Q14994 , RXR , Q07869 ) involved in transporter expression . RESULTS : The average clearance was 5.2 L/h with a 27 % inter-individual variability ( percentage coefficient of variation ) . Glomerular filtration rate ( p = 0.007 ) and sex ( p = 0.001 ) markedly influenced memantine clearance . O75469 rs1523130 was identified as the unique significant genetic covariate for memantine clearance ( p = 0.006 ) , with carriers of the O75469 rs1523130 CT/TT genotypes presenting a 16 % slower memantine elimination than carriers of the CC genotype . CONCLUSION : The better understanding of inter-individual variability of memantine disposition might be beneficial in the context of individual dose optimization . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Opposite effect of Hsp90α and Hsp90β on P29474 ability to produce nitric oxide or superoxide anion in human embryonic kidney cells . Heat shock protein 90 subfamily is composed by two cytosolic isoforms known as Hsp90α and Hsp90β . Endothelial nitric oxide synthase ( P29474 ) is regulated by Hsp90 , however the specific role of each Hsp90 isoform on NO production has not been established . This study was designed to evaluate the effect of Hsp90α and Hsp90β over-expression on P29474 /NO pathway . Rat Hsp90α and Hsp90β were cloned into pcDNA3.1(+) and transfected in human embryonic kidney cells ( P29320 -293 ) . Hsp90α and Hsp90β transfection was corroborated by Western blot analysis and their effect on NO production ( NO(2)/NO(3) ) , P29474 protein and its phosphorylation at Ser1177 and Thr495 , as well as Akt/ P31749 Ser473 phosphorylation was determined . The interaction of Hsp90α and Hsp90β with P29474 and the dimer/monomer ratio of Hsp90 , as well as O(2)(-) generation were also assessed . After transfection , Hsp90α and Hsp90β levels were significantly increased in P29320 -293 cells . The Hsp90α over-expression induced a significant increase in NO(2)/NO(3) levels , an effect that was associated with increased phosphorylation of P29474 DB00133 1177 and Akt/ P31749 Ser473 , as well as with a greater Hsp90α dimerization . Noteworthy , pcHsp90β transfection reduced significantly NO(2)/NO(3) and increased O(2)(-) generation . These effects were associated with a reduction of P29474 dimeric conformation , increased P29474 Thr495 phosphorylation , reduced Akt/ P31749 phosphorylation , and by a greater amount of monomeric Hsp90β conformation . These data show for first time that Hsp90α and Hsp90β differentially modulate NO and O(2)(-) generation by P29474 through promoting changes in P29474 conformation and phosphorylation state . Q9H4A3 activates large-conductance Ca2+-activated K+ channels through modulation of P27361 /2 signaling . With no lysine ( WNK ) kinases are members of the serine/threonine kinase family . We previously showed that Q96J92 inhibits renal large-conductance Ca(2+)-activated K(+) ( BK ) channel activity by enhancing its degradation through a lysosomal pathway . In this study , we investigated the effect of Q9H4A3 on Q12791 activity . In HEK293 cells stably expressing the α subunit of BK ( P29320 -BKα cells ) , siRNA-mediated knockdown of Q9H4A3 expression significantly inhibited both BKα channel activity and open probability . Knockdown of Q9H4A3 expression also significantly inhibited BKα protein expression and increased P27361 /2 phosphorylation , whereas overexpression of Q9H4A3 significantly enhanced BKα expression and decreased P27361 /2 phosphorylation in a dose-dependent manner in HEK293 cells . Knockdown of P27361 /2 prevented Q9H4A3 siRNA-mediated inhibition of BKα expression . Similarly , pretreatment of P29320 -BKα cells with the lysosomal inhibitor bafilomycin A1 reversed the inhibitory effects of Q9H4A3 siRNA on BKα expression in a dose-dependent manner . Knockdown of Q9H4A3 expression also increased the ubiquitination of BKα channels . Notably , mice fed a high-K(+) diet for 10 days had significantly higher renal protein expression levels of BKα and Q9H4A3 and lower levels of P27361 /2 phosphorylation compared with mice fed a normal-K(+) diet . These data suggest that Q9H4A3 enhances Q12791 function by reducing P27361 /2 signaling-mediated lysosomal degradation of the channel . P35354 -derived prostacyclin modulates vascular remodeling . Suppression of prostacyclin ( DB01240 ) biosynthesis may explain the increased incidence of myocardial infarction and stroke which has been observed in placebo controlled trials of cyclooxygenase ( P36551 ) -2 inhibitors . Herein , we examine if P35354 -derived DB01240 might condition the response of the vasculature to sustained physiologic stress in experimental models that retain endothelial integrity . Deletion of the P43119 ( IP ) or suppression of DB01240 with the selective P35354 inhibitor , nimesulide , both augment intimal hyperplasia while preserving luminal geometry in mouse models of transplant arteriosclerosis or flow-induced vascular remodeling . Moreover , nimesulide or IP deletion augments the reduction in blood flow caused by common carotid artery ligation in wild-type mice . Generation of both thromboxane (Tx)A2 and the isoprostane , 8 , 12 -iso iPF(2alpha)-VI , are increased in the setting of flow reduction and the latter increases further on administration of nimesulide . Deletion of the TxA2 receptor ( TP ) reduces the hyperplastic response to nimesulide and carotid ligation , despite further augmentation of TP ligand production . Suppression of P35354 -derived DB01240 or deletion of IP profoundly influences the architectural response of the vasculature to hemodynamic stress . Mechanism based vascular remodeling may interact with a predisposition to hypertension and atherosclerosis in contributing to the gradual transformation of cardiovascular risk during extended periods of treatment with selective inhibitors of P35354 . Evidence for distinct prostaglandin I2 and D2 receptors in human platelets . Incubation of human platelet-rich plasma with prostaglandin I2 ( DB01240 ) , results in a marked increase in adenosine 3':5'-monophosphate ( DB02527 ) that persists for at least 60 min . The persistent stimulation of DB02527 levels by DB01240 can be rapidly reversed by the addition of either prostaglandin E1 or E2 ( PGE1 , DB00917 ) , but not by prostaglandin D2 ( PGD2 ) . Studies of agonist-specific desensitization of DB02527 accumulation show that PGE1 or DB00917 can desensitize for subsequent PGE or DB01240 activation , and that subthreshold levels of DB01240 desensitize for subsequent PGE1 stimulation . PGD2 desensitizes for consequent PGD2 activation , but not for PGE1 , DB00917 or DB01240 , and PGE compounds and DB01240 do not desensitize for subsequent PGD2 activation . Agonist-specific desensitization for DB01240 is not dependent on DB02527 accumulation , but appears to be a consequence of receptor occupation . Support of the desensitization experiments was obtained through the use of the prostaglandin antagonist N-0164 [ sodium-p-benzyl-4-[-oxo-2-(4-chlorobenzyl)-3-phenyl-propyl]phenyl phosphonate ) . This compound proved to be a potent antagonist of PGD2 and a weak antagonist of DB01240 -stimulated DB02527 accumulation . These data indicate that human platelets have distinct pharmacological receptors for both DB01240 and PGD2 , and that PGE compounds may actually interact with a P43119 . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Mediator subunit Gal11p/ Q96RN5 is required for fatty acid-dependent gene activation by yeast transcription factor Oaf1p . The yeast zinc cluster transcription factor Oaf1p activates transcription of target genes in response to direct binding of fatty acids in a manner analogous to the vertebrate nuclear receptor peroxisome proliferator-activated receptoralpha ( PPARalpha ) . PPARs and other metazoan nuclear receptors productively engage several distinct LXXLL motif-containing co-activators , including P52701 family members and the Q15648 /MED1 subunit of the Mediator co-activator , to promote ligand-dependent gene activation . Yeast , however , does not appear to harbor LXXLL motif co-activators , and the mechanism of fatty acid-dependent gene activation by the yeast PPARalpha analog Oaf1p is unknown . Here we show that the yeast Mediator subunit Gal11p/ Q96RN5 and its activator-targeted KIX domain plays a critical role in fatty acid-dependent transcriptional regulation of fatty acid beta-oxidation and peroxisomal genes by Oaf1p and for the ability of yeast to utilize fatty acids as a sole carbon source . Moreover , structural studies by NMR spectroscopy reveal that the Oaf1p activation domain interacts with the Gal11p/ Q96RN5 KIX domain in a manner similar to the yeast zinc cluster family member and xenobiotic receptor Pdr1p , revealing that the Gal11p/ Q96RN5 KIX domain is a key target of several ligand-dependent transcription factors in yeast . Together with previous work showing that the Caenorhabditis elegans Gal11p/ Q96RN5 homolog MDT-15 plays a critical role in regulation of fatty acid metabolism by the nematode Q07869 -like nuclear receptor NHR-49 , the findings presented here provide evidence for an ancient and essential role of a Mediator co-activator subunit in regulation of fatty acid metabolism by nuclear receptor-like transcription factors in eukaryotes . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . P43119 up-regulates the expression of angiogenic genes in human endometrium via cross talk with epidermal growth factor Receptor and the extracellular signaling receptor kinase 1/2 pathway . DB01240 ( P06744 ) is a member of the prostanoid family of lipid mediators that mediates its effects through a seven-transmembrane G protein-coupled receptor ( IP receptor ) . Recent studies have ascertained a role for prostanoid-receptor signaling in angiogenesis . In this study we examined the temporal-spatial expression of the IP receptor within normal human endometrium and additionally explored the signaling pathways mediating the role of IP receptor in activation of target angiogenic genes . Quantitative RT-PCR analysis demonstrated the highest endometrial expression of the IP receptor during the menstrual phase compared with all other stages of the menstrual cycle . Immunohistochemical analysis localized the site of IP receptor expression to the glandular epithelial compartment with stromal and perivascular cell immunoreactivity . Expression of the immunoreactive IP receptor protein was greatest during the proliferative and early secretory phases of the menstrual cycle . To explore the role of the IP receptor in glandular epithelial cells , we used the Ishikawa endometrial epithelial cell line . Stimulation of Ishikawa cells and human endometrial biopsy explants with 100 nm iloprost ( a P06744 analog ) rapidly activated P27361 /2 signaling and induced the expression of proangiogenic genes , basic fibroblast growth factor , angiopoietin-1 , and angiopoietin-2 , in an epidermal growth factor receptor ( P00533 ) -dependent manner . Furthermore , P00533 colocalized with IP receptor in the glandular epithelial compartment . These data suggest that P06744 -IP interaction within glandular epithelial cells can promote the expression of proangiogenic genes in human endometrium via cross talk with the P00533 . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . DB05229 sodium , a stable prostacyclin analogue , elicits dilation of isolated porcine retinal arterioles : roles of P29474 and potassium channels . PURPOSE : DB01240 ( DB01240 ) is usually described as an endoEDRFsthelium-derived relaxing factor , but the vasoreactivity to DB01240 in the retinal arterioles and the underlying mechanisms are not fully understood . We examined the effects of DB01240 on the retinal microcirculation using beraprost sodium ( BPS ) , a stable DB01240 analogue , and the signaling mechanisms involved in this vasomotor activity . METHODS : Porcine retinal arterioles were isolated , cannulated , and pressurized without flow in vitro . Video microscopic techniques recorded the diametric responses to BPS . RESULTS : DB05229 sodium elicited dose-dependent ( 0.1 pM-0.1 μM ) vasodilation of the retinal arterioles that was abolished by the P43119 ( IP ) antagonist CAY10441 . DB05229 sodium-induced vasodilation decreased by 50 % after the endothelium was removed and was inhibited by the nitric oxide ( NO ) synthase inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) comparable with denudation . Inhibition of soluble guanylyl cyclase by 1H-1,2,4-oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) and blockage of protein kinase A ( PKA ) by Rp-8-Br-cAMPS were comparable to L-NAME . DB05229 sodium-induced vasodilation was also inhibited by the nonselective potassium channel inhibitor , tetraethylammonium , and the adenosine triphosphate-sensitive potassium ( KATP ) channel blocker , glibenclamide . Residual vasodilation in the presence of glibenclamide decreased further with subsequent application of ODQ . CONCLUSIONS : DB05229 sodium , a stable DB01240 analogue , causes vasodilation of the retinal arterioles mediated via the IP receptor . The current findings suggest that BPS elicits endothelium-dependent and -independent dilation of the retinal arterioles mediated by NO induced by activation of PKA in the endothelium and the KATP channel activation in the vascular smooth muscle , respectively . Chromosomal localization of the human prostanoid receptor gene family . Prostaglandins ( PGD2 , DB00917 , PGF2 alpha , and DB01240 ) and thromboxane A2 ( TXA2 ) are biologically active molecules derived from the metabolism of arachidonic acid by cyclooxygenases . They produce a wide variety of physiological and pathophysiological effects mediated through specific G protein-coupled cell surface receptors . In this study , we have mapped the chromosomal positions of the human genes that encode the DB00917 receptor subtypes ( P34995 , PTGER2 , and P43115 ) , the PGF2 alpha receptor ( P43088 ) , the P43119 ( P43119 ) , and the TXA2 receptor ( P21731 ) using in situ hybridization . The P34995 , P21731 , and P43119 genes mapped to chromosome 19 at positions 19p13.1 , 19p13.3 , and 19q13.3 , respectively . The P43088 and P43115 genes mapped to chromosome 1 at positions 1p31.1 and 1p31.2 , respectively , and PTGER2 gene mapped to chromosome band 5p13.1 . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Role of prostacyclin in the cardiovascular response to thromboxane A2 . Thromboxane ( Tx ) A2 is a vasoconstrictor and platelet agonist . DB00945 affords cardioprotection through inhibition of TxA2 formation by platelet cyclooxygenase ( P23219 ) . DB01240 ( DB01240 ) is a vasodilator that inhibits platelet function . Here we show that injury-induced vascular proliferation and platelet activation are enhanced in mice that are genetically deficient in the P43119 ( IP ) but are depressed in mice genetically deficient in the TxA2 receptor ( TP ) or treated with a TP antagonist . The augmented response to vascular injury was abolished in mice deficient in both receptors . Thus , DB01240 modulates platelet-vascular interactions in vivo and specifically limits the response to TxA2 . This interplay may help explain the adverse cardiovascular effects associated with selective P35354 inhibitors , which , unlike aspirin and nonsteroidal anti-inflammatory drugs ( NSAIDs ) , inhibit DB01240 but not TxA2 . Nuclear factor kappa B inhibition improves conductance artery function in type 2 diabetic mice . BACKGROUND : We previously reported that enhanced nuclear factor kappa B ( NFκB ) activity is responsible for resistance arteries dysfunction in type 2 diabetic mice . METHODS : In this study , we aimed to determine whether augmented NFκB activity also impairs conductance artery ( thoracic aorta ) function in type 2 diabetic mice . We treated type 2 diabetic ( db(-) /db(-) ) and control ( db(-) /db(+) ) mice with two NFκB inhibitors ( dehydroxymethylepoxyquinomicin , 6 mg/kg , twice a week and IKK-NBD peptide , 500 µg/kg/day ) for 4 weeks . RESULTS : As expected , the NFκB inhibition did not affect blood glucose level and body weight . Thoracic aorta vascular endothelium-dependent relaxation ( EDR ) , determined by the wire myograph , was impaired in diabetic mice compared with control and was significantly improved after NFκB inhibition . Interestingly , thoracic EDR was also rescued in db(-) /db(-p50NFκB-/-) and db(-) /db(- P09874 -/-) double knockout mice compared with db(-) /db(-) mice . Similarly , the acute in vitro down regulation of NFκB-p65 using p65 shRNA lentiviral particles in arteries from db(-) /db(-) mice also improved thoracic aorta EDR . Western blot analysis showed that the p65NFκB phosphorylation , cleaved P09874 and P35354 expression were increased in thoracic aorta from diabetic mice , which were restored after NFκB inhibition and in db(-) /db(-p-50NFκB-/-) and db(-) /db(- P09874 -/-) mice . CONCLUSIONS : The present results indicate that in male type 2 diabetic mice , the augmented NFκB activity also impairs conductance artery function through P09874 and P35354 -dependent mechanisms . Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde-3-phosphate dehydrogenase ( P04406 ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 , CYP2B1/2 , P05181 , CYP3A , catechol-O-methyltransferase ( P21964 ) and microsomal glutathione transferase 1 ( P10620 ) . All but actin and soluble P21964 were positively detected at ≥1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 . The binding correlated well with inhibition of CYP activities , except for P05181 whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 antibodies showed a significant reduction in binding at ≥1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults . TEI-9063 , a stable and highly specific prostacyclin analogue for the prostacyclin receptor in mastocytoma P-815 cells . The prostacyclin ( DB01240 ) analogues , TEI-9063 and its methyl ester , TEI-1324 , have been compared with another stable analogue , iloprost , with respect to binding to the P43119 , stimulation of adenylate cyclase activity and inhibition of thrombin-induced Ca2+ mobilization in mastocytoma P-815 cells . TEI-9063 displaced the [3H]iloprost binding to the membrane fraction , the IC50 value being 3 nM , but showed very low affinity for the PGE receptor . TEI-9063 dose dependently stimulated DB02527 formation in the cells and GTP-dependent adenylate cyclase activity in the membrane fraction , the EC50 value being 50 and 10 nM , respectively . Furthermore , TEI-9063 prevented the thrombin-induced increase in the intracellular Ca2+ concentration , the IC50 value being 50 nM . These IC50 and EC50 values are lower than those obtained for iloprost . On the other hand , those of TEI-1324 were about two-orders higher . Although DB01240 lost its ability to stimulate DB02527 formation by preincubation for 20 min at 37 degrees C , TEI-9063 completely retained its ability after 60-min preincubation . These results demonstrate that TEI-9063 is a stable and stronger agonist for the P43119 than iloprost , and that it prevents thrombin-induced Ca2+ mobilization through stimulation of the adenylate cyclase system in mastocytoma cells . Prostaglandin receptor signaling in disease . Prostanoids , consisting of the prostaglandins ( PGs ) and the thromboxanes ( TXs ) , are a group of lipid mediators formed in response to various stimuli . They include PGD2 , DB00917 , PGF2alpha , DB01240 , and TXA2 . They are released outside of the cells immediately after synthesis , and exert their actions by binding to a G-protein coupled rhodopsin-type receptor on the surface of target cells . There are eight types of the prostanoid receptors conserved in mammals from mouse to human . They are the Q13258 ( DP ) , four subtypes of the PGE receptor ( EP1 , EP2 , EP3 , and EP4 ) , the P43088 ( FP ) , P43119 ( IP ) , and TXA receptor ( TP ) . Recently , mice deficient in each of these prostanoid receptors were generated and subjected to various experimental models of disease . These studies have revealed the roles of PG receptor signaling in various pathological conditions , and suggest that selective manipulation of the prostanoid receptors may be beneficial in treatment of the pathological conditions . Here we review these recent findings of roles of prostanoid receptor signaling and their therapeutic implications . Developmental regulation of prostacyclin synthase and prostacyclin receptors in the ovine uterus and conceptus during the peri-implantation period . This study documents the expression of prostacyclin ( DB01240 ) synthase ( Q16647 ) and DB01240 receptors in the trophoblast and uterus of the ewe at the time of maternal recognition of pregnancy ( i.e. days 7 , 9 , 12 , 14 and 17 ) . The membrane receptor for DB01240 ( P43119 ) and the nuclear receptors , i.e. peroxisome proliferator-activated receptors ( Q07869 ) and their heterodimer partners the retinoid X receptors ( RXR ) , were analysed . In the endometrium , Q16647 transcript and protein were expressed at day 9 of pregnancy and levels declined from days 12 to 17 . Immunohistochemistry and in situ hybridization indicated that Q16647 was mainly located in the luminal epithelium of the endometrium . Endometrial P43119 , Q07869 , P37231 and P48443 expression was regulated during the peri-implantation period whereas Q03181 , P19793 and P28702 were consistently expressed . In the trophoblast , Q16647 transcript levels rose as development progressed and peaked at day 17 . P43119 and Q07869 transcripts peaked before day 12 and then declined and became nearly undetectable by day 17 , whereas Q03181 and P37231 transcript levels rose steadily from days 12 to 17 . Because the PPARs and the RXRs display different expression profiles , we suggest that different heterodimers may form and support distinct functions as development proceeds . Our results also underline the importance of Q16647 and Q03181 in the trophoblast and P43119 in the uterus , suggesting that DB01240 is of both uterine and trophoblastic origin and is involved in a complex signalling pathway at around the time of implantation in the ewe . Beneficial vasoactive endothelial effects of fluvastatin : focus on prostacyclin and nitric oxide . Statins are believed to exert beneficial effects against cardiovascular disease beyond correction of dyslipidemia . There are however still very sparse data on how individual statins interact with the production of vasoactive eicosanoids and nitric oxide ( NO ) in human vascular endothelial cells . Here we have determined how fluvastatin affects the mRNA expression of genes associated with vascular reactivity as well as the formation of two major vasodilators , prostacyclin ( DB01240 ) and NO , in human endothelial cells . Also , the influence of fluvastatin on arterial resistance was assessed in isolated small arteries . We show that the promoter activity of prostacyclin synthase ( Q16647 ) , the mRNA expression of Q16647 and endothelial nitric oxide synthase ( P29474 ) , and the production of DB01240 and NO are significantly induced by fluvastatin . Also , strong rapid dilatation ex vivo was observed , with the equal contribution of DB01240 and NO . Our findings in cell culture experiments and in isolated human arteries indicate that fluvastatin-evoked endothelium-derived vasodilator production may confer protection of the endothelial cells via both acute and long-term effects of fluvastatin treatment . If these effects take place in vivo , we suggest a protective pleiotropic role of fluvastatin on the cardiovascular system , particularly at the level of the vascular endothelium , to ameliorate the process of atherogenesis and in the acute manner to reduce vascular tone . Platelet P43119 affinity is reduced in pre-eclampsia . DB01240 ( DB01240 ) receptors were studied in platelet membrane preparations from women with normal pregnancy , pregnancy-induced hypertension ( PIH ) or pre-eclampsia . Patient groups showed no differences in gestational week at delivery . A markedly lower birth weight , however , was found in pre-eclampsia . No differences between groups could be detected in platelet P43119 number . In contrast , the binding affinity to the DB01240 mimetic iloprost was considerably reduced in pre-eclampsia , whereas receptor affinity between PIH and normal pregnancy did not differ significantly . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Fluorescence imaging reveals the nuclear behavior of peroxisome proliferator-activated receptor/retinoid X receptor heterodimers in the absence and presence of ligand . In a global approach combining fluorescence recovery after photobleaching ( P42345 ) , fluorescence correlation spectroscopy ( FCS ) , and fluorescence resonance energy transfer ( FRET ) , we address the behavior in living cells of the peroxisome proliferator-activated receptors ( PPARs ) , a family of nuclear receptors involved in lipid and glucose metabolism , inflammation control , and wound healing . We first demonstrate that unlike several other nuclear receptors , PPARs do not form speckles upon ligand activation . The subnuclear structures that may be observed under some experimental conditions result from overexpression of the protein and our immunolabeling experiments suggest that these structures are subjected to degradation by the proteasome . Interestingly and in contrast to a general assumption , PPARs readily heterodimerize with retinoid X receptor ( RXR ) in the absence of ligand in living cells . Q07869 diffusion coefficients indicate that all the receptors are engaged in complexes of very high molecular masses and/or interact with relatively immobile nuclear components . PPARs are not immobilized by ligand binding . However , they exhibit a ligand-induced reduction of mobility , probably due to enhanced interactions with cofactors and/or chromatin . Our study draws attention to the limitations and pitfalls of fluorescent chimera imaging and demonstrates the usefulness of the combination of FCS , P42345 , and FRET to assess the behavior of nuclear receptors and their mode of action in living cells . Candidate genetic markers and the risk of restenosis after coronary angioplasty . The aim of the present study was to test for possible associations between candidate gene polymorphisms and the risk of restenosis and recurrent restenosis after percutaneous transluminal coronary angioplasty ( PTCA ) without stenting . We followed up 511 PTCA patients , and restenosis and recurrent restenosis were defined according to angiographical criteria . Genotyping of the beta-fibrinogen -455 G/A , glycoprotein ( GP ) IIIa PlA1/PlA2 , plasminogen activator inhibitor-1 ( P05121 ) 4G/5G , factor V Leiden 1691 G/A , tumour necrosis factor alpha ( TNFalpha ) -238 G/A , TNFalpha -308 G/A , interleukin ( IL ) -1alpha -889 C/T , IL-1beta -511 C/T , methylenetetrahydrofolate reductase ( P42898 ) 677 C/T and endothelial nitric oxide synthase ( P29474 ) 4 b/a gene polymorphisms was performed by PCR and restriction-fragment-length-polymorphism-based techniques . One hundred and sixty patients ( 31.3 % ) developed restenosis and in 130 of these patients , of whom 123 were available for analysis , a second PTCA without stenting was performed . Of these patients , 35 ( 28.5 % ) developed recurrent restenosis . None of the investigated genotypes were associated with the risk of restenosis or recurrent restenosis after PTCA . The degree of stenosis before and immediately after PTCA and the severity of the lesion were independent predictors for restenosis after PTCA . In conclusion , there was no association between the beta-fibrinogen -455 G/A , GP IIIa PlA1/A2 , P05121 4G/5G , factor V Leiden 1691 G/A , TNFalpha -238 G/A , TNFalpha -308 G/A , IL-1alpha -889 C/T , the IL-1beta -511 C/T , P42898 677 C/T and P29474 4 b/a gene polymorphisms and the risk of restenosis after PTCA as well as recurrent restenosis after repeated PTCA . P43119 -induced P40763 phosphorylation in human erythroleukemia cells is mediated via Galpha(s) and Galpha(16) hybrid signaling . Human prostacyclin receptor ( hIP ) stimulates P40763 via pertussis toxin-insensitive G proteins in human erythroleukemia ( HEL ) cells . Since hIP can utilize G(s) and G(q) proteins for signal transduction and that both G proteins can induce P40763 phosphorylation and activation via complex signaling networks , we sought to determine if one of them is predominant in mediating the hIP signal . Stimulation of P40763 DB00135 (705) and DB00133 (727) phosphorylations by the IP-specific agonist , cicaprost , was sensitive to inhibition of protein kinase A , phospholipase Cbeta , protein kinase C , calmodulin-dependent protein kinase II and O60674 /3 . Unlike Galpha(16)-mediated regulation of P40763 in the same cells , cicaprost-induced P40763 DB00135 (705) phosphorylation was resistant to inhibition of Src and MEK while P40763 DB00133 (727) phosphorylation distinctly required phosphatidylinositol-3 kinase . This unique inhibitor-sensitivity pattern of P40763 phosphorylation was reproduced in HEL cells by stimulating the G(16)-coupled C5a receptor in the presence of dibutyryl- DB02527 , suggesting that the change in inhibitor-sensitivity was due to activation of the G(s) pathway . This postulation was confirmed by expressing constitutively active Galpha(16)QL and Galpha(s)QL in human embryonic kidney 293 cells and the inhibitor-sensitivity of Galpha(16)QL-induced P40763 phosphorylations could be converted by the mere presence of Galpha(s)QL to resemble that obtained with cicaprost in HEL cells . In addition , the restoration of the Galpha(16)-mediated inhibitor-sensitivity upon cicaprost induction in Galpha(s)-knocked down HEL cells again verified the pivotal role of G(s) signal . Taken together , our observations illustrate that co-stimulation of G(s) and G(q) can result in the fine-tuning of P40763 activation status , and this may provide the basis for cell type-specific responses following activation of hIP . Impairment of breast cancer cell invasion by P35354 -specific inhibitor NS398 : roles of P61073 and of uPA system . Inhibition of cyclooxygenase-2 ( P35354 ) is known to impair cancer cell metastatic behaviour , but the mechanisms involved largely remain elusive . We aimed to analyse whether the antimetastatic effect of P35354 inhibition in breast cancer cells could be explained by variations in the expression levels of chemokine receptor P61073 , vascular endothelium growth factor ( P15692 ) and Q96NZ9 / Q03405 components of the urokinase plasminogen activator system ( Q03405 ) . Breast cancer cell line MDA-MB-231 was exposed to P35354 -specific inhibitor NS398 . Experimental data were assessed using Matrigel invasion tests , qRT-PCR , ELISA , flow cytometry and MTT test . Exposure to NS398 had no major effect on cell viability , apoptosis or P15692 production . Cell invasion was significantly decreased with reductions ranging from of 3.6 % with 10 μM NS398 to 81.04 % with 100 μM NS398 . P61073 membrane expression was significantly reduced by 18 % ( P < 0.05 ) when cells were treated with 100 μM of NS398 for 72 h . Q96NZ9 mRNA levels were significantly reduced to 78 and 63 % after treatment with 10 μM NS398 for 48 and 72 h , respectively ( P < 0.05 ) . Q03405 mRNA levels also decreased with mild NS398 concentrations , reaching the lowest level of 56 % with 50 μM of NS398 for 48 h ( P < 0.05 ) . With NS398 higher concentrations , Q03405 and Q96NZ9 expression levels increased . According to our results , impairment of expression of P61073 , Q96NZ9 and Q03405 differentially contribute to the antimetastatic effect of P35354 inhibitors depending on drug concentration . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Identification of antithrombin-modulating genes . Role of O95461 , a gene encoding a bifunctional glycosyltransferase , in the secretion of proteins ? The haemostatic relevance of antithrombin together with the low genetic variability of P01008 , and the high heritability of plasma levels encourage the search for modulating genes . We used a hypothesis-free approach to identify these genes , evaluating associations between plasma antithrombin and 307,984 polymorphisms in the GAIT study ( 352 individuals from 21 Spanish families ) . Despite no SNP reaching the genome wide significance threshold , we verified milder positive associations in 307 blood donors from a different cohort . This validation study suggested O95461 , a gene encoding a protein with xylosyltransferase and glucuronyltransferase activities that forms heparin-like linear polysaccharides , as a potential modulator of antithrombin based on the significant association of one SNPs , rs762057 , with anti-FXa activity , particularly after adjustment for age , sex and P01008 rs2227589 genotype , all factors influencing antithrombin levels ( p = 0.02 ) . Additional results sustained this association . O95461 silencing inHepG2 and P29320 -EBNA cells did not affect P01008 mRNA levels but significantly reduced the secretion of antithrombin with moderate intracellular retention . Milder effects were observed on α1-antitrypsin , prothrombin and transferrin . Our study suggests O95461 as the first known modifier of plasma antithrombin , and proposes a new role for O95461 in modulating extracellular secretion of certain glycoproteins . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . Exploration of the antiplatelet activity profile of betulinic acid on human platelets . Betulinic acid , a natural pentacyclic triterpene acid , presents a diverse mode of biological actions including antiretroviral , antibacterial , antimalarial , and anti-inflammatory activities . The potency of betulinic acid as an inhibitor of human platelet activation was evaluated , and its antiplatelet profile against in vitro platelet aggregation , induced by several platelet agonists ( adenosine diphosphate , thrombin receptor activator peptide-14 , and arachidonic acid ) , was explored . Flow cytometric analysis was performed to examine the effect of betulinic acid on P16109 membrane expression and O95456 binding to activated platelets . Betulinic acid potently inhibits platelet aggregation and also reduced O95456 binding and the membrane expression of P16109 . Principal component analysis was used to screen , on the chemical property space , for potential common pharmacophores of betulinic acid with approved antithrombotic drugs . A common pharmacophore was defined between the NMR-derived structure of betulinic acid and prostacyclin agonists ( DB01240 ) , and the importance of its carboxylate group in its antiplatelet activity was determined . The present results indicate that betulinic acid has potential use as an antithrombotic compound and suggest that the mechanism underlying the antiplatelet effects of betulinic acid is similar to that of the P43119 agonists , a hypothesis that deserves further investigation . P35354 -derived prostacyclin confers atheroprotection on female mice . Female gender affords relative protection from cardiovascular disease until the menopause . We report that estrogen acts on estrogen receptor subtype alpha to up-regulate the production of atheroprotective prostacyclin , DB01240 , by activation of cyclooxygenase 2 ( P35354 ) . This mechanism restrained both oxidant stress and platelet activation that contribute to atherogenesis in female mice . Deletion of the P43119 removed the atheroprotective effect of estrogen in ovariectomized female mice . This suggests that chronic treatment of patients with selective inhibitors of P35354 could undermine protection from cardiovascular disease in premenopausal females . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . On the relevance of glycolysis process on brain gliomas . The proposed analysis considers aspects of both statistical and biological validation of the glycolysis effect on brain gliomas , at both genomic and metabolic level . In particular , two independent datasets are analyzed in parallel , one engaging genomic ( Microarray Expression ) data and the other metabolomic ( Magnetic Resonance Spectroscopy Imaging ) data . The aim of this study is twofold . First to show that , apart from the already studied genes ( markers ) , other genes such as those involved in the human cell glycolysis significantly contribute in gliomas discrimination . Second , to demonstrate how the glycolysis process can open new ways towards the design of patient-specific therapeutic protocols . The results of our analysis demonstrate that the combination of genes participating in the glycolytic process ( P04075 , P09972 , P09104 , P04406 , P52789 , P00338 , P07195 , P40925 , P11177 , P08237 , P06744 , P00558 , P36871 and P30613 ) with the already known tumor suppressors ( P60484 , Rb , P04637 ) , oncogenes ( P11802 , P00533 , PDGF ) and Q9BYW2 , enhance the discrimination of low versus high-grade gliomas providing high prediction ability in a cross-validated framework . Following these results and supported by the biological effect of glycolytic genes on cancer cells , we address the study of glycolysis for the development of new treatment protocols . Carbacyclin induces carnitine palmitoyltransferase-1 in cardiomyocytes via peroxisome proliferator-activated receptor ( Q07869 ) delta independent of the IP receptor signaling pathway . DB01240 ( DB01240 ) and its analogues exert cardioprotective effects via the rhodopsin type membrane P43119 , IP . Peroxisome proliferator-activated receptor ( Q07869 ) delta is a nuclear receptor abundantly expressed in cardiomyocytes and plays a pivotal role in maintaining constitutive mitochondrial fatty acid beta-oxidation ( FAO ) . Recently , a novel signaling pathway of DB01240 via PPARdelta has been demonstrated in non-cardiac tissues . We therefore examined whether carbacyclin ( cPGI2 ) , a DB01240 analogue , up-regulates transcriptional expression of carnitine palmitoyltransferase-1 ( CPT-1 ) , the rate-limiting enzyme in mitochondrial FAO , via PPARdelta in cardiomyocytes . Intraperitoneal injection of cPGI2 increased CPT-1 mRNA expression in murine hearts . Transcriptional activity was evaluated by Q07869 responsive element ( PPRE ) -luciferase reporter gene assay in cultured neonatal rat cardiomyocytes . CPT-1 mRNA expression and PPRE promoter activity were significantly increased by cPGI2 in a concentration-dependent manner , where PPRE has been mapped to the promoter region of the CPT-1 gene . Moreover , the elevation of CPT-1 mRNA expression and PPRE promoter activity by cPGI2 was not abolished by H-89 , a potent protein kinase A inhibitor , but was significantly inhibited in cardiomyocytes over-expressing a dominant-negative type of PPARdelta . Furthermore , electrophoretic mobility shift assays demonstrated that binding of PPARdelta to PPRE in the CPT-1 gene promoter is enhanced in response to cPGI2 stimulation . In addition , down-regulation of CPT-1 mRNA expression in cardiomyocytes subjected to hypoxia was attenuated by cPGI2 . These results indicate that cPGI2 induces CPT-1 mRNA expression through PPARdelta , independent of the IP receptor signaling pathway , suggesting a possibility that cPGI2 modulates cardiac energy metabolism by activating FAO via PPARdelta . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Inhibition of myoblast migration by prostacyclin is associated with enhanced cell fusion . Satellite cells are stem cells that are critical for the formation and growth of skeletal muscle during myogenesis . To differentiate and fuse , proliferating satellite cells or myoblasts must migrate and establish stable cell-cell contacts . However , the factors that regulate myoblast migration and fusion are not understood completely . We have identified DB01240 as a novel regulator of myogenesis in vitro . DB01240 is a member of the family of prostaglandins ( PG ) , autocrine/paracrine signaling molecules synthesized via the cyclooxygenase-1 and -2 pathways . Primary mouse muscle cells both secrete DB01240 and express the P43119 , IP , at various stages of myogenesis . Using genetic and pharmacological approaches , we show that DB01240 is a negative regulator of myoblast migration that also enhances cell fusion . Thus , DB01240 may act as a " brake " on migrating cells to facilitate cell-cell contact and fusion . Together , our results highlight the importance of the balance between positive and negative regulators in cell migration and myogenesis . This work may have implications for migration of other populations of adult stem cells and/or cells that undergo fusion . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . DB01240 -IP signaling and prostaglandin E2-EP2/EP4 signaling both mediate joint inflammation in mouse collagen-induced arthritis . Prostaglandin (PG)I2 ( prostacyclin [ P06744 ] ) and DB00917 are abundantly present in the synovial fluid of rheumatoid arthritis ( RA ) patients . Although the role of DB00917 in RA has been well studied , how much DB01240 contributes to RA is little known . To examine this issue , we backcrossed mice lacking the P43119 ( IP ) to the DBA/1J strain and subjected them to collagen-induced arthritis ( CIA ) . IP-deficient ( IP-/- ) mice exhibited significant reduction in arthritic scores compared with wild-type ( WT ) mice , despite anti-collagen antibody production and complement activation similar to WT mice . IP-/- mice also showed significant reduction in contents of proinflammatory cytokines , such as interleukin ( IL ) -6 in arthritic paws . Consistently , the addition of an IP agonist to cultured synovial fibroblasts significantly enhanced P05231 production and induced expression of other arthritis-related genes . On the other hand , loss or inhibition of each PGE receptor subtype alone did not affect elicitation of inflammation in CIA . However , a partial but significant suppression of CIA was achieved by the combined inhibition of EP2 and EP4 . Our results show significant roles of both DB01240 -IP and DB00917 -EP2/EP4 signaling in the development of CIA , and suggest that inhibition of DB00917 synthesis alone may not be sufficient for suppression of RA symptoms . Enhanced NF-κB activity impairs vascular function through P09874 - , SP-1- , and P35354 -dependent mechanisms in type 2 diabetes . Type 2 diabetes ( T2D ) is associated with vascular dysfunction . We hypothesized that increased nuclear factor-κB ( NF-κB ) signaling contributes to vascular dysfunction in T2D . We treated type 2 diabetic ( db(-)/db(-) ) and control ( db(-)/db(+) ) mice with two NF-κB inhibitors ( 6 mg/kg dehydroxymethylepoxyquinomicin twice a week and 500 μg/kg/day IKK-NBD peptide ) for 4 weeks . Pressure-induced myogenic tone was significantly potentiated , while endothelium-dependent relaxation ( EDR ) was impaired in small coronary arterioles and mesenteric resistance artery from diabetic mice compared with controls . Interestingly , diabetic mice treated with NF-κB inhibitors had significantly reduced myogenic tone potentiation and improved EDR . Importantly , vascular function was also rescued in db(-)/db(-p50NF-κB-/-) and db(-)/db(- P09874 -/-) double knockout mice compared with db(-)/db(-) mice . Additionally , the acute in vitro downregulation of NF-κB-p65 using p65NF-κB short hairpin RNA lentivirus in arteries from db(-)/db(-) mice also improved vascular function . The NF-κB inhibition did not affect blood glucose level or body weight . The RNA levels for Sp-1 and P29474 phosphorylation were decreased , while p65NF-κB phosphorylation , cleaved poly(ADP-ribose) polymerase ( PARP ) -1 , and cyclooxygenase ( P36551 ) -2 expression were increased in arteries from diabetic mice , which were restored after NF-κB inhibition and in db(-)/db(-p50NF-κB-/-) and db(-)/db(- P09874 -/-) mice . In the current study , we provided evidence that enhanced NF-κB activity impairs vascular function by P09874 - , Sp-1- , and P35354 -dependent mechanisms in male type 2 diabetic mice . Therefore , NF-κB could be a potential target to overcome diabetes-induced vascular dysfunction . Amelioration of nephropathy with apoA-1 mimetic peptide in apoE-deficient mice . BACKGROUND : There is mounting evidence that dyslipidaemia may contribute to development and progression of renal disease . For instance , hyperlipidaemia in apolipoprotein E-deficient ( apoE(-/-) ) mice is associated with glomerular inflammation , mesangial expansion and foam cell formation . ApoA-1 mimetic peptides are potent antioxidant and anti-inflammatory compounds which are highly effective in ameliorating atherosclerosis and inflammation in experimental animals . Given the central role of oxidative stress and inflammation in progression of renal disease , we hypothesized that apoA-1 mimetic peptide , D-4F , may attenuate renal lesions in apoE(-/-) mice . METHODS : Twenty-five-month-old female apoE(-/-) mice were treated with D-4F ( 300 µg/mL in drinking water ) or placebo for 6 weeks . Kidneys were harvested and examined for histological and biochemical characteristics . RESULTS : Compared with the control mice , apoE(-/-) mice showed significant proteinuria , tubulo-interstitial inflammation , mesangial expansion , foam cell formation and up-regulation of oxidative [ NAD(P)H oxidase subunits ] and inflammatory [ NF-κB , P13500 , P05121 and P35354 ] pathways . D-4F administration lowered proteinuria , improved renal histology and reversed up-regulation of inflammatory and oxidative pathways with only minimal changes in plasma lipid levels . CONCLUSIONS : The apoE(-/-) mice develop proteinuria and glomerular and tubulo-interstitial injury which are associated with up-regulation of oxidative and inflammatory mediators in the kidney and are ameliorated by the administration of apoA-1 mimetic peptide . These observations point to the role of oxidative stress and inflammation in the pathogenesis of renal disease in hyperlipidaemic animals and perhaps humans . Expression of prostacyclin receptors in luteinizing hormone-releasing hormone immortalized neurons : role in the control of hormone secretion . PGs of the E series are involved in the control of P01148 secretion . The present experiments were conducted to clarify whether DB01240 ( prostacyclin ) might be also involved in such a control , using multiple methodological approaches on immortalized P01148 -secreting neurons . A RT-PCR procedure to detect mouse P43119 ( IP ) messenger RNA was first applied , and the results obtained showed the presence of a specific transcript in two cell lines of immortalized P01148 neurons ( GT1-1 and GN11 cell lines ) . Receptor binding assays on membrane preparations from GT1-1 cells showed the presence of a single specific and saturable class of binding sites ( Kd = 4.6 nM ; 10,000 sites/cell ) for [3H]iloprost , a stable analog of DB01240 . Competition experiments showed that the binding sites labeled by [3H]iloprost possess the pharmacological characteristics of IP receptors . In functional studies , DB01240 and its analogs , iloprost and cicaprost , were able to stimulate P01148 release from the GT1-1 cells with elevated potencies ( EC50 = 0.6-4.3 nM ) ; PGE1 was only slightly less active ( EC50 = 28.5 nM ) , whereas DB00917 , considered the major PG involved in P01148 secretion , was poorly effective ( EC50 = 921 nM ) . The relative potencies ( EC50 ) of these compounds in stimulating the intracellular accumulation of DB02527 were in line with their P01148 -releasing activities . In conclusion , these results indicate that immortalized P01148 -secreting neurons express IP receptors through which DB01240 may exert relevant effects on P01148 release . Cellular and molecular biology of prostacyclin synthase . Q16647 ( PGIS ) cDNA comprises 1500 nucleotides coding for a 500 amino acid protein . It is a heme protein with spectral characteristics of cytochrome p450 ( CYP ) . It does not possess the typical CYP mono-oxygenase activity but catalyzes the rearrangement of prostaglandin H2 to form DB01240 . Analysis of its structure-function by molecular modeling and site-directed mutagenesis reveals a long substrate channel lined by hydrophobic residues . DB00151 -441 has been identified as the proximal axial ligand of heme . DB00135 -430 is nitrated by peroxynitrite which results in reduced PGIS catalytic activity , suggesting that DB00135 -430 is located close to the heme pocket . PGIS is constitutively expressed and may be upregulated by cytokines , reproductive hormones , and growth factors . It is physically colocalized with cyclooxygenases and phospholipases , and functionally coupled with these enzymes . PGIS coupling with P35354 has been shown to play an important role in vascular protection , embryo development and implantation , and cancer growth . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Morphologic changes in explanted lungs after prostacyclin therapy for pulmonary hypertension . DB01240 ( DB01240 ) causes vasodilation and inhibition of platelet aggregation in vivo . DB01240 is also postulated to affect pulmonary vascular remodeling , at least partly through anti-proliferative effect via P43119 ( PGIR ) . However , the mechanism(s) of action by which ( DB01240 ) exerts its therapeutic effect is still not clear despite clear clinical benefit seen in severe pulmonary hypertension ( PH ) patients . We performed a histopathologic and morphometric study on the explanted lung tissues from DB01240 -treated patients prior to lung transplantation ( n = 9 ) , in an attempt to elucidate morphologic changes associated with DB01240 treatment . Explanted lungs from PH patients without DB01240 treatment were examined as the control ( n = 11 ) . We also studied the possible differences in PGIR expression between the treated and untreated groups by immunohistochemical method . Seven out of 9 treated patients showed moderate to severe bronchial and perivascular inflammation , as opposed to only 1 such case in the control group . Five out of 9 treated cases showed moderate to severe alveolar edema with or without evidence of old hemorrhage , in contrast to only 1 case showing moderate alveolar edema in control patients . Morphometry did not reveal any significant difference between the two groups either in the % thickness of intima , media , or adventitia or in the density of plexiform lesions . Immunostain also failed to demonstrate any notable difference in PGIR expression . In conclusion , DB01240 -treated cases revealed more pronounced pulmonary alveolar edema and inflammation , but no morphological evidence of altered vascular remodeling or PGIR expression after DB01240 therapy . Dual P00533 and P42345 targeting in squamous cell carcinoma models , and development of early markers of efficacy . The epidermal growth factor receptor ( P00533 ) is a validated target in squamous cell carcinoma ( SCC ) of the head and neck . Most patients , however , do not respond or develop resistance to this agent . P42345 ( P42345 ) is involved in the pathogenesis of SCC of the head and neck ( SCCHN ) . This study aimed to determine if targeting P42345 in combination with P00533 is effective in SCC , and to develop early pharmacodynamic markers of efficacy . Two SCC cell lines , one resistant ( HEP2 ) and one of intermediate susceptibility ( Detroit 562 ) to P00533 inhibitors , were xenografted in vivo and treated with an P42345 inhibitor ( temsirolimus ) , an P00533 inhibitor ( erlotinib ) or a combination of both . DB06287 exerted superior growth arrest in both cell lines than erlotinib . The combined treatment resulted in synergistic antitumor effects in the Detroit 562 cell line . Immunohistochemical assessment of pharmacodynamic effects in fine-needle aspiration ( FNA ) biopsies early after treatment using phospho MAPK , Phospho-P70 and Ki67 as end points demonstrated pathway abrogation in the Detroit 562 tumours treated with the combination , the only group where regressions were seen . In conclusion , an P42345 inhibitor showed antitumor activity in P00533 -resistant SCC cell lines . Marked antitumor effects were associated with dual pathway inhibition , which were detected by early FNA biopsies . Gene expression studies provide clues to the pathogenesis of uterine leiomyoma : new evidence and a systematic review . BACKGROUND : Uterine leiomyomas are extremely common and a major cause of pelvic pain , bleeding , infertility , and the leading indication for hysterectomy . Familial and epidemiological studies provide compelling evidence that genetic alterations play an important role in leiomyoma development . METHODS : Using Affymetrix U133A GeneChip we analysed expression profiles of 22,283 genes in paired samples of leiomyoma and adjacent normal myometrium . We compared our results with previously published data on gene expression in uterine leiomyoma and identified the overlapping gene alterations . RESULTS : We detected 80 genes with average differences of > or = 2-fold and false discovery rates of < 5 % ( 14 overexpressed and 66 underexpressed ) . A comparative analysis including eight previous gene expression studies revealed eight prominent genes ( P07327 , P18847 , P29373 , O00622 , Q07507 , P42262 , P01344 , Q5EB52 ) identified by at least five different studies , eleven genes ( P00352 , P25063 , P29279 , O43602 , P28562 , P01100 , O60829 , P24592 , P41222 , P43115 , P04818 ) reported by four studies , twelve genes ( ABCA , P04083 , Q15847 , O00585 , P38936 , Q14194 , P54849 , P03372 , FY , Q99683 , P37173 , P35625 ) identified by three studies , and 40 genes reported by two different studies . CONCLUSIONS : Review of gene expression data revealed concordant changes in genes regulating retinoid synthesis , IGF metabolism , TGF-beta signaling and extracellular matrix formation . Gene expression studies provide clues to the relevant pathways of leiomyoma development . The anticoagulant effect of PGI2S and tPA in transgenic umbilical vein endothelial cells is linked to up-regulation of PKA and PKC . The selection of vascular grafts for coronary artery bypass surgery is crucial for a positive outcome . This study aimed to establish a novel line of vascular endothelial cells with a potent anticoagulant effect . A lentiviral vector was used to stably transfect human umbilical vein endothelial cells ( HUVECs ) with PGI2S alone ( HUVEC-PGI2S ) or both PGI2S and tPA ( HUVEC-PGI2S-tPA ) . Both HUVEC-PGI2S and HUVEC-PGI2S-tPA cells over-expressing PGI2S and tPA were compared to mock-transfected cells . The enzyme-linked immuno sorbent assay ( ELISAs ) demonstrated that the anticoagulation components , P01008 and P00747 , were up-regulated and coagulation factor FVIII was down-regulated in both cell lines . QRT-PCR and western blotting demonstrated the vasodilation and platelet disaggregation proteins PKA , PKC , and P43119 were up-regulated in both cell lines , but MAPK expression was not altered in either cell line . However , cell viability and colony formation assays and cell cycle analysis demonstrated that both cell lines had a lower rate of cell growth and induced P55008 phase arrest . HUVEC-PGI2S and HUVEC-PGI2S-tPA cells have a potent anticoagulant effect and their use in vascular heterografts may decrease the risk of thrombosis . Q16647 gene transfer inhibits neointimal formation by suppressing Q07869 delta expression . OBJECTIVE : DB01240 ( P06744 (2) ) is a potent ligand of peroxisome proliferator-activated receptor delta ( Q07869 delta ) that regulates cell growth and differentiation . The aim of this study was to elucidate how endogenous P06744 (2) overexpression affects the expressions of Q07869 delta and mitogen-activated protein kinases ( MAPKs ) in the development of neointimal formation in experimental angioplasty with adenovirus-mediated P06744 (2) synthase ( Ad-PGIS ) gene transfer . METHODS AND RESULTS : In human aortic smooth muscle cells , protein blotting analysis showed that P06744 (2) overproduction decreased the levels of phosphorylated p38 MAPK ( P-p38 MAPK ) ( 2.0-fold versus 0.83-fold relative to control ) . Immunohistochemical analysis in balloon-injured arteries revealed diffuse expression of Q07869 delta in the neointima of control vessels , with no expression in uninjured vessels . The level of Q07869 delta expression was lower in Ad-PGIS-treated arteries than in control vessels , with the Q07869 delta localized in the neointima adjacent to endothelium . Staining of P-p38 MAPK showed a similar pattern to Q07869 delta among the three groups . Morphometric analysis at day 14 revealed that Ad-PGIS reduced the intima-to-media ratio by up to 59 % . CONCLUSIONS : Ad-PGIS gene transfer reduced Q07869 delta expression and inhibited neointimal formation after balloon injury in accordance with the reduction in the phosphorylation of p38 MAPK . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Diverse prostaglandin receptors activate distinct signal transduction pathways in rat myometrium . Attempts were made to identify prostaglandin ( PG ) receptors in rat myometrium , according to the differential rank order of potencies displayed by the natural PGs and their analogues , both at the level of second messenger generation and contraction . In estrogen-treated rat myometrium , PGs [ iloprost = DB01240 greater than DB00917 much greater than 16,16-dimethyl ( DM ) - DB00917 ; sulprostone = misoprostol = 0 ] induced adenosine 3',5'-cyclic monophosphate generation , indicating the contribution of a P43119 . The generation of inositol phosphates was stimulated by PGs ( PGF2 alpha greater than PGD2 much greater than DB00917 = DM- DB00917 much greater than iloprost greater than sulprostone = misoprostol = 0 ) , reflecting a PGF2 alpha-receptor-mediated process , which was insensitive to pertussis toxin ( PTX ) . Contractions caused by PGF2 alpha were closely correlated to PGF2 alpha-receptor activation associated with the phospholipase C pathway . By contrast , contractions evoked by DB00917 , equally mimicked by sulprostone and misoprostol , were abolished by PTX and were independent of phospholipase C activation . In the pregnant myometrium ( day 21 ) , the latter PGE-receptor-mediated mechanism also contributed to contractions caused by DB00917 ( less than microM concn ) . Phospholipase C activation was coupled not only to PGF2 alpha but also to PGE receptors and could be correlated with contractions induced by PGF2 alpha and DB00917 greater than microM concn ) . All PGs tested were coupled to inhibitory G protein-mediated adenylate cyclase inhibition , displaying an equipotency that did not allow characterization of the inhibitory PG receptors . Polyamines and membrane transporters . In recent years , our understanding of the importance of membrane transporters ( MTs ) in the disposition of and response to drugs has increased significantly . MTs are proteins that regulate the transport of endogenous molecules and xenobiotics across the cell membrane . In mammals , two super-families have been identified : DB00171 -binding cassette ( DB01048 ) and solute carrier ( O00585 ) transporters . There is evidence that MTs might mediate polyamines ( PA ) transport . PA are ubiquitous polycations which are found in all living cells . In mammalian cells , three major PA are synthesised : putrescine , spermidine and spermine ; whilst the decarboxylated arginine ( agmatine ) is not produced by mammals but is synthesised by plants and bacteria . In addition , research in the PA field suggests that PA are transported into cells via a specific transporter , the polyamine transport system(s) ( Q03393 ) . Although the Q03393 has not been fully defined , there is evidence that some of the known MTs might be involved in PA transport . In this mini review , eight O00585 transporters will be reviewed and their potential to mediate PA transport in human cells discussed . These transporters are O15245 , O15244 , O75751 , Q96FL8 , P30825 , P08195 , SLC12A8A , and Q86VW1 . Preliminary data from our laboratory have revealed that O15245 might be involved in the PA uptake ; in addition to one member of ABC superfamily ( P08183 protein ) might also mediate the efflux of polyamine like molecules .

MH_train_44

MH_train_44

MH_train_44

interacts_with DB08870?

multiple_choice

A comparative assessment of fracture resistance of endodontically treated and re-treated teeth : An in vitro study . AIM : To compare and assess the fracture resistance of endodontically treated teeth with those that have been subjected to endodontic retreatment . MATERIALS AND METHODS : 30 extracted mandibular premolars were decoronated at cementoenamel junction and randomly divided into 2 groups . In Group I endodontic treatment was performed with ProTaper rotary system till size F2 and obturated . In Group II , cleaning and shaping was done and teeth were subjected to Spiral CT to assess the remaining dentin thickness and obturated . Later retreatment was done using Protaper Universal Retreatment system and final shaping was performed till size P13726 . Remaining dentin thickness was again assessed using Spiral CT and then obturated . All the specimens were subjected to fracture resistance using universal testing machine . The results were statistically analyzed using Independent Samples t-test for analysis of remaining dentin thickness using P09683 within Group II and Paired Samples t-test was used for assessment of fracture resistance between Group I and II ( P < 0.05 ) . RESULTS : In Group II , the intra group comparison of the remaining dentin thickness in the coronal third reveals statistical significance , with a significant difference noted in the apical third . Results of the fracture resistance reveal a statistically significant difference ( P < 0.05 ) between Groups I and II with the mean fracture resistance of Group I being higher than Group II . CONCLUSION : Endodontically retreated teeth have shown significantly decreased resistance to fracture and this has a positive correlation to the increased loss of root dentin during the retreatment procedures . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . P00533 variant III mutations in lung tumorigenesis and sensitivity to tyrosine kinase inhibitors . The tyrosine kinase inhibitors gefitinib ( DB00317 ) and erlotinib ( Tarceva ) have shown anti-tumor activity in the treatment of non-small cell lung cancer ( NSCLC ) . Dramatic and durable responses have occurred in NSCLC tumors with mutations in the tyrosine kinase domain of the epidermal growth factor receptor ( P00533 ) . In contrast , these inhibitors have shown limited efficacy in glioblastoma , where a distinct P00533 mutation , the variant III ( vIII ) in-frame deletion of exons 2-7 , is commonly found . In this study , we determined that EGFRvIII mutation was present in 5 % ( 3/56 ) of analyzed human lung squamous cell carcinoma ( SCC ) but was not present in human lung adenocarcinoma ( 0/123 ) . We analyzed the role of the EGFRvIII mutation in lung tumorigenesis and its response to tyrosine kinase inhibition . Tissue-specific expression of EGFRvIII in the murine lung led to the development of NSCLC . Most importantly , these lung tumors depend on EGFRvIII expression for maintenance . Treatment with an irreversible P00533 inhibitor , HKI-272 , dramatically reduced the size of these EGFRvIII-driven murine tumors in 1 week . Similarly , Ba/ P13726 cells transformed with the EGFRvIII mutant were relatively resistant to gefitinib and erlotinib in vitro but proved sensitive to HKI-272 . These findings suggest a therapeutic strategy for cancers harboring the EGFRvIII mutation . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . Rescue from failed growth factor and/or chemotherapy P19526 mobilization with G- P04141 and plerixafor ( DB06809 ) : an institutional experience . Auto- P09683 has been shown to be a potentially curative treatment for a variety of hematological malignancies . Auto- P09683 is dependent on the successful mobilization and collection of hematopoietic stem cells to ensure engraftment . The inability to mobilize sufficient number of hematopoietic stem cells using standard cytokine-assisted mobilization strategies excludes eligible patients from potentially curative auto- P09683 . DB06809 ( DB06809 ; DB06809 ) , a novel bicyclam antagonist of the SDF-1alpha/ P61073 complex , has been reported previously to augment PBSC mobilization in patients undergoing their first planned stem cell mobilization and collection attempt . In our experience , 17 of 20 patients otherwise eligible for auto- P09683 who failed previous mobilization attempts had successful mobilization of P28906 (+) hematopoietic stem cells with one apheresis procedure , and an additional patient required two aphereses procedures , when treated with the combination of plerixafor and G- P04141 on a compassionate use protocol available at our institution . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment . Scutellarein Reduces Inflammatory Responses by Inhibiting Src Kinase Activity . Flavonoids are plant pigments that have been demonstrated to exert various pharmacological effects including anti-cancer , anti-diabetic , anti-atherosclerotic , anti-bacterial , and anti-inflammatory activities . However , the molecular mechanisms in terms of exact target proteins of flavonoids are not fully elucidated yet . In this study , we aimed to evaluate the anti-inflammatory mechanism of scutellarein ( P09683 ) , a flavonoid isolated from Erigeron breviscapus , Clerodendrum phlomidis and Oroxylum indicum Vent that have been traditionally used to treat various inflammatory diseases in China and Brazil . For this purpose , a nitric oxide ( NO ) assay , polymerase chain reaction ( PCR ) , nuclear fractionation , immunoblot analysis , a kinase assay , and an overexpression strategy were employed . Scutellarein significantly inhibited NO production in a dose-dependent manner and reduced the mRNA expression levels of inducible NO synthase ( P35228 ) and tumor necrosis factor ( P01375 ) -α in lipopolysaccharide ( LPS ) -activated RAW264.7 cells . In addition , P09683 also dampened nuclear factor ( NF ) -κB-driven expression of a luciferase reporter gene upon transfection of a TIR-domain-containing adapter-inducing interferon-β ( Q8IUC6 ) construct into Human embryonic kidney 293 ( P29320 293 ) cells ; similarly , NF-κ B nuclear translocation was inhibited by P09683 . Moreover , the phosphorylation levels of various upstream signaling enzymes involved in NF-κB activation were decreased by P09683 treatment in LPS-treated RAW264.7 cells . Finally , P09683 strongly inhibited Src kinase activity and also inhibited the autophosphorylation of overexpressed Src . Therefore , our data suggest that P09683 can block the inflammatory response by directly inhibiting Src kinase activity linked to NF-κB activation . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . DB08870 followed by allogeneic transplantation as salvage regimen in patients with relapsed and/or refractory Hodgkin 's lymphoma . Patients with relapsed or refractory Hodgkin lymphoma ( RR-HL ) have poor outcomes . DB08870 ( BV ) , an antibody-drug conjugate comprising an anti- P28908 antibody conjugated to the potent anti-microtubule agent , monomethyl auristatin E , induces high tumour responses with moderate adverse effects . In a retrospective study , we describe objective response rates and subsequent allogeneic stem cell transplantation ( allo- P09683 ) in patients with RR-HL treated by BV in a named patient program in two French institutions . Twenty-four adult patients with histologically proven P28908 (+) RR-HL treated with BV were included from July 2009 to November 2012 . Response to BV treatment was evaluated after four cycles . Eleven patients were in complete response ( 45.8 % ) , while five patients were in partial response ( 20.8 % ) , with an overall response rate of 66.6 % . Eight patients failed to respond to BV ( 33.3 % ) . All of the responding patients could receive consolidation treatment after BV : three patients underwent autologous stem cell transplantation ( auto- P09683 ) , three patients received a tandem auto- P09683 /allo- P09683 , nine patients received allo- P09683 and one patient was treated with donor lymphocyte infusion . We found no treatment-related mortality at day 100 among the 12 patients who underwent BV following by allogeneic transplantation . With a median follow-up of 20 months ( range 10.5-43.2 ) , none of them relapsed or died . BV followed by allo- P09683 represents an effective salvage regimen in patients with RR-HL . Mutant enrichment with 3'-modified oligonucleotides a practical PCR method for detecting trace mutant DNAs . Many clinical situations necessitate highly sensitive and reliable molecular assays ; however , the achievement of such assays remains a challenge due to the inherent limitations of molecular testing methods . Here , we describe a simple and inexpensive enrichment technique that we call mutant enrichment with 3'-modified oligonucleotides ( Q9Y316 ) . The method is based on the use of a 3'-modified oligonucleotide primer that blocks extension of the normal allele but enables extension of the mutated allele . The performance of the technique was evaluated with respect to its ability to detect common cancer mutations in the P00533 , P01116 , P15056 , P04637 , O60674 , and P06748 genes . We achieved sensitivities of 10(-2) to 10(-6) using downstream Sanger sequencing , depending on the concentrations and thermodynamics of the primers . Q9Y316 may be applicable to the quantitative real-time PCR platform and other downstream assays . This technique may be practically applicable to various medical situations . Fanconi anemia : current management . Fanconi anemia ( FA ) is an autosomal recessive chromosomal instability disorder , characterized by congenital anomalies , defective hematopoiesis and a high risk of developing acute myeloid leukemia and certain solid tumors . All racial and ethnic groups are at risk , and at least 11 complementation groups have been identified and the genes defective in eight of these have been identified ( O15360 , C , D2 , E , F , G , L and P51587 ) . FA-A is the most common complementation group , accounting for approximately 65 % of all affected individuals . The gold-standard screening test for FA is based on the characteristic hypersensitivity of FA cells to the crosslinking agents , such as mitomicin C or diepoxybutane . Recent progress has been made in identifying the genes bearing pathogenetically relevant mutations , but slower progress has been made in defining the precise functions of the proteins in normal cells , in part because that the proteins are multifunctional . Molecular studies have established that a common pathway exist , both between the FA proteins and other proteins involved in DNA repair such as NBS1 , Q13315 , P38398 and P51587 . Stem cell transplantation ( P09683 ) is the only option for establishing normal hematopoiesis . To reduce undue toxicities due to inherent hypersensitivity , nonmyeloablative conditioning for transplants has been advocated . This review summarizes the general clinical and hematologic features and the current management of FA . Fanconi anemia ( FA ) is the commonest type of inherited bone marrow failure syndrome with the birth incidence of around three per million . The inheritance pattern is autosomal recessive with the estimated heterozygote frequency being one in 300 in Europe and the US . P00797 angiotensin system modulates P42345 pathway through AT2R in HIVAN . P42345 ( P42345 ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 and p70S6K . DB09026 , a renin inhibitor attenuated phosphorylation of both P42345 and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 ) phosphorylation of p70S6K in a dose dependent manner . HIV/ P04629 also displayed enhanced phosphorylation of both P42345 and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 and p70S6K . These findings indicate that HIV stimulates P42345 pathway in HIVAN through the activation of renin angiotensin system via AT2R . Intracellular capture of P33681 in antigen-presenting cells reduces costimulatory activity . P16410 gene constructs were designed to express P16410 exclusively in the endoplasmic reticulum ( ER ) . Four different P16410 gene constructs were transfected into P29320 293 ( human embryonic kidney ) and A20 ( Balb/c mouse B lymphoma ) cells . All constructs contained an ER retention signal and coded for P16410 expression in the ER . One of the constructs , which contained the membrane part of P16410 , coded for an expression both on the cell surface and in the ER . Three of the expressed P16410 types ( including the ER-membrane-expressed form ) caused a reduced surface expression of P33681 in the A20 cells . Only constructs which allow dimerization of P16410 showed this effect . It is assumed that intracellular P16410 bound P33681 and inhibited therefore the transport of P33681 to the surface . The binding obviously caused also an enhanced degradation of the complexes because both proteins showed a low concentration in the transfected cell lines . P16410 -transfected and P33681 -reduced A20 cells showed a diminished costimulating activity upon T cells . This was demonstrated by a reduced proliferation of T cells from ovalbumin-immunized Balb/c mice , incubated with ovalbumin peptide-primed P16410 -transfected A20 cells . Effect of acetazolamide on aquaporin-1 and fluid flow in cultured choroid plexus . DB00819 ( AZA ) , used in treatment of early or infantile hydrocephalus , is effective in some cases , while its effect on the choroid plexus ( CP ) remains ill-defined . The drug reversibly inhibits aquaporin-4 ( P55087 ) , the most ubiquitous " water pore " in the brain , and perhaps modulation of P29972 ( located apically on CP cells ) by AZA may reduce cerebrospinal fluid ( P04141 ) production . We sought to elucidate the effect of AZA on P29972 and fluid flow in CP cell cultures.CP tissue culture from 10-day Sprague-Dawley rats and a TRCSF-B cell line were grown on Transwell permeable supports and treated with 100 μM AZA . Fluid assays to assess direction and extent of fluid flow , and P29972 expression patterns by immunoblot , Immuncytochemistry ( ICC ) , and quantitative reverse transcriptase polymerase chain reaction ( qRT-PCR ) were performed.Immunoblots and ICC analyses showed a decrease in P29972 protein shortly after AZA treatment ( lowest at 12 h ) , with transient P29972 reduction mediated by mRNA expression ( lowest at 6 h ) . Transwell fluid assays indicated a fluid shift at 2 h , before significant changes in P29972 mRNA or protein levels.Timing of AZA effect on P29972 suggests the drug alters protein transcription , while affecting fluid flow by a concomitant method . It is plausible that other mechanisms account for these phenomena , as the processes may occur independently . Molecular characterization of de novo Philadelphia chromosome-positive acute myeloid leukemia . Philadelphia chromosome-positive ( Ph+ ) acute myeloid leukemia ( AML ) is a controversial diagnosis , as others propose that it represents chronic myelogenous leukemia in blast phase ( CML-BP ) . P06748 mutations occur in 25-35 % of patients with AML but are absent in patients with CML . Conversely , P00519 mutations occur in 25 % of imatinib-naive patients with CML-BP but are not described in patients with AML . We analyzed for P06748 and P00519 mutations in nine Ph+ patients with AML and five patients with CML-BP initially presenting in BP . In six cases of Ph+ AML , we screened for a panel of gene mutations using Sequenome(®)-based methods including P31749 , P31751 , Q9Y243 , P15056 , P00533 , P50148 , GNAS , O75874 , P48735 , P01116 , MET , P01111 , P42336 and P07949 . Two of nine ( 22 % ) patients with Ph+ AML had P06748 mutations and were alive 36 and 71 months after diagnosis . All cases of Ph+ AML were negative for P00519 and other gene mutations . One ( 20 % ) patient with CML-BP had P00519 mutation ; no patients had P06748 mutations . These data suggest that Ph+ AML is distinct from CML-BP . P28908 (+) anaplastic large-cell lymphoma in children : analysis of 82 patients enrolled in two consecutive studies of the French Society of Pediatric Oncology . The purpose of this study was ( 1 ) to investigate the efficacy of chemotherapy regimens designed by the French Society of Pediatric Oncology for childhood anaplastic large-cell lymphoma ( ALCL ) and ( 2 ) to identify prognostic factors in these children . Eighty-two children with newly diagnosed ALCL were enrolled in two consecutive studies , P61073 and HM91 . The diagnosis of ALCL was based on immuno-morphological features and all the cases but 2 were investigated using P37023 antibody directed to the P06748 / Q9UM73 protein associated with the 2;5 translocation . Treatment consisted of 2 courses of COPADM ( methotrexate , cyclophosphamide , doxorubicin , vincristine , and prednisone ) and a maintenance treatment of 5 to 7 months . Seventy-eight patients ( 95 % ) achieved a complete remission and 21 relapsed . The probability of survival and event-free survival at 3 years was of 83 % ( 72 % to 90 % ) and 66 % ( 54 % to 76 % ) , respectively , with a median follow-up of 49 months . In multivariate analysis , visceral involvement , mediastinal involvement , and lacticodeshydrogenase ( LDH ) level >/= 800 UI/L were shown to be predictive of a higher risk of failure . In conclusion , this type of regimen demonstrated efficacy in childhood ALCL . However , therapeutic results have to be improved for children with adverse prognostic parameters such as visceral or mediastinal involvement or a high LDH level . The O14980 nuclear export protein in normal development and disease . O14980 ( Chromosomal Maintenance 1 , also known as Exportin 1 ) is the major mammalian export protein that facilitates the transport of large macromolecules including RNA and protein across the nuclear membrane to the cytoplasm . The gene encoding O14980 was originally identified in yeast as required to maintain higher order chromosome structure . In mammalian cells , O14980 was found to bind several nuclear pore proteins hence its role in nuclear-cytosolic transport was discovered . In addition to nuclear-cytosolic transport , O14980 also plays a role in centrosome duplication and spindle assembly , especially in response to DNA damage . The crystal structure of O14980 suggests a complex protein that binds the Ran protein bound to GTP , allowing for a conformational change that facilitates binding to different cargo proteins through a nuclear export signal ( P48681 ) . Included in the cadre of cargo are multiple tumor suppressor and oncoproteins as p53 , P38398 , Survivin , P06748 , and P25054 , which function in the nucleus to regulate transcription or aid in chromosomal assembly and movement . An imbalance in the cytosolic level of these proteins has been observed in cancer cells , resulting in either inactivation ( tumor suppressor ) or an excess of anti-apoptotic activity ( oncoprotein ) . Thus , the concept of inhibiting O14980 has been explored as a potential therapeutic intervention . Indeed , inhibition of O14980 by a variety of small molecules that interfere with cargo- P48681 binding results in cancer cell death . Whether all of these proteins together are responsible for this phenotype or whether specific proteins are required for this effect is unclear at this time . Linkage analysis of multiple sclerosis with candidate region markers in Sardinian and Continental Italian families . Previous genome screens in multiple sclerosis have shown some evidence of linkage in scattered chromosomal regions . Although in no case the evidence of each single study was compelling and although in general the linkage ' peaks ' of the different studies did not coincide , some regions can be considered likely candidates for the presence of MS risk genes because of the clustering of MLS scores and homology with eae loci . We performed a linkage analysis of markers in these regions and of intragenic markers of some individual candidate genes ( HLA- Q8IUH3 , P16410 , P15248 , P02649 , P10415 , P20333 ) . For the first time , Southern European populations were targeted , namely Continental Italians and Sardinians . A total of 69 multiplex families were typed for 67 markers by a semi-automatic fluorescence-based assay . Results were analysed for linkage by two non-parametric tests : GENEHUNTER and SimIBD . In general , the linkage scores obtained were low , confirming the conclusion that no gene is playing a major role in the disease . However , some markers , in 2p11 , 3q21.1 , 7p15.2 and 22q13.1 stood out as promising since they showed higher scores with one or the other test . This stimulates further association analysis of a large number of simplex families from the same populations . Molecular targets and regulators of cardiac hypertrophy . Cardiac hypertrophy is one of the main ways in which cardiomyocytes respond to mechanical and neurohormonal stimuli . It enables myocytes to increase their work output , which improves cardiac pump function . Although cardiac hypertrophy may initially represent an adaptive response of the myocardium , ultimately , it often progresses to ventricular dilatation and heart failure which is one of the leading causes of mortality in the western world . A number of signaling modulators that influence gene expression , apoptosis , cytokine release and growth factor signaling , etc. are known to regulate heart . By using genetic and cellular models of cardiac hypertrophy it has been proved that pathological hypertrophy can be prevented or reversed . This finding has promoted an enormous drive to identify novel and specific regulators of hypertrophy . In this review , we have discussed the various molecular signal transduction pathways and the regulators of hypertrophic response which includes calcineurin , cGMP , NFAT , natriuretic peptides , histone deacetylase , P05231 cytokine family , Gq/ P49842 signaling , PI3K , MAPK pathways , Na/H exchanger , DB01367 , polypeptide growth factors , P01160 , NO , P01375 , Q07869 and JAK/ P35610 pathway , microRNA , Cardiac angiogenesis and gene mutations in adult heart . Augmented knowledge of these signaling pathways and their interactions may potentially be translated into pharmacological therapies for the treatment of various cardiac diseases that are adversely affected by hypertrophy . The purpose of this review is to provide the current knowledge about the molecular pathogenesis of cardiac hypertrophy , with special emphasis on novel researches and investigations . VIPhyb , an antagonist of vasoactive intestinal peptide receptor , enhances cellular antiviral immunity in murine cytomegalovirus infected mice . Vasoactive intestinal peptide ( P01282 ) is a neuropeptide hormone that suppresses Th1-mediated cellular immunity . We previously reported that P01282 -knockout ( P01282 -KO ) mice have enhanced cellular immune responses and increased survival following murine cytomegalovirus ( mCMV ) infection in C57BL/6 mice . In this study , we tested whether treatment with a P01282 receptor antagonistic peptide protects C57BL/6 and BALB/c mice from mCMV-infection . One week of daily subcutaneous injections of VIPhyb was non-toxic and did not alter frequencies of immune cell subsets in non-infected mice . VIPhyb administration to mCMV-infected C57BL/6 and BALB/c mice markedly enhanced survival , viral clearance , and reduced liver and lung pathology compared with saline-treated controls . The numbers of effector/memory CD8+ T-cells and mature NK cells were increased in VIPhyb-treated mice compared with PBS-treated groups . Pharmacological blockade of P01282 -receptor binding or genetic blockade of P01282 -signaling prevented the up-regulation of Q9NZQ7 and P18621 expression on DC and activated CD8+ T-cells , respectively , in mCMV-infected mice , and enhanced P33681 , P42081 , and MHC-II expression on conventional and plasmacytoid DC . VIPhyb-treatment increased type-I IFN synthesis , numbers of IFN-γ- and P01375 -α-expressing NK cells and T-cells , and the numbers of mCMV-M45 epitope-peptide-MHC-I tetramer CD8+ T-cells following mCMV infection . P01282 -treatment lowered the percentage of Treg cells in spleens compared with PBS-treated WT mice following mCMV infection , while significantly decreasing levels of serum P15692 induced by mCMV-infection . The mice in all treated groups exhibited similar levels of anti-mCMV antibody titers . Short-term administration of a P01282 -receptor antagonist represents a novel approach to enhance innate and adaptive cellular immunity in a murine model of CMV infection . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . Long-term results of a prospective randomized trial evaluating G- P04141 priming in intensive induction chemotherapy followed by autologous stem cell transplantation in elderly patients with acute myeloid leukemia . Few studies have evaluated granulocyte colony-stimulating factor ( DB00099 ) priming in elderly patients with intensively treated acute myeloid leukemia ( AML ) , and no data are available for genetically defined AML subgroups . We provide long-term results ( median follow-up 7.6 years ) of a randomized trial in which 183 patients ( median age 67 years ) received G- P04141 prior to ( G- P04141 priming ) or after two cycles of induction chemotherapy . CR rates with G- P04141 priming and G- P04141 post-chemotherapy were comparable ( 57 vs. 67 % , p = 0.153 ) , with overall survival ( OS ) probabilities of 14 vs. 17 % at 10 years . Induction mortality was significantly higher with G- P04141 priming ( 23 vs. 10 % , p = 0.015 ) , primarily in normal karyotype ( NK ) AML . In this subgroup , a trend for better relapse-free survival ( RFS ) was observed with G- P04141 priming ( 44 vs. 22 % at 10 years , p = 0.074 ) but did not translate into an OS benefit . G- P04141 priming had no impact on AML with P36888 -ITD and P06748 mutations and did not improve outcome in patients with adverse cytogenetics . In a landmark analysis , late consolidation with autologous stem cell transplantation or a second consolidation cycle significantly improved RFS compared with one consolidation cycle ( 21.0 vs. 12.8 months , p = 0.046 ) . Future studies on G- P04141 priming should be restricted to NK AML and used only in post-remission therapy . Identification of the fused bicyclic 4-amino-2-phenylpyrimidine derivatives as novel and potent DB05876 inhibitors . 2-Phenyl-4-piperidinyl-6,7-dihydrothieno[3,4-d]pyrimidine derivative ( 2 ) was found to be a new DB05876 inhibitor with moderate Q07343 activity ( IC50=150 nM ) . A number of derivatives with a variety of 4-amino substituents and fused bicyclic pyrimidines were synthesized . Among these , 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivative ( 18 ) showed potent Q07343 inhibitory activity ( IC50=25 nM ) . Finally , N-propylacetamide derivative ( 31b ) was determined as a potent inhibitor for both Q07343 ( IC50=7.5 nM ) and P01375 -α production in mouse splenocytes ( IC50=9.8 nM ) and showed good in vivo anti-inflammatory activity in the LPS-induced lung inflammation model in mice ( ID50=18 mg/kg ) . The binding mode of the new inhibitor ( 31e ) in the catalytic site of Q07343 is presented based on an X-ray crystal structure of the ligand-enzyme complex . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . P04150 interacting protein-1 restores glucocorticoid responsiveness in steroid-resistant airway structural cells . Glucocorticoid ( GC ) insensitivity represents a profound challenge in managing patients with asthma . The mutual inhibition of transcriptional activity between GC receptor ( GR ) and other regulators is one of the mechanisms contributing to GC resistance in asthma . We recently reported that interferon regulatory factor ( Q969Q1 ) -1 is a novel transcription factor that promotes GC insensitivity in human airway smooth muscle ( P17405 ) cells by interfering with GR signaling ( Tliba et al. , Am J Respir Cell Mol Biol 2008;38:463-472 ) . Here , we sought to determine whether the inhibition of GR function by P10914 involves its interaction with the transcriptional co-regulator GR-interacting protein 1 ( Q9Y3R0 ) , a known GR transcriptional co-activator . We here found that siRNA-mediated Q9Y3R0 depletion attenuated P10914 -dependent transcription of the luciferase reporter construct and the mRNA expression of an P10914 -dependent gene , P28907 . In parallel experiments , Q9Y3R0 silencing significantly reduced GR-mediated transactivation activities . Co-immunoprecipitation and Q86UG4 pull-down assays showed that Q9Y3R0 , through its repression domain , physically interacts with P10914 identifying Q9Y3R0 as a bona fide transcriptional co-activator for P10914 . Interestingly , the previously reported inhibition of GR-mediated transactivation activities by either P01375 and P01579 treatment or P10914 overexpression was fully reversed by increasing cellular levels of Q9Y3R0 . Together , these data suggest that the cellular accumulation of P10914 may represent a potential molecular mechanism mediating altered cellular response to GC through the depletion of Q9Y3R0 from the GR transcriptional regulatory complexes . Methyl gallate exhibits potent antitumor activities by inhibiting tumor infiltration of P01730 +CD25+ regulatory T cells . P01730 (+)CD25(+) regulatory T ( Treg ) cells play crucial roles in the host response to tumors . Increasing evidence supports the existence of elevated numbers of Treg cells in solid tumors and hematologic malignancies . In this study , the effects of methyl gallate on Treg cells were examined . Methyl gallate inhibited Treg cell-suppressive effects on effector P01730 (+) T cells and Treg migration toward tumor environment . The expression of Treg surface markers including P16410 , CCR4 , P61073 , and glucocorticoid-induced TNFR was significantly suppressed upon methyl gallate treatment . Furthermore , forkhead box P09131 ( Foxp3 ) expression was also significantly decreased by methyl gallate , suggesting that the suppressive effects of methyl gallate on Treg were medicated by decrease of Treg-specific transcription factor Foxp3 . In tumor-bearing hosts , methyl gallate treatment substantially reduced tumor growth and prolonged the survival rate . In contrast , nu/nu mice did not show decreased tumor progression in response to methyl gallate . In addition , in tumor-bearing Treg-depleted mice , tumor growth and the survival rates were not changed by methyl gallate treatment , strongly suggesting that the main therapeutic target of methyl gallate in tumor suppression was related to modulation of the P01730 (+)CD25(+) Treg cell functions . In the spleen of tumor-bearing mice , methyl gallate treatment induced a significant decrease in the P01730 (+)CD25(+)Foxp3(high) Treg cell population . Especially , the number of tumor-infiltrating CD25(+)Foxp3(high) Treg cells was significantly lower in methyl gallate-treated mice . These results suggest that methyl gallate can be used to reverse immune suppression and as a potentially useful adjunct for enhancing the efficacy of immune-based cancer therapy . [ Establishment of hematological malignancy model in ex vivo cell culture ] . The aim of this study was to develop an ex vivo cell culture system for establishing the hematological malignancy model . Mouse bone marrow cells were transfected with GFP-expressed retroviral vectors encoding various leukemia/lymphoma-associated fusion proteins ( P41212 - P09619 , Rabaptin5- P09619 , p210BCR- P00519 , Q01196 - Q06455 , P06748 - Q9UM73 ) . After transfection , the cells were cultured in IMDM containing 10 % FCS without growth factors , or with one of the following growth factor combinations : ( 1 ) murine c-kit ligand ( KL ) plus human flt3 ligand ( FL ) ; ( 2 ) P08700 , thrombopoietin , DB00099 , and hyper- P05231 ( 3/T/G/ Q9NP08 ) ; ( 3 ) KL/FL plus 3/T/G/ Q9NP08 . The flow cytometry was used to detect the ability of combinations of growth factors to complement the oncogene fusion protein to support self-renewal of the transfected cells . The results showed that the transfected cells could be amplified sustainably in the logarithmic growth way . The indicated combination of P21583 ( KL ) with flt-3 ligand ( FL ) supported the self-renewal of the marrow cells transfected with vectors encoding P41212 - P09619 , Rabaptin5- P09619 , Q01196 - Q06455 and P06748 - Q9UM73 , in addition to KL/FL , the self-renewal of Q92817 P11274 - P00519 transfected-marrow cells also required P08700 . The morphology of cells emerged from culture can be the predictor of the corresponding oncogene-associated malignancy . It is concluded that this study establishes a culture system ex vivo which provides a generalized method for studying hematological malignancies , and may facilitate the screening for therapeutic agents . Results of a pivotal phase II study of brentuximab vedotin for patients with relapsed or refractory Hodgkin 's lymphoma . PURPOSE : DB08870 is an antibody-drug conjugate ( ADC ) that selectively delivers monomethyl auristatin E , an antimicrotubule agent , into P28908 -expressing cells . In phase I studies , brentuximab vedotin demonstrated significant activity with a favorable safety profile in patients with relapsed or refractory P28908 -positive lymphomas . PATIENTS AND METHODS : In this multinational , open-label , phase II study , the efficacy and safety of brentuximab vedotin were evaluated in patients with relapsed or refractory Hodgkin 's lymphoma ( HL ) after autologous stem-cell transplantation ( auto- P09683 ) . Patients had histologically documented P28908 -positive HL by central pathology review . A total of 102 patients were treated with brentuximab vedotin 1.8 mg/kg by intravenous infusion every 3 weeks . In the absence of disease progression or prohibitive toxicity , patients received a maximum of 16 cycles . The primary end point was the overall objective response rate ( ORR ) determined by an independent radiology review facility . RESULTS : The ORR was 75 % with complete remission ( CR ) in 34 % of patients . The median progression-free survival time for all patients was 5.6 months , and the median duration of response for those in CR was 20.5 months . After a median observation time of more than 1.5 years , 31 patients were alive and free of documented progressive disease . The most common treatment-related adverse events were peripheral sensory neuropathy , nausea , fatigue , neutropenia , and diarrhea . CONCLUSION : The ADC brentuximab vedotin was associated with manageable toxicity and induced objective responses in 75 % of patients with relapsed or refractory HL after auto- P09683 . Durable CRs approaching 2 years were observed , supporting study in earlier lines of therapy . Donor Q9NR96 gene tagSNPs influence susceptibility to aGVHD and CMV reactivation in the allo-HSCT setting without polymorphisms in the O00206 and Q9HC29 genes . Owing to ethnicity of the population , those best confirmed polymorphisms in the TLR ( toll-like receptor ) 4 and Q9HC29 genes with significantly prognostic impact on allogeneic hematopoietic P09683 ( allo-HSCT ) seem to be more applicable to Europeans and are nonpolymorphic in the Asian population . The influence of innate immunity gene polymorphisms on the outcomes of allo-HSCT in those populations has been questioned . We evaluated the influence of 10 candidate single nucleotide polymorphisms ( SNPs ) in the Q15399 , O60603 , O15455 , Q9NR97 and Q9NR96 genes on the outcomes of allo-HSCT in a Chinese population including 138 pairs of patients and unrelated donors and a second cohort of 102 pairs of patients and HLA-identical sibling donors . We found that two tagSNPs in the Q9NR96 gene in the donor side , +1174 A/G ( rs352139 ) and +1635 C/T ( rs352140 ) , influenced the risk of acute GVHD ( aGVHD ) and CMV reactivation . Furthermore , the presence of the susceptible haplotype ( A-C ) in donor may be an informative predicator of worse OS at 5 years compared with those with the G-C and G-T haplotypes ( 58 % vs 82.9 % , P=0.024 ) . Our data suggested an unrecognized association between donor Q9NR96 tagSNPs and the risk of HSCT-related complications in a population without polymorphisms in the O00206 and Q9HC29 genes . 3,4-Dimethoxyphenyl bis-benzimidazole derivative , mitigates radiation-induced DNA damage . Radiation-induced DNA damage initiates a series of overlapping responses that include DNA damage recognition and repair , induction of cell cycle checkpoints , senescence and/or apoptosis . This study assessed the DNA damage response and whole genome expression profile in two mammalian cell lines ( P29320 and U87 ) in response to ( 5-{4-methylpiperazin-1-yl}-2-[2'-(3,4-dimethoxyphenyl)-5'-benzimidazolyl] benzimidazole ) P28067 and ionizing radiation . P28067 has been shown to act as a potent radiation protector , yielding significant levels of protection , i.e. , 20.9 % in P29320 cells and 21.2 % in U87 cells . Our findings revealed treatment with P28067 significantly reduced γ- P16104 , Q12888 and Rad51 foci formation after irradiation . Q96HU1 kinase , WNT signaling and p53 pathways were found to be activated in P28067 -treated cells . In addition , the DNA damage response genes , HSP70 , P10809 , Q06830 , Q9BXM0 , P27797 , P06748 , P0CG48 , and Q01105 showed differential regulation in P28067 , P28067 + radiation and radiation-treated cells . The data suggest that P28067 -influenced repertoire of repair proteins , which are an indispensable part of the cell , interplay with each other to reduce DNA damage and maintain the genomic integrity of the cell . [ Spiral computed tomography with functional tests in diagnosis of chronic obstructive diseases of the lungs ] . The potential of spiral computed tomography ( P09683 ) and high resolution computed tomography ( HRST ) in diagnosis of chronic obstructive pulmonary diseases ( P48444 ) is described . Semiotic sings of bronchial asthma ( BA ) and chronic obstructive bronchitis ( P00156 ) are specified . Informative value of P09683 in differential diagnosis of BA and P00156 including early stage of the diseases is analysed . An updated technique of P09683 and HRCT of the lungs in P48444 patients is presented . Inhibitor-immunology-study. Evaluation of inhibitor development in haemophilia B . The development of inhibitors in haemophilia B is one of the most important complications of replacement therapy , affecting mortality and morbidity . Inhibitor development is based on complex immunological factors , and to date , only little is known about its underlying mechanisms . Here , we present first results of the haemophilia B group of our Inhibitor-Immunology study . PATIENTS , METHODS : So far we have analysed 15 patients with haemophilia B . Four of them developed a high titre inhibitor ; the remaining 11 had no inhibitor . We evaluated 9 SNPs in 8 genes ( P25942 , P16410 , IL-1β , P22301 , O60603 , O00206 , Q9NR96 , P01375 -α ) . We compared the distribution of these alleles between inhibitor and non-inhibitor haemophilia B patients and between haemophilia B patients and a normal male control population . HLA typing was performed in all patients . Results , discussion : There appears to be a trend towards a skewed distribution of TLR 9 , P22301 and P16410 alleles in haemophilia B patients . Due to the limited number these differences are , however , not statistically significant . The t-test of all patients with inhibitor versus without inhibitor was significant for HLA-A*03 and DPB1*0401 and borderline for Q8IUH3 *0201 . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Association between high interleukin-6 levels and adverse outcome after autologous haemopoietic stem cell transplantation . We studied interleukin-6 ( P05231 ) levels on the day of transplantation in 31 patients undergoing autologous haemopoietic stem cell transplantation ( P09683 ) ( either peripheral blood stem cell transplantation ( PBSCT ) or bone marrow transplantation ( BMT ) ) for neoplastic diseases to determine if there was a relationship between P05231 level and rate of haemopoietic recovery , length of stay in hospital , and survival . There was no apparent delay in post-transplant recovery associated with elevated P05231 levels . However , increased values of P05231 tended to be associated with an increased length of stay in hospital ( P = 0.083 ) . There was a highly significant adverse association between higher P05231 levels and survival following transplantation ( P = 0.0001 ) . This association remained significant ( P = 0.013 ) in the uniform subgroup of patients with malignant lymphoma with chemosensitive disease who had undergone BMT ( that is , excluding patients who had undergone PBSCT ) ( n = 13 ) . Knowledge of P05231 levels on the day of transplant has the potential to provide valuable prognostic information in patients undergoing autologous haemopoietic P09683 . Thalidomide suppresses Up-regulation of human immunodeficiency virus coreceptors P61073 and P51681 on P01730 + T cells in humans . Concurrent infection in patients with human immunodeficiency virus ( HIV ) infection increases the expression of HIV coreceptors P61073 and P51681 . Thalidomide has beneficial effects in a number of HIV-associated diseases . The effect of thalidomide on P61073 and P51681 expression on P01730 + T cells was determined . Thalidomide produced a dose-dependent inhibition of lipopolysaccharide ( LPS ) -induced up-regulation of P61073 and P51681 in vitro . Antibody to tumor necrosis factor-alpha ( P01375 ) also attenuated the LPS-induced HIV coreceptor up-regulation , which was not further reduced by thalidomide . Thalidomide ( 400 mg ) was orally administered to 6 men , and their blood was stimulated ex vivo with LPS , staphylococcal or mycobacterial antigens , or antibody to CD3 or P10747 cells . All stimuli induced up-regulation of HIV coreceptors , which was reduced after ingestion of thalidomide . Thalidomide may be beneficial in the treatment of intercurrent infections during HIV infection by reducing the up-regulation of P61073 and P51681 expression on P01730 + T cells induced by bacterial and mycobacterial antigens , by a mechanism that involves inhibition of P01375 . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . P16410 polymorphisms and P48444 . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Genetic determinants of diabetes are similarly associated with other immune-mediated diseases . PURPOSE OF REVIEW : I discuss the increasing number of genes discovered to exert pleiotropic action on susceptibility to diabetes and simultaneously to other immune-mediated diseases . While a genetic correlation between type 1 diabetes and autoimmunity is not surprising , the mechanism of a relationship to other conditions such as atopy is less obvious . The recent wave of genome-wide association studies offers confirmation of previously recognized risk loci , but also novel loci that also are likely to act via multiple pathogenetic pathways . I will review this material more in a genetical than an immunological way . RECENT FINDINGS : Identification of Q9BYX4 , an RNA helicase involved in the innate immune response to viral infection as a risk factor for type 1 diabetes and rheumatoid arthritis . Confirmation of the roles of P16410 and Q9Y2R2 as general immune function modulators with a nonlinear dose-response effect on autoimmunity , and confirmation of the role of P01589 , which may act via a regulatory T cell subset on immune disease risk . Expansion of the role of P37231 in immunity . SUMMARY : The wave of genome-wide association studies already experienced in the last 2 years has solidified what we thought we knew and added a list of genes in new pathways . The association of Q9BYX4 with type 1 diabetes may offer a real window into early events in the development of that disease . DB08870 : its role in the treatment of anaplastic large cell and Hodgkin 's lymphoma . DB08870 is being developed in a joint collaboration between Seattle Genetics and Millennium : The Takeda Oncology Company . In August 2011 , it was approved by the FDA for the treatment of patients with Hodgkin 's lymphoma ( HL ) and anaplastic large cell lymphoma ( ALCL ) . DB08870 is an antibody-drug conjugate that specifically targets the P01375 receptor superfamily member 8 ( P28908 ) antigen on the surface of cancer cells to induce cell death . DB08870 has shown efficacy in inducing apoptosis in HL and ALCL cell lines that express P28908 and reducing tumor size in preclinical models . DB08870 is under clinical evaluation for the treatment of relapsed or refractory HL and ALCL in both adults and children . It is being investigated for use as a combination agent with pre-existing frontline chemotherapies and as a stand-alone salvage therapy for use prior to autologous stem cell transplant . Treatment with brentuximab vedotin is generally well tolerated although it is associated with grade 1-2 adverse reactions such as neutropenia and there have been reports of grade 3-4 serious adverse events . In particular its use with chemotherapy regimens that include bleomycin is contraindicated because of adverse pulmonary effects . Role for macrophage migration inhibitory factor in acute respiratory distress syndrome . The critical role of macrophage migration inhibitory factor ( MIF ) in mediating inflammatory lung injury in acute respiratory distress syndrome ( ARDS ) has been raised recently . The present study has identified enhanced MIF protein expression in alveolar capillary endothelium and infiltrating macrophages in lung tissues from ARDS patients . The possibility that MIF up-regulates its synthesis in an autocrine fashion in ARDS was tested using cultured endothelial cells stimulated with MIF and a murine model of lipopolysaccharide ( LPS ) -induced acute lung injury . MIF induced significant MIF and tumour necrosis factor ( P01375 ) -alpha synthesis in cultured endothelial cells and the effect was blocked by neutralizing anti-MIF antibody . A similar blocking effect was observed when MIF-stimulated endothelial cells were pretreated with neutralizing anti- P01375 antibody or glucocorticoid , supporting the notion that MIF induced P01375 production via an amplifying pro-inflammatory loop . Treatment with anti-MIF or glucocorticoid effectively attenuated pulmonary pathology and the synthesis of MIF or P01375 in mice with LPS-induced acute lung injury . Mildly augmented expression of aquaporin 1 ( P29972 ) was also detected in alveolar capillary endothelium in ARDS . In vitro studies revealed that both MIF and P01375 induced a small increase of P29972 synthesis in cultured endothelial cells . These findings suggest that MIF plays a crucial pathological role leading to alveolar inflammation in ARDS . Anti-MIF and early glucocorticoid therapy may represent a novel therapeutic approach for reducing alveolar inflammation in ARDS . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . DB09037 . The development of the cytotoxic T-lymphocyte-associated protein 4 inhibitor ipilimumab and its approval in 2011 for the treatment of metastatic melanoma has heralded a new era in immuno-oncology . Subsequently , novel agents against the programmed death receptor 1 ( P18621 ) /programmed death receptor ligand 1 ( Q9NZQ7 ) axis have shown significant activity in melanoma and a variety of other tumor types . DB09037 was the first anti- P18621 antibody to be approved by the US Food and Drug Administration for the treatment of patients with unresectable or metastatic melanoma with disease progression following ipilimumab , and if P15056 ( V600 ) mutation positive , a P15056 inhibitor . DB09037 has also received breakthrough status for the treatment of P00533 mutation-negative , Q9UM73 rearrangement-negative non-small cell lung cancer ( NSCLC ) that has progressed on or following platinum-based chemotherapy . There remain a number of pivotal trials in progress to further evaluate the optimal use of pembrolizumab alone and in combination for melanoma , NSCLC , and other tumor types . In this article , we review the efficacy and toxicity profile of pembrolizumab and evaluate its future development . Understanding factors that inhibit or promote the utilization of telecare in chronic lung disease . OBJECTIVES : To perform a process evaluation of a randomized controlled trial ( RCT ) of home telecare for the management of acute exacerbations of chronic obstructive pulmonary disease ( P48444 ) , using the normalization process model ( P06748 ) as an explanatory framework . METHODS : Semi-structured interviews were carried out with patients ( n = 9 ) and nurses ( n = 11 ) participating in a RCT . A framework approach to data analysis was used . RESULTS : The telecare service did not provide an interactional advantage for the nurses providing this service and did not fit with the nurses ' views of the most appropriate or preferred use of their skills . The telecare service seemed unlikely to become normalized as part of routine healthcare delivery , because the nursing team lacked confidence that it was a safe way to provide healthcare in this context and it was not perceived as improving efficiency . DISCUSSION : The P06748 effectively mapped onto the study findings and explained those factors that inhibited the routine delivery of P48444 services by telecare . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . P42345 signaling regulates nucleolar targeting of the SUMO-specific isopeptidase Q9H4L4 . Ribosome biogenesis is a multistep cellular pathway that involves more than 200 regulatory components to ultimately generate translation-competent 80S ribosomes . The initial steps of this process , particularly rRNA processing , take place in the nucleolus , while later stages occur in the nucleoplasm and cytoplasm . One critical factor of 28S rRNA maturation is the SUMO-isopeptidase Q9H4L4 . Q9H4L4 tightly interacts with the nucleolar scaffold protein P06748 and is associated with nucleolar 60S preribosomes . A central question is how changes in energy supply feed into the regulation of ribosome maturation . Here , we show that the nutrient-sensing P42345 kinase pathway controls the nucleolar targeting of Q9H4L4 by regulating its interaction with P06748 . We define an N-terminal domain in Q9H4L4 as the critical P06748 binding region and provide evidence that P42345 -mediated phosphorylation of serine/threonine residues within this region fosters the interaction of Q9H4L4 with P06748 . The inhibition of P42345 triggers the nucleolar release of Q9H4L4 , thereby likely compromising its activity in rRNA processing . Since P42345 activity is tightly coupled to nutrient availability , we propose that this pathway contributes to the adaptation of ribosome maturation in response to the cellular energy status . Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells . Antibody-drug conjugates ( ADC ) are emerging as powerful treatment strategies with outstanding target-specificity and high therapeutic activity in patients with cancer . DB08870 represents a first-in-class ADC directed against P28908 (+) malignancies . We hypothesized that its sustained clinical responses could be related to the stimulation of an anticancer immune response . In this study , we demonstrate that the dolastatin family of microtubule inhibitors , from which the cytotoxic component of brentuximab vedotin is derived , comprises potent inducers of phenotypic and functional dendritic cell ( DC ) maturation . In addition to the direct cytotoxic effect on tumor cells , dolastatins efficiently promoted antigen uptake and migration of tumor-resident DCs to the tumor-draining lymph nodes . Exposure of murine and human DCs to dolastatins significantly increased their capacity to prime T cells . Underlining the requirement of an intact host immune system for the full therapeutic benefit of dolastatins , the antitumor effect was far less pronounced in immunocompromised mice . We observed substantial therapeutic synergies when combining dolastatins with tumor antigen-specific vaccination or blockade of the P18621 - Q9NZQ7 and P16410 coinhibitory pathways . Ultimately , treatment with ADCs using dolastatins induces DC homing and activates cellular antitumor immune responses in patients . Our data reveal a novel mechanism of action for dolastatins and provide a strong rationale for clinical treatment regimens combining dolastatin-based therapies , such as brentuximab vedotin , with immune-based therapies . DB08870 in anaplastic large cell lymphoma . INTRODUCTION : DB08870 , a novel anti- P28908 antibody-drug conjugate , delivers a cytotoxic agent into P28908 (+) cells . P28908 expression is characteristic of anaplastic large cell lymphoma ( ALCL ) and Hodgkin lymphoma ( HL ) . AREAS COVERED : We reviewed data on brentuximab vedotin , focusing on ALCL and discuss pharmacology , clinical trials leading to approval and future research directions . Systemic ALCL , 3 % of adult Q9NZ71 , is characterized by large anaplastic P28908 (+) cells . The fusion protein P06748 - Q9UM73 , when present in systemic ALCL , confers better prognosis , although even Q9UM73 - patients with IPI score ≥ 3 are high-risk . For patients with systemic ALCL , 25 - 45 % relapse after frontline therapy , and > 50 % of patients will relapse following high-dose chemotherapy with autologous stem-cell support . There has been no standard therapy for relapsed/refractory systemic ALCL . DB08870 , combines a monoclonal antibody targeted to P28908 with a microtubule disrupting agent and was recently approved for treatment of patients with systemic ALCL that is refractory or relapsed after at least one multiagent chemotherapy regimen . EXPERT OPINION : DB08870 provides targeted therapy to P28908 (+) lymphomas , including ALCL and HL , with high response rates and manageable toxicity , predominantly myelosuppression and peripheral neuropathy . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Mutation of beta-catenin and its protein accumulation in solid and cystic tumor of the pancreas associated with metastasis . P35222 serves not only as a structural component of the P12830 -mediated cell-cell adhesion system , but also as a signaling molecule of the Wnt/wingless pathway . Mutations of beta-catenin and aberrant expression of its protein have been identified in a number of different types of human malignancies . To determine the role of beta-catenin in solid and cystic tumor ( P09683 ) of the pancreas , a rare neoplasm usually observed in young females , we examined three primary tumors and one corresponding liver metastatic tumor presented in pediatric patients . Single strand conformation polymorphism and neucleotide sequencing analysis confirmed a TCT right curved arrow TTT somatic mutation at codon 37 changing serine to phenylalanine in one of the primary tumors and its corresponding metastatic tumor . Immunohistochemical analysis displayed abnormally strong nuclear and widespread cytoplasmic expression of beta-catenin in these tumors with the mutation . Our observations suggest that , in some SCTs of the pancreas presented in the pediatric age group , mutated beta-catenin has an important oncogenic effect . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Cordycepin suppresses P01375 -α-induced NF-κB activation by reducing p65 transcriptional activity , inhibiting IκBα phosphorylation , and blocking IKKγ ubiquitination . Cordycepin is reported to participate in multiple pharmacological activities including anti-tumor and anti-inflammation , and is involved in the regulation of NF-κB signaling pathway . However , the detailed molecular mechanism of cordycepin in suppression of NF-κB signaling pathway remains ambiguous . In this study , we first analyzed the effect of cordycepin on NF-κB activity in P29320 -293T cells , and found that cordycepin resulted in a dose-dependent reduction in P01375 -α-induced NF-κB activation . Although cordycepin did not block P01375 -α-induced nuclear translocation of p65 , high concentration of cordycepin reduced the DNA-binding and transcriptional activities of NF-κB . Moreover , cordycepin also inhibited IκBα phosphorylation so as to suppress the degradation of IκBα . Further investigation revealed that cordycepin suppressed IKKs-mediated NF-κB activation and inhibited the ubiquitination of IKKγ . In conclusion , cordycepin effectively inhibits NF-κB signaling through suppressing the activities of NF-κB , IκB and IKK . Thus , cordycepin may provide some potential therapeutic application in inflammation-associated disorders and cancer . Efficient mobilization of PBSC with vinorelbine/G- P04141 in patients with malignant lymphoma . High-dose chemotherapy ( HDT ) and hematopoietic P09683 are effective in patients with relapsing or refractory malignant lymphoma . Collection of sufficient numbers of stem cells is a prerequisite for such a therapy . In a pilot trial , we evaluated the feasibility of stem cell mobilization with vinorelbine/G- P04141 in patients with lymphoma , a regimen allowing precise timing and harvesting of sufficient stem cells in myeloma patients . Forty-five patients with lymphoma received vinorelbine 35 mg/m(2) i.v. on day 1 and G- P04141 10 microg/kg/day s.c. , divided in two daily doses from day 4 until collection . Stem cell collection was successfully performed in 43 patients ( 96 % ) with a median of 3.6 x 10(6) P28906 (+) cells/kg ( range : 1.4-16 ) in the collected product . In 28 patients ( 62 % ) , the first stem cell apheresis was performed on day 8 , and for 28 patients a sufficient stem cell yield was reached with one apheresis only . All 43 patients underwent high-dose chemotherapy with BEAM and auto- P09683 with hematological recovery on time and without unexpected toxicity . In conclusion , vinorelbine/G- P04141 allows accurate timing and safe harvesting of sufficient stem cells in patients with malignant lymphoma . P62158 interacts with angiotensin-converting enzyme-2 ( Q9BYF1 ) and inhibits shedding of its ectodomain . P12821 -2 ( Q9BYF1 ) is a regulatory protein of the renin-angiotensin system ( DB01367 ) and a receptor for the causative agent of severe-acute respiratory syndrome ( P49591 ) , the P49591 -coronavirus . We have previously shown that Q9BYF1 can be shed from the cell surface in response to phorbol esters by a process involving P01375 converting enzyme ( P78536 ; P78536 ) . In this study , we demonstrate that inhibitors of calmodulin also stimulate shedding of the Q9BYF1 ectodomain , a process at least partially mediated by a metalloproteinase . We also show that calmodulin associates with Q9BYF1 and that this interaction is decreased by calmodulin inhibitors . The transcriptional coactivator DRIP/mediator complex is involved in vitamin D receptor function and regulates keratinocyte proliferation and differentiation . Mediator is a multisubunit coactivator complex that facilitates transcription of nuclear receptors . We investigated the role of the mediator complex as a coactivator for vitamin D receptor ( P11473 ) in keratinocytes . Using P11473 affinity beads , the vitamin D receptor interacting protein ( DRIP ) /mediator complex was purified from primary keratinocytes , and its subunit composition was determined by mass spectrometry . The complex included core subunits , such as Q15648 /MED1 ( MED1 ) , that directly binds to P11473 . Additional subunits were identified that are components of the RNA polymerase II complex . The functions of different mediator components were investigated by silencing its subunits . The core subunit MED1 facilitates P11473 activity and regulating keratinocyte proliferation and differentiation . A newly described subunit Q13503 also has a role in promoting keratinocyte proliferation and differentiation , whereas Q9BTT4 has an inhibitory role . Blocking MED1/ Q13503 expression caused hyperproliferation of keratinocytes , accompanied by increases in mRNA expression of the cell cycle regulator cyclin D1 and/or glioma-associated oncogene homolog . Blocking MED1 or Q13503 expression also resulted in defects in calcium-induced keratinocyte differentiation , as indicated by decreased expression of differentiation markers and decreased translocation of P12830 to the membrane . These results show that keratinocytes use the transcriptional coactivator mediator to regulate P11473 functions and control keratinocyte proliferation and differentiation .

MH_train_45

MH_train_45

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interacts_with DB01278?

multiple_choice

Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Renal medullary carcinoma : clinical , pathologic , immunohistochemical , and genetic analysis with pathogenetic implications . OBJECTIVES : To investigate the pathologic , clinical , and genetic features of renal medullary carcinomas ( RMCs ) in search of clues to their pathogenesis . METHODS : We analyzed 40 RMCs for clinical features , for immunohistochemical expression using a panel of markers , and for genetic changes using comparative genomic hybridization . RESULTS : Patients presented at 5 to 32 years of age , and 82 % were African American . All patients tested had sickle cell trait or disease . Seven patients presented with suspected renal abscess or urinary track infection without a clinically recognizable mass . Of the 15 tumors able to be analyzed , all were positive for epithelial markers P62158 5.2 and epithelial membrane antigen . All were negative for high-molecular-weight cytokeratin 34betaE12 . Cytokeratins 7 and 20 and carcinoembryonic antigen were heterogeneous and variable . Ulex was focally positive in a minority of cases . Eight of 12 tumors showed significant positivity for P04637 protein ( greater than 25 % nuclear positivity ) . All tumor tested ( n = 8 ) were strongly positive for vascular endothelial growth factor and hypoxia inducible factor . Of nine tumors analyzed for genetic gains and losses using comparative genomic hybridization , eight showed no changes and one showed loss of chromosome 22 . Survival ranged from 2 weeks to 15 months ( mean 4 months ) . CONCLUSIONS : These findings suggest that RMC is clinically and pathologically distinct from collecting duct carcinoma . The hypothesis that chronic medullary hypoxia secondary to hemoglobinopathy may be involved in the pathogenesis of RMC is suggested by strong vascular endothelial growth factor and hypoxia inducible factor expression and positivity for P04637 . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . P10997 -driven metabolic reprogramming induces regression of p53-deficient tumours in vivo . P04637 is commonly altered in human cancer , and Tp53 reactivation suppresses tumours in vivo in mice ( P04637 and Tp53 are also known as p53 ) . This strategy has proven difficult to implement therapeutically , and here we examine an alternative strategy by manipulating the p53 family members , Tp63 and Tp73 ( also known as p63 and p73 , respectively ) . The acidic transactivation-domain-bearing ( TA ) isoforms of p63 and p73 structurally and functionally resemble p53 , whereas the ΔN isoforms ( lacking the acidic transactivation domain ) of p63 and p73 are frequently overexpressed in cancer and act primarily in a dominant-negative fashion against p53 , TAp63 and TAp73 to inhibit their tumour-suppressive functions . The p53 family interacts extensively in cellular processes that promote tumour suppression , such as apoptosis and autophagy , thus a clear understanding of this interplay in cancer is needed to treat tumours with alterations in the p53 pathway . Here we show that deletion of the ΔN isoforms of p63 or p73 leads to metabolic reprogramming and regression of p53-deficient tumours through upregulation of P10997 , the gene that encodes amylin , a 37-amino-acid peptide co-secreted with insulin by the β cells of the pancreas . We found that P10997 is causally involved in this tumour regression and that amylin functions through the calcitonin receptor ( CalcR ) and receptor activity modifying protein 3 ( O60896 ) to inhibit glycolysis and induce reactive oxygen species and apoptosis . DB01278 , a synthetic analogue of amylin that is currently used to treat type 1 and type 2 diabetes , caused rapid tumour regression in p53-deficient thymic lymphomas , representing a novel strategy to target p53-deficient cancers . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Antipsychotic-like actions of the satiety peptide , amylin , in ventral striatal regions marked by overlapping calcitonin receptor and RAMP-1 gene expression . P10997 is a calcitonin-related peptide co-secreted with insulin , which produces satiety through brainstem-localized receptors ; however , its effects in forebrain are poorly understood . The nucleus accumbens shell ( AcbSh ) exhibits among the densest concentrations of high-affinity amylin binding ; nevertheless , these receptors have not been explored beyond one study showing dopamine antagonist-like effects of intra-Acb amylin on feeding and associated behavior ( Baldo and Kelley , 2001 ) . Here , we investigated whether intra-Acb amylin signaling modulates prepulse inhibition ( PPI ) , a measure of sensorimotor gating deficient in several illnesses including schizophrenia . First , in situ hybridization revealed marked anatomical gradients for both receptor activity-modifying protein-1 ( RAMP-1 ) and calcitonin receptor gene ( P30988 ) expression in striatum [ coexpression of these genes yields a high-affinity amylin-1 receptor ( AMY1-R ) ] , with highest overlap in the medial AcbSh . Intra-AcbSh amylin infusions in rats ( 0 , 30 , and 100 ng ) reversed amphetamine ( P49418 ) -induced PPI disruption without affecting baseline startle ; dorsal striatal amylin infusions had no effect . Coinfusion of AC187 ( 20 μg ) , an antagonist for AMY1-R , blocked the ability of amylin to normalize P49418 -induced PPI disruption , showing the specificity of AcbSh amylin effects to the AMY1-R . Intra-AcbSh AC187 on its own disrupted PPI in a haloperidol-reversible manner ( 0.1 mg/kg ) . Thus , AMY1-R may be a potential target for the development of putative antipsychotics or adjunct treatments that oppose metabolic side effects of current medications . Moreover , AMY1-Rs may represent a novel way to modulate activity preferentially in ventral versus dorsal striatum . Dopamine modulating drugs influence striatal (+)-[11C]DTBZ binding in rats : Q05940 binding is sensitive to changes in vesicular dopamine concentration . Binding of (+)-[11C]DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of (+)-[11C]DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , +14 % ) or DB01576 ( d- P49418 , 20 mg/kg , +12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , -16 % ) or levodopa ( -20 % ) . We suggest that in vivo (+)-[11C]DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed . Discovery and structure-activity relationship of ( 1R ) -8-chloro-2,3,4,5-tetrahydro-1-methyl-1H-3-benzazepine ( DB04871 ) , a selective serotonin P28335 receptor agonist for the treatment of obesity . The synthesis and SAR of a novel 3-benzazepine series of P28335 agonists is described . Compound 7d ( lorcaserin , APD356 ) was identified as one of the more potent and selective compounds in vitro ( pEC50 values in functional assays measuring [(3)H]phosphoinositol turnover : P28335 = 8.1 ; 5- Q13049 = 6.8 ; P41595 = 6.1 ) and was potent in an acute in vivo rat food intake model upon oral administration ( ED50 at 6 h = 18 mg/kg ) . DB04871 was further characterized in a single-dose pharmacokinetic study in rat ( t1/2 = 3.7 h ; F = 86 % ) and a 28-day model of weight gain in growing Sprague-Dawley rat ( 8.5 % decrease in weight gain observed at 36 mg/kg b.i.d. ) . DB04871 was selected for further evaluation in clinical trials for the treatment of obesity . Molecular mechanisms of low dose ionizing radiation-induced hormesis , adaptive responses , radioresistance , bystander effects , and genomic instability . PURPOSES : To review research progress on the molecular mechanisms of low dose ionizing radiation ( LDIR ) -induced hormesis , adaptive responses , radioresistance , bystander effects , and genomic instability in order to provide clues for therapeutic approaches to enhance biopositive effects ( defined as radiation-induced beneficial effects to the organism ) , and control bionegative effects ( defined as radiation-induced harmful effects to the organism ) and related human diseases . CONCLUSIONS : Experimental studies have indicated that Ataxia telangiectasia-mutated ( Q13315 ) , extracellular signal-related kinase ( P29323 ) , mitogen-activated protein kinase ( MAPK ) , phospho-c-Jun NH(2)-terminal kinase ( JNK ) and protein 53 ( P04637 ) -related signal transduction pathways may be involved in LDIR-induced hormesis ; MAPK , P04637 may be important for adaptive response ; Q13315 , cyclooxygenase-2 ( P35354 ) , P29323 , JNK , reactive oxygen species ( ROS ) , P04637 for radioresistance ; P35354 , P29323 , MAPK , ROS , tumor necrosis factor receptor alpha ( TNFα ) for LDIR-induced bystander effect ; whereas Q13315 , P29323 , MAPK , P04637 , ROS , TNFα-related signal transduction pathways are involved in LDIR-induced genomic instability . These results suggest that different manifestations of LDIR-induced cellular responses may have different signal transduction pathways . On the other hand , LDIR-induced different responses may also share the same signal transduction pathways . For instance , P04637 has been involved in LDIR-induced hormesis , adaptive response , radioresistance and genomic instability . Current data therefore suggest that caution should be taken when designing therapeutic approaches using LDIR to induce beneficial effects in humans . Systematic meta-analyses and field synopsis of genetic association studies in schizophrenia : the SzGene database . In an effort to pinpoint potential genetic risk factors for schizophrenia , research groups worldwide have published over 1,000 genetic association studies with largely inconsistent results . To facilitate the interpretation of these findings , we have created a regularly updated online database of all published genetic association studies for schizophrenia ( ' SzGene ' ) . For all polymorphisms having genotype data available in at least four independent case-control samples , we systematically carried out random-effects meta-analyses using allelic contrasts . Across 118 meta-analyses , a total of 24 genetic variants in 16 different genes ( P02649 , P21964 , DAO , P21728 , P14416 , P21917 , Q96EV8 , P47870 , Q13224 , HP , P01584 , P42898 , O75051 , P31645 , P04637 and P17752 ) showed nominally significant effects with average summary odds ratios of approximately 1.23 . Seven of these variants had not been previously meta-analyzed . According to recently proposed criteria for the assessment of cumulative evidence in genetic association studies , four of the significant results can be characterized as showing ' strong ' epidemiological credibility . Our project represents the first comprehensive online resource for systematically synthesized and graded evidence of genetic association studies in schizophrenia . As such , it could serve as a model for field synopses of genetic associations in other common and genetically complex disorders . E3 ubiquitin ligase Q13049 negatively regulates tumor suppressor p53 to promote tumorigenesis . P04637 has a key role in maintaining genomic stability and preventing tumorigenesis through its regulation of cellular stress responses , including apoptosis , cell cycle arrest and senescence . To ensure its proper levels and functions in cells , p53 is tightly regulated mainly through post-translational modifications , such as ubiquitination . Here , we identified E3 ubiquitin ligase Q13049 as a novel p53 target gene and negative regulator to regulate p53-mediated stress responses . In response to stress , such as DNA damage , p53 binds to the p53 responsive element in the promoter of the Q13049 gene and transcriptionally induces the expression of Q13049 in cells . In turn , Q13049 interacts with p53 and promotes p53 degradation through ubiquitination . Thus , Q13049 negatively regulates p53-mediated apoptosis , cell cycle arrest and senescence in response to stress . Q13049 is frequently overexpressed in different types of human tumors . Q13049 overexpression promotes cell oncogenic transformation and tumorigenesis in mice in a largely p53-dependent manner . Taken together , our results demonstrated that as a novel p53 target and a novel negative regulator for p53 , Q13049 has an important role in regulation of p53 and p53-mediated cellular stress responses . Furthermore , our results also revealed that impairing p53 function is a novel mechanism for Q13049 in tumorigenesis . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Biomarkers for assessing therapeutic response in bladder cancer . Reliable markers for assessing therapeutic response are needed to select the most effective treatment strategy for bladder cancer patients . We analyzed the role of biomarkers predicting response of non-muscle invasive bladder cancer ( NMIBC ) on BCG induction , and of non-organ confined muscle invasive bladder cancer ( MIBC ) on neoadjuvant chemotherapy . A critical , non-structured review of the literature was conducted . For assessing BCG therapy outcome , measurement of urinary P60568 levels seems to be the most potent marker of all the clinical parameters reviewed . Measurements of urinary interleukins P10145 , Q14116 , and tumour necrosis factor apoptosis-inducing ligand levels seem promising as well . Immunohistochemical markers ( ie , P04637 , Ki-67 , and Rb ) display contradictory results and seem unsuitable . Gene polymorphisms need to be studied more thoroughly before their clinical relevance can be determined . Regarding assessing and predicting response of MIBC to neoadjuvant chemotherapy , a set of potent markers has been studied . However , no conclusive evidence is yet available on their additional value over the established clinicopathological variables . Prospective trials are needed to validate the clinical benefit of molecular markers to predict response to BCG ( NMIBC ) and neoadjuvant chemotherapy ( MIBC ) before predictive biomarkers can become part of clinical practice . Determination of ancestral allele for possible human cancer-associated polymorphisms . To determine ancestral allele in possible cancer-associated polymorphisms , DNA samples from 10 chimpanzees ( Pan troglodytes ) were sequenced for alleles corresponding to 17 polymorphisms : 8 short tandem repeats [ P18510 ( alias IL-1RA ) variable number tandem repeat ( VNTR ) ; P04818 ( previously TS ) VNTR ; AR CAG repeat ; dinucleotide repeats of P22309 , IGF1 , P01579 ( alias P01579 ) , P03372 ( alias P03372 ) , and P00533 ] and 9 single nucleotide polymorphisms ( P03956 -1607 1G/2G , P08254 -1171 5A/6A , O15527 Ser326Cys , P05091 Gly487Lys , P04637 Arg72Pro , Q9UNQ0 Gln141Lys , P16455 Leu84Phe , P04179 Ala-9Val , and P42898 Ala222Val ) . No chimpanzee polymorphism corresponded to human P18510 VNTR ; the ancestral allele was a repeat lost in humans . Dinucleotide repeat polymorphisms of IGF1 , P01579 , P03372 , and P00533 were shared by chimpanzees , but the length of repeats tended to be longer in humans than in chimpanzees . This tendency was particularly evident for IGF1 . All of the SNPs tested are human-specific nucleotide changes . The ancestral allele 7A was shown to be lost in P08254 -1171 5A/6A . Thus , all of the possible cancer-associated polymorphisms tested have human-specific alleles , and the ancestral allele is lost in three polymorphisms ( P18510 VNTR , P22309 CA repeat , and P08254 -1171 5A/6A ) , suggesting a possible involvement of human-specific alleles in cancer susceptibility . Variance in the expression of DB00544 pathway genes in colorectal cancer . Although colorectal cancer has the third highest cancer mortality rate , the treatment remains far from optimized with patients showing variable responses to standard treatment . Molecular differences in pharmacologically relevant genes may contribute to the variability in response . This study used Taqman PCR to investigate the expression of 24 5-fluorouracil ( DB00544 ) pathway genes in colorectal cancer using paired nontumor and tumor sample from 52 patients with Dukes ' C colon cancer . In comparing tumor versus nonmalignant tissue , 14 of the 24 genes showed significant variation in gene expression . For 11 of these same genes ( Q05932 , P00374 , Q92820 , P15531 , P22392 , P31350 , Q8TCD5 , P13051 , P11172 , P04637 , and P04183 ) , a significant proportion of the patients showed an over expression of the particular gene in tumor tissue with a tumor-to-nonmalignant ( T/N ) ratio > 1.2 , whereas one gene ( Q12882 ) showed the converse with a large number of patients showing a lower expression in the tumor tissue ( T/N < 0.8 ) . Multiple gene correlations for the genes of the DB00544 pathway were found with the Spearman rank correlation of > 0.6 ( all P > 0.001 ) , suggesting possible coregulation mechanisms . Hierarchical clustering analysis created at least three groups of genes , which were consistent with groupings by the other statistical methods . Additionally , the hierarchical clustering showed two distinct groups of patients based on their gene expression . These variations in gene expression could provide valuable insights for optimizing treatment selection for patients with colorectal cancer . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers .

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[ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . P01308 resistance is not exhibited by advanced chronic obstructive pulmonary disease patients . We have previously reported increased blood glucose concentrations and skeletal muscle glycogen depletion in severe P48444 patients with chronic respiratory failure . In order to see if insulin resistance exists in severe P48444 , we investigated nine patients with advanced P48444 with chronic hypoxaemia and seven healthy control subjects of similar age , using the euglycaemic hyperinsulinaemic glucose clamp technique . We could not demonstrate a subnormal intravenous glucose requirement in response to insulin when maintaining euglycaemia in the P48444 patients with chronic hypoxaemia . This indicates that the net metabolism of glucose in P48444 patients with chronic hypoxaemia is not resistant to insulin . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . P00747 activator inhibitor-1 promotes inflammatory process induced by cigarette smoke extraction or lipopolysaccharides in alveolar epithelial cells . P00747 activator inhibitor-1 ( P05121 ) plays a role in regulating levels of some cytokines and cell migration in addition to its classic role in inhibiting fibrinolysis . P05121 levels in induced-sputum of chronic obstructive pulmonary disease ( P48444 ) patients were elevated significantly and correlated negatively with pulmonary function . To study the role of P05121 in inflammatory process in P48444 , the authors transfected alveolar epithelial cells ( AECs ) with siRNA-targeted P05121 and stimulated the cells by cigarette smoke extraction ( CSE ) or lipopolysaccharides ( LPS ) . The expression of inflammatory factors , interleukin-8 ( P10145 ) and leukotriene B4 ( LTB4 ) , and the monocyte migration were detected . The exposure to CSE or LPS induced the expression of P05121 , P10145 , and LTB4 in AECs and monocyte migration to AECs . However , they were attenuated after transfection with siRNA-targeted P05121 . In conclusion , P05121 stimulates inflammation induced by CSE or LPS in AECs through up-regulation of inflammatory factors and promoting inflammatory cell migration . P05121 may play a proinflammatory role in the pathogenesis of P48444 . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Targeting the epigenome in the treatment of asthma and chronic obstructive pulmonary disease . Epigenetic modification of gene expression by methylation of DNA and various post-translational modifications of histones may affect the expression of multiple inflammatory genes . Acetylation of histones by histone acetyltransferases activates inflammatory genes , whereas histone deacetylation results in inflammatory gene repression . Corticosteroids exert their antiinflammatory effects partly by inducing acetylation of antiinflammatory genes , but mainly by recruiting histone deacetylase-2 ( Q92769 ) to activated inflammatory genes . Q92769 deacetylates acetylated glucocorticoid receptors so that they can suppress activated inflammatory genes in asthma . In chronic obstructive pulmonary disease ( P48444 ) , there is resistance to the antiinflammatory actions of corticosteroids , which is explained by reduced activity and expression of Q92769 . This can be reversed by a plasmid vector , which restores Q92769 levels , but may also be achieved by low concentrations of theophylline . Oxidative stress causes corticosteroid resistance by reducing Q92769 activity and expression by activation of phosphoinositide-3-kinase-delta , resulting in Q92769 phosphorylation via a cascade of kinases . DB00277 reverses corticosteroid resistance by directly inhibiting oxidant-activated O00329 and is mimicked by O00329 knockout or by selective inhibitors . Other treatments may also interact in this pathway , making it possible to reverse corticosteroid resistance in patients with P48444 , as well as in smokers with asthma and some patients with severe asthma in whom similar mechanisms operate . Other histone modifications , including methylation , tyrosine nitration , and ubiquitination may also affect histone function and inflammatory gene expression , and better understanding of these epigenetic pathways could led to novel antiinflammatory therapies , particularly in corticosteroid-resistant inflammation . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . Histone deacetylase-2 and airway disease . The increased expression of inflammatory genes in inflammatory lung diseases is regulated by acetylation of core histones , whereas histone deacetylase-2 ( Q92769 ) suppresses inflammatory gene expression . Corticosteroids suppress inflammatory genes in asthma by inhibiting histone acetyltransferase and in particular by recruiting Q92769 to the nuclear factor-kappaB-activated inflammatory gene complex . This involves deacetylation of the acetylated glucocorticoid receptor . In P48444 , severe asthma and asthmatics who smoke , Q92769 is reduced , thus preventing corticosteroids from suppressing inflammation . The reduction in Q92769 appears to be secondary to increased oxidative and nitrative stress in the lungs . Antioxidants and inhibitors of nitric oxide synthesis may therefore restore corticosteroid sensitivity in P48444 , but this can also be achieved by low concentrations of theophylline and curcumin , which act as HDAC activators . DB00277 is a direct inhibitor of oxidant-activated phosphoinositide-3-kinase-delta , which is involved in inactivation of Q92769 . In the future selective O00329 inhibitors and more direct activators of Q92769 may be used to treat corticosteroid-resistant inflammatory diseases of the lung , including P48444 , severe asthma and asthma in smokers . Decreased exercise-induced expression of nuclear factor-κB-regulated genes in muscle of patients with P48444 . BACKGROUND : Nuclear factor ( NF ) -κB activation and oxidative stress are physiologic responses of skeletal muscle to exercise but may be impaired in patients with P48444 . Therefore , we investigated NF-κB activity and expression of NF-κB-regulated genes in muscle of patients with P48444 and control subjects before and after exercise . METHODS : Quadriceps specimens were obtained before , immediately after , and 2 h after a submaximal cycle ergometry test from seven patients with P48444 ( 50.6 ± 5.7 SEM Q99581 (1) of patients with P48444 ) and seven age-matched control subjects . NF-κB DNA-binding activity in muscle and peripheral blood mononuclear cells ( PBMCs ) was determined using electrophoretic mobility shift assay and enzyme-linked immunosorbent assay , respectively . mRNA expression and protein carbonylation were measured by real-time polymerase chain reaction and Western blot , respectively . RESULTS : In control subjects , P05231 , IκBα , tumor necrosis factor-α , IL-1β , superoxide dismutase , thioredoxin , heme oxygenase 1 , and heat shock protein-70 were upregulated in muscle after exercise , whereas in patients with P48444 only P05231 mRNA was increased . Exercise-induced antiapoptotic Bcl2 mRNA levels were attenuated in patients with P48444 compared with control subjects . Basal muscle protein oxidation was higher in patients with P48444 than in control subjects , but attenuated in response to exercise . No exercise-induced changes in NF-κB DNA-binding activity in muscle and PBMCs of either group were detected . CONCLUSIONS : Skeletal muscle of patients with P48444 is characterized by an impaired response to exercise of NF-κB-regulated genes encoding inflammatory cytokines , antioxidants , stress proteins , and survival factors . [ Transport of mucoid mucus in healthy individuals and patients with chronic obstructive pulmonary disease and bronchiectasis ] . OBJECTIVE : To characterise and compare the in vitro transport properties of respiratory mucoid secretion in individuals with no lung disease and in stable patients with chronic obstructive pulmonary disease ( P48444 ) and bronchiectasis . METHODOLOGY : Samples of mucus were collected , from 21 volunteers presenting no lung disease who had undergone surgery , from 10 patients presenting chronic P48444 , and from 16 patients with bronchiectasis . Mucociliary transport ( Q8IVS2 ) , transport by cough ( DB00919 ) , and contact angle ( P62158 ) were evaluated . RESULTS : Q8IVS2 was found to be greater in healthy individuals ( 1.0±0.19 ) than in P48444 ( 0.91±0.17 ) and bronchiectasis ( 0.76±0.23 ) patients ( p < 0.05 ) , whereas DB00919 was greater in P48444 patients ( 16.31±7.35 cm ) than in patients with bronchiectasis ( 12.16±6.64 cm ) and healthy individuals ( 10.50±25.8 cm ) ( p < 0.05 ) . No significant differences were observed between the groups regarding P62158 . CONCLUSION : Mucus from healthy individuals allows better mucociliary transport compared to that from patients with lung diseases . However , the mucus from P48444 patients allows a better transport by coughing , demonstrating that these individuals have adapted to a defence mechanism compared to patients with bronchiectasis , who have impairment in their ciliary and cough transport mechanisms . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . How corticosteroids control inflammation : Quintiles Prize Lecture 2005 . Corticosteroids are the most effective anti-inflammatory therapy for many chronic inflammatory diseases , such as asthma but are relatively ineffective in other diseases such as chronic obstructive pulmonary disease ( P48444 ) . Chronic inflammation is characterised by the increased expression of multiple inflammatory genes that are regulated by proinflammatory transcription factors , such as nuclear factor-kappaB and activator protein-1 , that bind to and activate coactivator molecules , which then acetylate core histones to switch on gene transcription . Corticosteroids suppress the multiple inflammatory genes that are activated in chronic inflammatory diseases , such as asthma , mainly by reversing histone acetylation of activated inflammatory genes through binding of liganded glucocorticoid receptors ( GR ) to coactivators and recruitment of histone deacetylase-2 ( Q92769 ) to the activated transcription complex . At higher concentrations of corticosteroids GR homodimers also interact with DNA recognition sites to active transcription of anti-inflammatory genes and to inhibit transcription of several genes linked to corticosteroid side effects . In patients with P48444 and severe asthma and in asthmatic patients who smoke Q92769 is markedly reduced in activity and expression as a result of oxidative/nitrative stress so that inflammation becomes resistant to the anti-inflammatory actions of corticosteroids . DB00277 , by activating HDAC , may reverse this corticosteroid resistance . This research may lead to the development of novel anti-inflammatory approaches to manage severe inflammatory diseases . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . [ Assessment of the hypothalamic-hypophyseal-adrenal axis in patients with chronic obstructive lung disease. Comparison of inhalant with systemic glucocorticoid therapy ] . OBJECTIVE : The action of inhalation and systemic treatment of chronic obstructive pulmonary disease by suppressing the hypothalamo-hypophyseal-adrenal axis was compared in patients with chronic obstructive pulmonary disease ( P48444 ) . PATIENTS AND METHODS : DB01285 ( DB01285 ) and cortisol concentrations were evaluated after a corticotropin-releasing-hormone ( P06850 ) -test in 50 patients ( aged 43 +/- 14 years ) with chronic obstructive pulmonary disease ( P48444 ) receiving inhalant glucocorticoid treatment ( IGC ) , 61 patients ( aged 54 +/- 11 years ) with P48444 on systemic glucocorticoid treatment ( SGC ) and 50 healthy volunteers ( 32 +/- 4 years ) . RESULTS : All 50 patients on IGC had normal P06850 test results . 30 of 61 patients with SGC had decreased cortisol response ( 12 patients had no and 18 a reduced rise in cortisol ) . DB01285 concentration was lower in patients on IGC than in the control group ( basal DB01285 15.6 pg/ml and 24.5 pg/ml , respectively ; after stimulation 40.3 vs 54.4 pg/ml , respectively ) . But systemic glucocorticoid treatment clearly caused suppression of basal ( 12.1 pg/ml ) and stimulated ( 27.4 pg/ml ) DB01285 levels with correspondingly decreased cortisol levels ( basal : 75.1 and 118.7 ng/ml [ IGC ] , respectively , and after stimulation 128.5 and 225.9 ng/ml ) . CONCLUSIONS : Patients with P48444 on inhalant glucocorticoid treatment have a clearly lower risk of adrenal cortical insufficiency than those on oral glucocorticoid treatment . But some suppression of DB01285 secretion is demonstrable even in the former . Clinical significance of these findings seems unlikely . Development of adrenal cortical insufficiency need not be feared in patients treated with inhalant glucocorticoids . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . P78423 activates a signal transduction pathway similar to Q9H244 and is associated with impaired clopidogrel responsiveness . OBJECTIVE : P78423 ( P78423 ) activates a G(αi) protein-coupled signaling pathway similar to the one activated by ADP via P2Y(12) , which is the drug target of clopidogrel . P78423 levels are increased under several disease conditions associated with impaired clopidogrel responsiveness . METHODS AND RESULTS : Blood samples were obtained from healthy volunteers and from 40 patients under chronic clopidogrel treatment . P78423 reduced prostaglandin E1-induced vasodilator-stimulated phosphoprotein phosphorylation by ≈ 25 % ( P < 0.01 ) at least partially mimicking the effect of ADP via P2Y(12) . In vitro , P78423 increased platelet reactivity index in clopidogrel-treated patients indicating potential activation of downstream targets of P2Y(12) . When stratifying patients by their P78423 levels , patients within the highest quartile of P78423 ( 2042 ± 25 pg/mL ) had the weakest response to clopidogrel ( platelet reactivity index , 68 ± 4 % ) , and patients within the lowest quartile ( 479 ± 50 pg/mL ) had the strongest response ( platelet reactivity index , 48 ± 7 % ; P=0.0106 ) . P78423 by itself induced phosphoinositide 3-kinase activation leading to Akt phosphorylation at DB00133 (473) ( P < 0.01 versus basal ) . CONCLUSIONS : In addition to desensitizing platelets to prostaglandin E1 via G(αi) , P78423 induces phosphoinositide 3-kinase-dependent Akt phosphorylation via a G(βγ) protein similar to ADP signaling through P2Y(12) . P78423 increased the platelet ADP response in clopidogrel-treated patients . Once released from an atherosclerotic lesion , this mechanism could contribute locally to impaired clopidogrel responsiveness at the vulnerable plaque . Simvastatin attenuates experimental small airway remodelling in rats . BACKGROUND AND OBJECTIVE : The aim of this study was to assess the beneficial effects of simvastatin on cigarette smoke-induced small airway remodeling in rats . METHODS : Simvastatin was administered at different doses for 16 weeks to rats with cigarette smoke-induced small airway remodelling . Morphological analyses were performed , and collagen deposition , production of growth factors , inflammatory parameters and RhoA , as well as the Smad signalling pathway in the lungs , were examined . RESULTS : Simvastatin attenuated small airway wall thickening and prevented the increase in lung hydroxyproline content and collagen deposition induced in airway walls by cigarette smoking . In addition , simvastatin downregulated transforming growth factor-beta1 and connective tissue growth factor protein and gene expression in the lungs . Furthermore , accumulation of macrophages and neutrophils and increases in tumour necrosis factor-alpha concentration in BAL fluid were inhibited by simvastatin . Simultaneously , the expression of RhoA and the phosphorylation of Q15796 and P84022 in lungs exposed to cigarette smoke were inhibited during simvastatin administration . However , the increased expression of Q15796 and P84022 proteins and the decreased level of O15105 protein in remodelled lungs were not affected by simvastatin . CONCLUSIONS : Simvastatin attenuated experimental small airway remodelling , as indicated by decreases in collagen deposition and small airway wall thickening . Simvastatin may inhibit cigarette smoke-induced small airway remodelling by reducing growth factor expression and inflammation . The mechanism of action of simvastatin on small airway remodelling involved RhoA and the Smad signalling pathway . These findings indicate that simvastatin may have potential beneficial effects in the treatment of P48444 . Role of phosphoinositide 3-kinase beta in platelet aggregation and thromboxane A2 generation mediated by Gi signalling pathways . PI3Ks ( phosphoinositide 3-kinases ) play a critical role in platelet functional responses . PI3Ks are activated upon Q9H244 receptor stimulation and generate pro-aggregatory signals . Q9H244 receptor has been shown to play a key role in the platelet aggregation and thromboxane A2 generation caused by co-stimulation with Gq or Gz , or super-stimulation of Gi pathways . In the present study , we evaluated the role of specific PI3K isoforms alpha , beta , gamma and delta in platelet aggregation , thromboxane A2 generation and P29323 ( extracellular-signal-regulated kinase ) activation . Our results show that loss of the PI3K signal impaired the ability of ADP to induce platelet aggregation , P29323 phosphorylation and thromboxane A2 generation . We also show that Gq plus Gi- or Gi plus Gz-mediated platelet aggregation , P29323 phosphorylation and thromboxane A2 generation in human platelets was inhibited by O43548 -221 , a P42338 -selective inhibitor , but not by PIK75 ( a P42336 inhibitor ) , AS252424 ( a P48736 inhibitor ) or IC87114 ( a O00329 inhibitor ) . O43548 -221 also showed a similar inhibitory effect on the Gi plus Gz-mediated platelet responses in platelets from P47900 -/- mice . Finally , 2MeSADP ( 2-methyl-thio-ADP ) -induced Akt phosphorylation was significantly inhibited in the presence of O43548 -221 , suggesting a critical role for P42338 in Gi-mediated signalling . Taken together , our results demonstrate that P42338 plays an important role in ADP-induced platelet aggregation . Moreover , P42338 mediates ADP-induced thromboxane A2 generation by regulating P29323 phosphorylation . Role of the P78423 - P49238 axis in chronic inflammatory lung diseases . Persistent inflammation is often present in patients with lung diseases such as chronic obstructive pulmonary diseases ( P48444 ) and pulmonary hypertension . Circulatory leukocyte migration through the lung vascular endothelium contributes to the structural destruction and remodeling seen in these chronic lung diseases . An inflammatory chemokine P78423 /fractalkine is associated with inflammatory lung diseases . Membrane-anchored P78423 serves as an adhesion molecule to capture subsets of mononuclear leukocytes that express the sole receptor , P49238 . The extracellular chemokine domain of P78423 can be cleaved/shed by a disintegrin and metalloproteinase domain ( ADAM ) from stimulus-exposed cells . Soluble P78423 chemoattracts and activates P49238 + leukocytes such as CD8+ , P01730 + , and γδ T lymphocytes , natural killer cells , dendritic cells , and monocytes/macrophages . P49238 + leukocyte attachment to and migration through the lung vascular endothelium lead to mononuclear cell accumulation in the lung vessel walls and parenchyma . Infiltrated P49238 + immune cells can release mediators to induce injury , stimulate proliferation , and/or chemoattract inflammatory cells . This contributes to structural destruction and remodeling in the development of inflammatory lung diseases . Limited clinical success in treating chronic pulmonary diseases-associated lung functional decline indicates the urgency and significance of understanding upstream signaling that triggers inflammation . This article reviews the advances in the P78423 - P49238 axis-mediated modulation of mononuclear leukocyte adhesion and migration in inflammatory lung diseases such as P48444 and pulmonary hypertension . Better understanding of the constant flow of circulating leukocytes into the lung vessel wall and parenchyma will help set a stage for the development of novel therapeutic approaches to treat or even cure chronic lung diseases including P48444 and pulmonary hypertension .

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The clinical effectiveness of DB00315 in the acute treatment of migraine . Efficacy with currently marketed antimigraine compounds is less than optimal . DB00315 is a novel and selective P28221 receptor agonist in development for the acute treatment of migraine . It shows evidence of both central and peripheral activity within the trigemino-vascular system and it is rapidly absorbed following oral administration . In clinical studies in migraine patients , a headache response at 2 hours has been observed in 65-81 % of patients at doses above 1 mg . Favourable response rates are reported as early as 1 hour post-dose and efficacy rates continue to improve up to 4 hours . Headache recurrence is reported by 25-35 % of patients and DB00315 is also effective in relieving the non-headache symptoms of migraine . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . DB00315 , a new central and peripherally acting P28221 receptor agonist in the acute oral treatment of migraine : a double-blind , placebo-controlled , dose-range finding study . DB00315 is a novel , centrally and peripherally , acting 5-hydroxytryptamine1D receptor agonist . We investigated the efficacy and safety of 1 , 5 , and 25 mg of oral DB00315 in the acute treatment of migraine in a randomized , double-blind , placebo-controlled , parallel-group clinical trial involving 84 patients . The proportion of patients in whom the headache improved within 2 hours from moderate or severe to mild or no pain ( primary efficacy measure ) was 15 % for placebo-treated patients and 27 % ( 1 mg ) , 62 % ( 5 mg ) , and 81 % ( 25 mg ) for patients treated with DB00315 . Treatment differences compared with placebo were 12 % ( 95 % CI - 12 , 37 ; p = 0.460 ) for 1 mg DB00315 , 47 % ( CI 21 , 73 ; p < 0.005 ) for 5 mg DB00315 , and 66 % ( CI 43,89 ; p < 0.001 ) for 25 mg DB00315 . Photophobia and nausea also showed improvement after DB00315 . Adverse events were generally mild and transient in all treatment groups . There were no clinically significant changes in ECG recordings , blood pressure , or laboratory tests . Oral DB00315 ( 5 and 25 mg ) is highly effective and well tolerated in the acute treatment of migraine . The response rates and treatment differences compared with placebo in this study suggest possible superiority over existing antimigraine therapies . This needs to be confirmed in formal comparative trials . Inhibition of the trigemino-vascular system with P28221 agonist drugs : selectively targeting additional sites of action . Inappropriate activation of the trigemino-vascular system is thought to be important in the pathogenesis of a migraine attack . The P28221 agonist sumatriptan , which is highly effective in the acute treatment of migraine , inhibits trigemino-vascular activation in animals , although its actions are normally limited to peripheral components of the trigemino-vascular system . DB00315 , a novel P28221 agonist drug , which is also highly effective in the acute treatment of migraine , acts not only at these sites , but , additionally within the brainstem , inhibiting trigemino-vascular activation centrally as well as peripherally . This article describes the pre-clinical development of DB00315 and considers , specifically , the approaches taken in the design of a molecule with attributes which facilitate access to brainstem components of the trigeminal pathway and combine this with good oral bioavailability . Clinical safety of DB00315 : aggregated data from patients and volunteers to date . The tolerability of DB00315 , a novel , selective and highly effective P28221 receptor agonist in development for the acute treatment of migraine , has been evaluated in a number of clinical pharmacology and patient studies across the dose range 1-50 mg . DB00315 has been well tolerated across the entire dose range and no clinically relevant changes in routine laboratory parameters , blood pressure or ECG recordings have been observed . Adverse experiences reported are generally dose related , mild to moderate and resolve spontaneously . Chest-related symptoms occur infrequently and the cardiovascular safety profile of DB00315 is considered particularly favourable . DB00315 , therefore , possesses a desirable safety profile which is well suited to broad-based outpatient administration . GLC756 decreases P01375 via an alpha2 and beta2 adrenoceptor related mechanism . GLC756 , a polyvalent anti-glaucoma drug showed in an endotoxin-induced-uveitis model ( EIU ) in rats a significant tumor necrosis factor-alpha ( P01375 ) decrease in serum , indicating an additional anti-inflammatory potential of this compound . The receptors on which GLC756 binds ( D1 , D2 , D4 , alpha-1 , alpha-2 , P08908 , P28335 , P28221 , 5-HT2 A , beta-1 , and beta-2 ) were suggested to play a role . In order to identify a receptor type mediating the P01375 lowering response , GLC756 was combined with various counteracting compounds ( CP ) . For EIU , 8-week-old Lewis rats were intravenously injected at 160 microg lipopolysaccharide ( LPS ) from Salmonella typhimurium . Before EIU-induction animals received either one of the CP 's or GLC756 alone , or GLC756 in combination with one of the CP 's . P01375 was determined in serum 2h post EIU-induction . Treatment with CP 's alone indicated that agonistic effects on beta-2 adrenoceptors and antagonistic effects on alpha-2 , P08908 and P28221 receptors resulted in statistically significant decreased P01375 levels in comparison to the LPS-control group . In combination with GLC756 , the counteracting CP 's domitor ( alpha-2 adrenoceptor agonist ) and ICI 118551 ( beta-2 adrenoceptor antagonist ) inhibited completely the P01375 decreasing effect of GLC756 . Counteracting the P08908 receptor with the P08908 agonist 8-OH-DPAT could not prevent the P01375 decreasing effect of GLC756 . In conclusion , the antagonistic effect on alpha-2 adrenoceptors and the agonistic effect on beta-2 adrenoceptors were identified as mechanism for the P01375 decreasing effect of GLC756 . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . The tolerability and pharmacokinetics of the novel antimigraine compound DB00315 in healthy male volunteers . 1. DB00315 is a novel and selective agonist at P28221 receptors , with central and peripheral actions , currently in development for the acute oral treatment of migraine . 2 . The pharmacokinetic and tolerability profiles of single oral doses from 1-50 mg DB00315 were investigated in 12 healthy male volunteers in a double-blind , placebo-controlled , dose-escalating study . 3 . DB00315 was well tolerated with most adverse experiences of mild and transient nature . 4 . Absorption was rapid with dose-independent kinetics . Median tmax was 2-4 h although 50-85 % of eventual Cmax was attained within 1 h . The t1/2 was 2.5-3 h with a high apparent plasma clearance ( CL/F > 2000 ml min-1 ) and apparent volume of distribution ( Vz/F ) of 400-500 l . 5 . Three metabolites were detected in plasma and urine , one of which , the N-desmethyl metabolite , has P28221 agonist activity . 6 . DB00315 showed no clinically significant effects on blood pressure , heart rate , ECG or laboratory variables at any dose and demonstrated a tolerability and pharmacokinetic profile compatible with an acute oral migraine treatment . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . Activation of the JAK/ P35610 pathway in vascular smooth muscle by serotonin . Serotonin ( 5-hydroxytryptamine , 5-HT ) is a vasoconstrictor and mitogen whose levels are elevated in diabetes . Previous studies have shown the presence of 5- Q13049 , P41595 , and P28222 receptors in vascular smooth muscle cells ( VSMCs ) . There are currently no data regarding P41595 and P28222 receptor activation of the JAK/ P35610 pathway in VSMCs and resultant potential alterations in 5-HT signaling in diabetes . Therefore , we tested the hypothesis that 5-HT differentially activates the JAK/ P35610 pathway in VSMCs under conditions of normal ( 5 mM ) and high ( 25 mM ) glucose . Treatment of rat VSMCs with 5-HT ( 10(-6) M ) resulted in time-dependent activation ( approximately 2-fold ) of O60674 , P23458 , and P42224 , but not P40763 ( maximal at 5 min , returned to baseline by 30 min ) . The P41595 receptor agonist BW723C86 and the P28222 receptor agonist CGS12066A ( 10(-9)-10(-5) M , 5-min stimulation ) did not activate the JAK/ P35610 pathway . Treatment with the 5- Q13049 receptor antagonist ketanserin ( 10 nM ) inhibited O60674 activation by 5-HT . Treatment of streptozotocin-induced diabetic rats with ketanserin ( 5 mg.kg-1.day-1 ) reduced activation of O60674 and P42224 but not P40763 in endothelium-denuded thoracic aorta in vivo . 5-HT ( 10(-6) M ) treatment resulted in increased cell proliferation and increased DNA synthesis , which were inhibited by the O60674 inhibitor AG490 . Further studies with apocynin , diphenyleneiodonium chloride , catalase , and virally transfected superoxide dismutase had no effect at either glucose concentration on activation of the JAK/ P35610 pathway by 5-HT . Therefore , we conclude that 5-HT activates O60674 , P23458 , and P42224 via the 5- Q13049 receptors in a reactive oxygen species-independent manner under both normal and high glucose conditions . DB00175 -induced proangiogenic effects depend upon extracellular P09038 . The P04035 inhibitors ( statins ) have been shown to exert several protective effects on the vasculature that are unrelated to changes in the cholesterol profile , and to induce angiogenesis . The proangiogenic effect exerted by statins has been attributed to the activation of the PI3K/Akt pathway in endothelial cells ; however , it is unclear how statins activate this pathway . DB00175 -mediated activation of Akt and MAPK occurs rapidly ( within 10 min. ) and at low doses ( 10 nM ) . Here , we hypothesized that P09038 contributes to the proangiogenic effect of statins . We found that pravastatin , a hydrophilic statin , induced phosphorylation of the FGF receptor ( FGFR ) in human umbilical vein endothelial cells . SU5402 , an inhibitor of FGFR , abolished pravastatin-induced PI3K/Akt and MAPK activity . Likewise , anti- P09038 function-blocking antibodies inhibited Akt and MAPK activity . Moreover , depletion of extracellular P09038 by heparin prevented pravastatin-induced phosphorylation of Akt and MAPK . Treatment with P09038 antibody inhibited pravastatin-enhanced endothelial cell proliferation , migration and tube formation . These observations indicate that pravastatin exerts proangiogenic effects in endothelial cells depending upon the extracellular P09038 . Serotonin via P34969 receptors activates p38 mitogen-activated protein kinase and protein kinase C epsilon resulting in interleukin-6 synthesis in human U373 MG astrocytoma cells . Serotonin [ 5-hydroxytryptamine ( 5-HT ) ] is a widely distributed neurotransmitter which is involved in neuroimmunomodulatory processes . Previously , it has been demonstrated that 5-HT may induce interleukin ( IL ) -6 expression in primary rat hippocampal astrocytes . The present study was undertaken to investigate the molecular pathways underlying this induction of P05231 synthesis . As a model system , we used the human astrocytoma cell line U373 MG , which synthesizes P05231 upon stimulation with various inducers . 5-HT dose- and time-dependently induced P05231 protein synthesis . We identified several 5-HT receptors to be expressed on U373 MG cells , including the P28221 , 5- Q13049 , 5- Q9H205 and P34969 receptors . In this report , we show that the 5-HT-induced P05231 release is mediated by the P34969 receptor based on several agonist/antagonists that were used . 5-HT-induced P05231 synthesis is inhibited by the partially selective P34969 receptor antagonist , pimozide , and the selective antagonist SB269970 . Furthermore , P05231 synthesis was induced by the P34969 receptor agonist carboxamidotryptamin . In addition , we found p38 MAPKs and protein kinase C ( PKC ) epsilon to be involved in 5-HT-induced P05231 synthesis as specific inhibitors of these enzymes ( SB202190 and RO-31-8425 , respectively ) blocked 5-HT-induced P05231 synthesis . Furthermore , 5-HT mediated the phosphorylation of both p38 MAPK as well as the PKC epsilon isoform . The Q8NFH3 /44 MAPKs , however , were not involved in 5-HT-induced P05231 synthesis . This study shows , for the first time , a central role of P34969 receptor linked to p38 MAPK and PKC epsilon for the induction of cytokine synthesis in astrocytic cells . Acute migraine therapy : the newer drugs . In 1996 , our knowledge of acute antimigraine therapy expanded in three major areas . First , large surveys have confirmed the remarkable efficacy profile of sumatriptan in clinical practice . No satisfying clinical , pharmacokinetic or genetic explanations were found for its major shortcomings : nonresponders , headache recurrence and noncardiac chest symptoms . Second , the novel P28222 /D agonists zolmitriptan ( DB00315 ) , rizatriptan ( MK-462 ) , eletriptan ( UK-116,044 ) , avitriptan ( BMS-180048 ) and alniditan ( R091274 ) were all proved superior to placebo for attack treatment , but their advantages over sumatriptan are yet to be analysed in more detail . A higher lipophilicity explains ( except for alniditan ) their greater oral bioavailability and better central nervous system penetration . A central action now proved experimentally in animals and in humans for P28222 /D agonists such as zolmitriptan may be advantageous for the antimigraine efficacy , but it could also increase sedation . Third , an endothelin ( Ro470203 , DB00559 ) and a neurokinin 1 ( RPR100893 ) receptor antagonist were found to be ineffective in migraine . Both compounds are potent inhibitors of neurogenic plasma extravasation in rat dura mater , which might suggest that this pharmacological property does not necessarily predict efficacy in aborting migraine attacks . Structure functional expression and spatial distribution of a cloned cDNA encoding a rat P28221 -like receptor . Using polymerase chain reaction ( PCR ) a complementary DNA ( cDNA ) encoding a 5-hydroxytryptamine ( 5-HT ) receptor was isolated from rat forebrain . The amplified cDNA specifies an open reading frame of 374 amino acids comprising seven putative transmembrane regions . Expression of the cloned cDNA in human embryonic kidney cells ( P29320 293 ) was used to establish the pharmacological profile of the encoded receptor polypeptide . Membranes containing the cloned receptor showed high affinity binding of [ 3H ] -5-HT . Competition binding experiments with a variety of serotonin receptor ligands displayed a rank order of affinities corresponding to a P28221 subtype : 5-CT > 5-HT = metergoline > CGS 12066 > methysergide > sumatriptan > mianserin = (-)alpha-Me-5-HT = yohimbine > 8-OH-DPAT > or = rauwolscine > spiperone > DOI > propranolol > or = 2-Me-5-HT > or = ICS 205930 . Ketanserin and ritanserin displaced [ 3H ] -5-HT-binding in a biphasic manner . In situ hybridization revealed highest expression of the corresponding mRNA in the pyramidal layer of the olfactory tubercle and the nucleus caudatus and accumbens . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Mobilization of Ph chromosome-negative peripheral blood stem cells in chronic myeloid leukaemia patients with imatinib mesylate-induced complete cytogenetic remission . Imatinib mesylate ( IM , DB00619 , Glivec ) can induce a high rate of complete cytogenetic response ( CCR ) in chronic myeloid leukaemia ( CML ) patients , although to date the majority of patients continue to have detectable disease by sensitive reverse transcription polymerase chain reaction ( RT-PCR ) . It is therefore possible that these patients may ultimately relapse and require treatment such as autologous peripheral blood stem cell transplant ( APBSCT ) . We attempted mobilization of haemopoietic progenitor cells from 58 patients in CCR using recombinant human granulocyte colony-stimulating factor [ rHu- DB00099 ; 10 micro g/kg/d subcutaneously ( s.c. ) for at least 4 d ] alone , while continuing IM treatment . The median d 5 ( peak ) P28906 + count was 11.5/ microl ( range 0-108/ microl ) , and 43/58 ( 74 % ) patients underwent a median of two ( range 1-3 ) apheresis procedures . A median dose of 2.1 x 10(6)/kg P28906 + cells ( range 0.1-6.5 x 10(6)/kg ) was collected . Some 84 % of 31 collections analysed were negative for the Philadelphia ( Ph ) chromosome or breakpoint cluster region and Abelson murine leukaemia viral oncogene homologue ( P11274 - P00519 ) translocation by cytogenetics or fluorescent in situ hybridization respectively . No toxicity was reported with the regimen . Overall , the target P28906 + dose ( 2 x 10(6)/kg P28906 + ) was attained in 23/58 ( 40 % ) patients who entered the study . In summary , we have demonstrated that successful mobilization of Ph- P28906 + cells from IM-treated patients in CCR is possible using rHu-G- P04141 alone . Inhibition of c-kit receptor tyrosine kinase activity by DB00619 , a selective tyrosine kinase inhibitor . DB00619 ( formerly known as CGP 57148B ) is a known inhibitor of the c-abl , bcr-abl , and platelet-derived growth-factor receptor ( P09619 ) tyrosine kinases . This compound is being evaluated in clinical trials for the treatment of chronic myelogenous leukemia . We sought to extend the activity profile of DB00619 by testing its ability to inhibit the tyrosine kinase activity of c-kit , a receptor structurally similar to P09619 . We treated a c-kit expressing a human myeloid leukemia cell line , M-07e , with DB00619 before stimulation with Steel factor ( SLF ) . DB00619 inhibited c-kit autophosphorylation , activation of mitogen-activated protein ( Q96HU1 ) kinase , and activation of Akt without altering total protein levels of c-kit , Q96HU1 kinase , or Akt . The concentration that produced 50 % inhibition for these effects was approximately 100 nmol/L . DB00619 also significantly decreased SLF-dependent growth of M-07e cells in a dose-dependent manner and blocked the antiapoptotic activity of SLF . In contrast , the compound had no effect on Q96HU1 kinase activation or cellular proliferation in response to granulocyte-macrophage colony-stimulating factor . We also tested the activity of DB00619 in a human mast cell leukemia cell line ( HMC-1 ) , which has an activated mutant form of c-kit . DB00619 had a more potent inhibitory effect on the kinase activity of this mutant receptor than it did on ligand-dependent activation of the wild-type receptor . These findings show that DB00619 selectively inhibits c-kit tyrosine kinase activity and downstream activation of target proteins involved in cellular proliferation and survival . This compound may be useful in treating cancers associated with increased c-kit kinase activity . 5-HT2 receptor involvement in conditioned olfactory learning in the neonate rat pup . These experiments addressed the role of 5-HT2 receptors in conditioned olfactory learning . Ritanserin , a 5- Q13049 /2C antagonist , was injected subcutaneously into postnatal day ( P01160 ) 7 pups before or after conditioned olfactory training to a peppermint odor . When the pups were tested for olfactory preference on P01160 8 , those injected with ritanserin before training failed to acquire an odor preference whereas those injected after training learned . This suggested that the 5-HT2 receptor is required only in the acquisition of conditioned olfactory learning . Injection of ritanserin directly into the olfactory bulb before training also blocked preference for the peppermint odor . In pups that had depletion of the 5-HT input to the bulb , subcutaneous injection of a 5- Q13049 /2C agonist was sufficient to maintain conditioned olfactory learning , confirming the importance of 5-HT in learning . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . Mapping of the serotonin P28221 beta autoreceptor gene on chromosome 6 and direct analysis for sequence variants . Abnormal brain serotonin function may be characteristic of several neuropsychiatric disorders . Thus , it is important to identify polymorphic genes and screen for functional variants at loci coding for genes that control normal serotonin functions . P28221 beta is a terminal serotonin autoreceptor which may play a role in regulating serotonin synthesis and release . Using an SSCP technique we screened for P28221 beta coding sequence variants in psychiatrically interviewed populations , which included controls , alcoholics , and alcoholic arsonists and alcoholic violent offenders with low P04141 concentrations of the main serotonin metabolite 5-HIAA . A common polymorphism was identified in the P28221 beta gene with allele frequencies of 0.72 and 0.28 . The SSCP variant was caused by a silent G to C substitution at nucleotide 861 of the coding region . This polymorphism could also be detected as a HincII RFLP of amplified DNA . DNAs from informative CEPH families were typed for the HincII RFLP and analyzed with respect to 20 linked markers on chromosome 6 . Multipoint analysis placed the P28221 beta receptor gene between markers D6S286 and D6S275 . A maximum two-point lod score of 10.90 was obtained to D6S26 , which had been previously localized on 6q14-15 . Chromosomal aberrations involving this region have been previously shown to cause retinal anomalies , developmental delay , and abnormal brain development . This region also contains the gene for North Carolina-type macular dystrophy . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Different cholinesterase inhibitor effects on P04141 cholinesterases in Alzheimer patients . BACKGROUND : The current study aimed to compare the effects of different cholinesterase inhibitors on acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) activities and protein levels , in the cerebrospinal fluid ( P04141 ) of Alzheimer disease ( AD ) patients . METHODS AND FINDINGS : AD patients aged 50-85 years were randomized to open-label treatment with oral rivastigmine , donepezil or galantamine for 13 weeks . P22303 and BuChE activities were assayed by Ellman 's colorimetric method . Protein levels were assessed by enzyme-linked immunosorbent assay ( ELISA ) . Primary analyses were based on the Completer population ( randomized patients who completed Week 13 assessments ) . 63 patients were randomized to treatment . DB00989 was associated with decreased P22303 activity by 42.6 % and decreased P22303 protein levels by 9.3 % , and decreased BuChE activity by 45.6 % and decreased BuChE protein levels by 21.8 % . DB00674 decreased P22303 activity by 2.1 % and BuChE activity by 0.5 % , but increased P22303 protein levels by 51.2 % and BuChE protein levels by 10.5 % . Donepezil increased P22303 and BuChE activities by 11.8 % and 2.8 % , respectively . Donepezil caused a 215.2 % increase in P22303 and 0.4 % increase in BuChE protein levels . Changes in mean P22303 -Readthrough/Synaptic ratios , which might reflect underlying neurodegenerative processes , were 1.4 , 0.6 , and 0.4 for rivastigmine , donepezil and galantamine , respectively . CONCLUSION : The findings suggest pharmacologically-induced differences between rivastigmine , donepezil and galantamine . DB00989 provides sustained inhibition of P22303 and BuChE , while donepezil and galantamine do not inhibit BuChE and are associated with increases in P04141 P22303 protein levels . The clinical implications require evaluation . Differential functional activity of 5-hydroxytryptamine receptor ligands and beta adrenergic receptor antagonists at 5-hydroxytryptamine1B receptor sites in Chinese hamster lung fibroblasts and opossum renal epithelial cells . Functional activity of 5-hydroxytryptamine ( 5-HT ) receptor ligands and beta adrenergic receptor antagonists was studied at P28222 receptor sites in Chinese hamster lung ( CHL ) fibroblasts by measuring two cellular responses : inhibition of forskolin-stimulated cyclic AMP formation and potentiation of basic fibroblast growth ( BFGF ) induced mitogenesis . A good correlation was found between the potency of agonists to inhibit forskolin-induced cyclic AMP formation and their potency to potentiate P09038 -induced thymidine incorporation in CHL fibroblasts . Potent agonist activity was measured with 5-methoxy-3,1,2,3,6-tetrahydro-4-pyidinyl- 1H-indole ( RU 24,969 ) , 5-carboxamidotryptamine ( 5-CT ) , 3-(1,2,5,6)-tetrahydro-4-pyridyl-5-pyrrolo(3,2-b)pyril-5-one ( CP 93,129 ) and 5-HT , whereas sumatriptan displayed weak agonist activity at concentrations different from its binding affinity for P28222 binding sites . In contrast to the observed P28222 receptor-mediated agonist activity in opossum kidney cells for metergoline and the beta adrenergic receptor antagonists : cyanopindolol , 4-(3-tert-butyl-amino-2-hydroxypropoxy)-indole-2 carbonic acid isopropyl ester ( SDZ 21,009 ) , isamoltane , (-)-propranolol and (-)-pindolol , antagonist activity at P28222 receptor sites was yielded in CHL fibroblasts in accordance with the reported observations at rat brain P28222 receptors . Methiothepin was the only compound that antagonized both the opossum kidney cell and CHL fibroblast P28222 receptor-mediated responses although the antagonist effect was more pronounced in CHL fibroblasts. ( ABSTRACT TRUNCATED AT 250 WORDS ) Evaluation of the long-term safety and efficacy of DB00315 in the treatment of migraine . DB00315 is an orally active P28221 agonist with both central and peripheral actions that is currently being developed as an acute antimigraine treatment . Several studies have demonstrated the safety and efficacy of DB00315 in the treatment of a single migraine headache . The objectives of this open study are to assess the safety and efficacy of DB00315 when used for a period of up to one year . Patients can treat as many migraine headaches as desired with an oral treatment regimen of DB00315 . An initial 5 mg dose for treatment of the migraine headache may be followed with a second 5 mg dose to treat recurrence should it develop . Safety assessments include electrocardiograms , the frequency , intensity and duration of adverse experiences , and routine haematology , urinalysis and clinical chemistry measures . Data presented here are an interim view of the database as of August 1995 and should be considered as preliminary observations . No clinically significant serious adverse experiences have been reported . The adverse experience and efficacy profile appears to be consistent with previous DB00315 studies and this dosing regimen of DB00315 was well tolerated during multiple exposures . Notably , response rates are as good after both initial and repeated exposure ( up to 5 migraines ) . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Effects of PNU-109,291 , a selective P28221 receptor agonist , on electrically induced dural plasma extravasation and capsaicin-evoked c-fos immunoreactivity within trigeminal nucleus caudalis . We studied the effects of PNU-109291 [ ( S ) -(-)-1-[2-[4-(4-methoxyphenyl)-1-piperazinyl]ethyl]-N-methyl-isoc hroman-6-carboxamide ] , a receptor agonist showing 5000-fold selectivity for primate P28221 versus P28222 receptors ( Ennis et al. , J. Med. Chem. 41 , 2180-2183 ) , on dural neurogenic inflammation and on c-fos like immunoreactivity within trigeminal nucleus caudalis evoked by electrical and chemical activation of trigeminal afferents , respectively . Subcutaneous injection of PNU-109291 in male guinea pigs dose-dependently reduced dural extravasation of [ 125I ] -labeled bovine serum albumin evoked by trigeminal ganglion stimulation with an IC50 of 4.2 nmol kg(-1) . A dose of 73.3 nmol kg(-1) blocked the response completely . The selective P28222 /1D receptor antagonist GR-127935 ( > or = 2 micromol kg(-1) i.v. ) prevented this effect . In addition , the number of c-fos immunoreactive cells within guinea pig trigeminal nucleus caudalis induced by chemical meningeal stimulation ( intracisternally administered capsaicin ) was reduced by more than 50 % with PNU-109291 ( > or = 122.2 nmol kg(-1) administered s.c. 45 min before and 15 min after capsaicin ) . These data indicate that the P28221 receptor subtype plays a significant role in suppressing meningeal neurogenic inflammation and attenuating trigeminal nociception in these guinea pig models . Since P28221 receptor mRNA and protein are expressed in trigeminal ganglia but not vascular smooth muscle , the P28221 receptor subtype may become a useful therapeutic target for migraine and related headaches . Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor . Mouse pluripotent hematopoietic stem cells ( PHSC ) were fractionated based on size and density using counterflow centrifugal elutriation ( CCE ) . These heterogeneous PHSC populations were further enriched by subtraction of cells with lineage-specific markers ( Lin- ) followed by positive sorting for c-kit expression . The cells were characterized for their functional and biochemical properties . We defined a subpopulation of c-kit-positive cells that expressed high numbers of c-kit receptors ( c-kitBR ) . One hundred c-kitBR cells from either low- or higher-density fractions were sufficient to repopulate the lymphohematopoietic system in WBB6F1-W/Wv ( W/Wv ) recipients , whereas no PHSC were found in cells with low ( c-kitDULL ) or no ( c-kitNEG ) c-kit expression . Lin- c-kitBR cells were separated into RhoDULL and RhoBR subsets based on their ability to efflux rhodamine 123 ( Rho ) . The PHSC were concentrated in Lin- c-kitBR RhoDULL cells and the number of Lin- c-kitBR RhoBR cells correlated directly with the number of day 12 colony-forming unit-spleen ( CFU- P28222 ) in each fraction . We were not able to enrich further for PHSC using monoclonal antibodies to the cell-surface markers AA4.1 or P01730 , which have been used by others to isolate PHSC . The small , low-density Lin- c-kitBR subset contained PHSC and few CFU- P28222 . This enabled us to assay PHSC for expression of the flk-2 gene , which encodes a tyrosine kinase receptor present on fetal liver PHSC . Purified RNA from the low-density Lin- c-kitBR subset did not contain flk-2 mRNA . We suggest that AA4.1 , P01730 and flk-2 are expressed as stage-specific markers on PHSC in cell cycle . Concentrations of pravastatin and lovastatin in cerebrospinal fluid in healthy subjects . The capability of pravastatin and lovastatin , P04035 inhibitors likely to be taken chronically for hypercholesterolemia , to cross the blood-brain barrier was investigated in normal male volunteers . DB00227 , which is lipophilic , was detected in cerebrospinal fluid ( P04141 ) at concentrations that may have a pharmacologic effect . DB00175 , which is hydrophilic , was not detected in P04141 . It is concluded that pravastatin may have less potential for causing CNS-related side effects than lovastatin . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB00989 improves hippocampal neurogenesis and depression-like behaviors via P08908 receptor stimulation in olfactory bulbectomized mice . DB00989 is a non-competitive inhibitor of both acetylcholinesterase ( P22303 ) and butylcholinesterase ( BuChE ) used to treat mild to moderate dementia in Alzheimer 's disease ( AD ) patients . Although rivastigmine reportedly ameliorates cognitive dysfunction in these patients , its ability to improve Behavioral and Psychological Symptoms of Dementia ( BPSD ) remains unclear . To determine whether rivastigmine treatment antagonizes depression-like behaviors , we chronically administered rivastigmine ( 0.1-1.0mg/kg ) to olfactory bulbectomized ( OBX ) mice once a day for 2weeks , starting 2weeks after bulbectomy . Chronic treatment at 0.3 or 1.0mg/kg dose dependently and significantly improved depression-like behaviors , as assessed by tail suspension ( Q16762 ) , forced swim ( P19883 ) , locomotion and novelty-suppressed feeding ( NSFT ) tests . Importantly , co-administration with WAY-100635 ( 1.0mg/kg ) , a P08908 receptor antagonist , but not ketanserin ( 1.0mg/kg , ) , a 5- Q13049 receptor antagonist , completely blocked rivastigmine-induced anti-depressive effects , suggesting that P08908 receptor stimulation mediates this activity . Consistent with this observation , rivastigmine treatment significantly rescued impaired neurogenesis observed in OBX mice in a P08908 receptor-dependent manner . Furthermore , enhanced protein kinase B ( Akt ) and extracellular signal-regulated kinase ( P29323 ) phosphorylation seen following rivastigmine treatment was closely associated with improved neurogenesis . These effects were blocked by WAY-100635 but not ketanserin treatment . Finally , we confirmed that P08908 but not 5- Q13049 receptor stimulation by specific agonists mimicked rivastigmine-induced anti-depression activity and promoted hippocampal neurogenesis . We conclude that , in addition to enhancing the cholinergic system , rivastigmine treatment restores normal function of the hippocampal serotonergic system , an activity that likely ameliorates depressive behaviors in AD patients . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Computer-aided design and synthesis of 5-substituted tryptamines and their pharmacology at the P28221 receptor : discovery of compounds with potential anti-migraine properties . The design and synthesis of a series of novel 5-substituted tryptamines with pharmacological activity at P28221 and other monoamine receptors is described . Structural modifications of N- and C-linked ( principally hydantoin ) analogues at the 5-position were synthesized and their pharmacological activities were utilized to deduce significant steric and electrostatic requirements of the P28221 and 5- Q13049 receptor subtypes . Conformations of the active molecules were computed which , when overlaid , suggested a pharmacophore hypothesis which was consistent with the affinity and selectivity measured at P28221 and 5- Q13049 receptors . This pharmacophore is composed of a protonated amine site , an aromatic site , a hydrophobic pocket , and two hydrogen-bonding sites . A " selectivity site " was also identified which , if occupied , induced sensitivity for P28221 over 5- Q13049 in this series of molecules . The development and use of the pharmacophore models in compound design is described . In addition , the physicochemical constraints of molecular size and hydrophobicity required for efficient oral absorption are discussed . Utilizing the pharmacophore model in conjunction with the physicochemical constraints of molecular size and log DpH7.4 led to the discovery of DB00315 ( 6 ) , a new selective P28221 agonist with good oral absorption and potential use in the treatment of migraine . P40189 -linked signal transduction promotes the differentiation and maturation of dendritic cells . In order to explore the role of P40189 -linked signal transduction in the differentiation and maturation of dendritic cells ( DC ) , the mAb , B- P28222 , an agonist of P40189 , was used for the activation of P40189 on DC . The effects of cytokines and of anti- P40189 mAb on the proliferation of DC , and their expression of IL-12 and P33681 ( P33681 -1 ) by DC were evaluated . DC differentiating from peripheral blood mononuclear cells did not express the P05231 receptor alpha chain , but expressed P40189 . Anti- P40189 mAb promoted the proliferation of DC , induced by P05112 and granulocyte macrophage colony stimulating factor ( GM- P04141 ) , by up-regulating the GM- P04141 receptor on DC . DC induced by P40189 mAb and cytokines expressed DC-derived CC chemokine , as measured by RT-PCR . Induced DC also stimulated strong proliferation of autologous T cells in mixed lymphocyte reaction since an up-regulated expression of IL-12 and P33681 ( P33681 -1 ) was observed in DC activated by anti- P40189 mAb . Thus , P40189 signal transduction is important for the differentiation and maturation of DC . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Interaction of calmodulin with the serotonin 5-hydroxytryptamine2A receptor . A putative regulator of G protein coupling and receptor phosphorylation by protein kinase C . The 5-hydroxytryptamine2A ( 5- Q13049 ) receptor is a G(q/11)-coupled serotonin receptor that activates phospholipase C and increases diacylglycerol formation . In this report , we demonstrated that calmodulin ( P62158 ) co-immunoprecipitates with the 5- Q13049 receptor in NIH-3T3 fibroblasts in an agonist-dependent manner and that the receptor contains two putative P62158 binding regions . The putative P62158 binding regions of the 5- Q13049 receptor are localized to the second intracellular loop and carboxyl terminus . In an in vitro binding assay peptides encompassing the putative second intracellular loop ( i2 ) and carboxyl-terminal ( ct ) P62158 binding regions bound P62158 in a Ca2+-dependent manner . The i2 peptide bound with apparent higher affinity and shifted the mobility of P62158 in a nondenaturing gel shift assay . Fluorescence emission spectral analyses of dansyl- P62158 showed apparent K(D) values of 65 +/- 30 nM for the i2 peptide and 168 +/- 38 nM for the ct peptide . The ct P62158 -binding domain overlaps with a putative protein kinase C ( PKC ) site , which was readily phosphorylated by PKC in vitro . P62158 binding and phosphorylation of the ct peptide were found to be antagonistic , suggesting a putative role for P62158 in the regulation of 5- Q13049 receptor phosphorylation and desensitization . Finally , we showed that P62158 decreases 5- Q13049 receptor-mediated [35S]GTPgammaS binding to NIH-3T3 cell membranes , supporting a possible role for P62158 in regulating receptor-G protein coupling . These data indicate that the serotonin 5- Q13049 receptor contains two high affinity P62158 -binding domains that may play important roles in signaling and function . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . [ Clinical efficacy of zolmitriptan in migraine ] . Zolmitriptan ( previously known as DB00315 ) is a serotoninergic P28222 /D agonist with high oral bioavailability with a double , central and peripheral , action mechanism . Evaluation of its clinical efficacy was developed in a program of clinical studies ( search and confirmation of dosis , comparative and long term studies ) and through analysis of efficacy in different clinical situations . Zolmitriptan shows a high effectiveness in the treatment of migraine crisis , significantly reduces the headaches by 2 hours of its administration , reduce the symptoms associated with migraine ( nausea , photophobia and phonophobia ) and improves the quality of life of the migraine patient . The efficacy is independent of the type of migraine characteristics of the patient as well as of the administration of other concomitant medications . The dosis of 2.5 mg of zolmitriptan has been found to be the optimum considering both efficacy and tolerability . Serotonergic modulation of the acoustic startle response in rats during preweaning development . The involvement of serotonin ( 5-HT ) in modulating the acoustic startle response ( ASR ) is well established in adult rats , but 5-HT involvement during the preweaning period , when 5-HT neurons undergo extensive development , has not previously been described . Three 5-HT receptor subtypes are reported to modulate the ASR in adult rats : P08908 and 5-HT2 receptor agonists facilitate the ASR , whereas P28222 agonists decrease the response . In the present study , the effects of 5-HT agonists and generalized 5-HT depletion on the ASR were studied in preweanling animals , using independent groups of Long-Evans rats tested on postnatal day ( P01160 ) 13 , 17 and 21 . 8-Hydroxy-2-(di-n-propylamino) tetralin ( 8OHDPAT , 62-1000 micrograms/kg ) , a P08908 receptor agonist , and 5-methoxy-N,N-dimethyl tryptamine ( MeODMT , 2-4 mg/kg ) , a nonselective 5-HT agonist , had no effect on P01160 13 and then increased the ASR on P01160 17 and 21 . The 5-HT2 receptor antagonists cyproheptadine ( 5 mg/kg ) and ketanserin ( 5 mg/kg ) blocked the effect of MeODMT at both ages , providing some evidence that MeODMT increased the ASR through 5-HT2 receptors . 1-(m-Chlorophenyl) piperazine ( mCPP , 1-5 mg/kg ) , a P28222 agonist , had no effect on ASR amplitude on P01160 13 or 17 and then produced a dose-related decrease in the response on P01160 21 . Generalized depletion of 5-HT by 80-90 % in whole-brain and spinal cord , using p-chlorophenylalanine ( PCPA , 300 mg/kg 24 hr prior to testing ) , did not alter ASR amplitude at any age. ( ABSTRACT TRUNCATED AT 250 WORDS ) DB01259 -mediated cyclooxygenase-2 expression via epidermal growth factor receptor/ Q15717 interaction enhances the aggressiveness of triple-negative breast cancer cells . DB01259 , a dual epidermal growth factor receptor ( P00533 ) /human epidermal growth factor receptor 2 ( P04626 ) kinase inhibitor , showed clinical benefits in advanced P04626 -positive breast cancer patients . Because some triple-negative breast cancers ( TNBCs ) frequently overexpress P00533 , the antitumor activity of lapatinib in such diseases was also tested . However , the results showed a worse event-free survival rate . It remains unknown whether and how lapatinib elicits the aggressiveness of such cancer cells . In this study , our results demonstrated that lapatinib facilitated axillary and lung metastases of triple-negative MDA-MB-231 breast cancer cells without affecting their viability , leading to worse survival in orthotopic xenograft mice . The lapatinib-increased motility was attributed by the elevation of P00533 through the downregulation of microRNA-7 and by the subsequent overexpression of cyclooxygenase-2 ( P35354 ) . Strikingly , independent of its kinase activity , the elevated P00533 at least partly stabilized P35354 expression by enhancing the binding of Q15717 to P35354 mRNA . Our results suggest that lapatinib may increase the migration and invasion of MDA-MB-231 cells by upregulating P00533 and P35354 through the downregulation of microRNA-7 , providing a potential explanation for the worse clinical outcome of TNBC patients who receive lapatinib-based treatment . These findings also shed new light on the molecular mechanism of P35354 mRNA stabilization by P00533 in a kinase-independent manner . Effects of serotonin on expression of the P01130 family member Q92673 and 7-ketocholesterol-induced apoptosis in human vascular smooth muscle cells . Serotonin ( 5-HT ) is a known mitogen for vascular smooth muscle cells ( VSMCs ) . The dedifferentiation and proliferation/apoptosis of VSMCs in the arterial intima represent one of the atherosclerotic changes . Q92673 , a member of low-density lipoprotein receptor family , may contribute to the proliferation of VSMCs in neointimal hyperplasia . We conducted an in vitro study to investigate whether 5-HT is involved in Q92673 expression in human VSMCs and apoptosis of VSMCs induced by 7-ketocholesterol ( 7KCHO ) , an oxysterol that destabilizes plaque . 5-HT enhanced the proliferation of VSMCs , and this effect was abolished by sarpogrelate , a selective 5- Q13049 receptor antagonist . Sarpogrelate also inhibited the 5-HT-enhanced Q92673 mRNA expression in VSMCs . Furthermore , 5-HT suppressed the 7KCHO-induced apoptosis of VSMCs via caspase-3/7-dependent pathway . These findings provide new insights on the changes in the differentiation stage of VSMCs mediated by 5-HT . Cortical spreading depression-associated hyperemia in rats : involvement of serotonin . We investigated whether the vasoactive neurotransmitter serotonin ( 5-HT ) is involved in cortical spreading depression ( Q9Y600 ) -associated hyperemia in the rat . We focused on the 5-HT2 receptor , which is engaged in 5-HT induced small arteriolar relaxation in cats , as well as on the P28221 /1B receptor , the binding site of the potent antimigraine drug sumatriptan . In male barbiturate anaesthetized Wistar rats ( n=25 ) CSDs were elicited by brain topical application of 1 M DB00761 , and the DC-potential and regional cerebral blood flow ( rCBF , by Laser Doppler flowmetry ) were measured over the same hemisphere through dura and thinned bone , respectively . Intravenous application of 8 mg/kg of the 5- Q13049 /2C receptor antagonist ritanserin ( group I ; n=8 ) significantly reduced the hyperperfusion amplitude during Q9Y600 by approximately 44 % ( p < 0.05 , from 342+/-124 to 194+/-97 % , baseline before Q9Y600 =100 % ) , and prolonged its duration by approx . 30 % . Vehicle alone ( group II ; n=4 ) did not affect Q9Y600 hyperperfusion . The highly selective P28221 /1B receptor agonist DB00315 was given in two doses : 100 micrograms/kg i.v. ( n=5 ) had no effect on Q9Y600 hyperperfusion , while 800 micrograms/kg ( n=5 ) increased hyperperfusion significantly ( p < 0.05 , from 224+/-86 to 310+/-148 % ) . We conclude that serotonin is , probably via 5-HT2 receptors , involved in the modulation of the regional cerebral blood flow increase during Q9Y600 . Novel highly selective receptor antagonists may help to discriminate the differential contribution of various 5-HT receptor subspecies . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Fluorescence energy transfer analysis of calmodulin-peptide complexes . The interactions between calmodulin and the tryptophan residues of synthetic peptides corresponding to the calmodulin binding domains of skeletal muscle myosin light-chain kinase and the plasma membrane calcium pump were examined . The single tryptophan residue contained in each peptide became relatively immobilized and inaccessible to iodide ion upon binding to calmodulin , indicating that the indole side chain was inserted into a hydrophobic cleft in the surface of calmodulin . Fluorescence energy transfer from peptidyl tryptophan residues to an AEDANS moiety attached to cysteine-26 of spinach calmodulin was measured . Included in these analyses was a tryptophan-containing peptide analog of the calmodulin binding domain of neuromodulin . These data indicated that the indole ring of each peptide inserted 32-35 A away from cysteine-26 and may therefore interact with the carboxyl-terminal lobe of P62158 in its " bent " conformation [ Persechini & Kretsinger ( 1988a ) J. Cardiovasc. Pharmacol. 12 ( Suppl 5 ) , S1- P28222 ; Ikura et al. ( 1992 ) Science 256 , 632-638 ; Vorherr et al. ( 1992 ) Eur. J. Biochem. 204 , 931-937 ] . The interchange of tryptophan-3 and phenylalanine-21 of the calcium pump peptide increased the efficiency of energy transfer to the AEDANS-moiety approximately 12-fold , reducing the calculated distance to 20 A . These data suggest that phenylalanine-21 of the calcium pump peptide interacts with the hydrophobic cleft in the amino-terminal lobe of P62158 . Modulation of multiple sclerosis by sunlight exposure : role of cis-urocanic acid . The role of cis-urocanic acid ( UCA ) as a UV-mediated immunomodulator in MS patients was investigated . Plasma levels of cis-UCA were significantly lower in MS patients compared to controls . Stimulation of MBP- and Q16653 -specific T cells in the presence of cis-UCA , significantly increased P22301 , and inhibited IFN-γ production . PBMCs cultured in the presence of cis-UCA increased P01730 (+)CD25(+)FoxP3(+) regulatory T cell percentages . Dendritic cells cultured in the presence of cis-UCA significantly reduced Ag presentation capacity . Finally , cis-UCA activated the 5- Q13049 receptor , inducing the increase in phosphorylated forms of P29323 1/2 and P45984 . Thus , in addition to vitamin D , cis-UCA also appears to be an additional UV-mediated immunomodulator . Serotonin transporter interacts with the PDGFβ receptor in DB00102 -induced signaling and mitogenesis in pulmonary artery smooth muscle cells . The serotonin transporter ( P31645 ) and the platelet-derived growth factor receptor ( P09619 ) have been implicated in both clinical and experimental pulmonary hypertension ( PH ) and the facilitation of pulmonary artery smooth muscle cell ( PASMC ) growth . To gain a better understanding of the possible relationship of these two cell surface molecules we have explored interactions between P31645 and P09619 . We have previously demonstrated that P31645 transactivates PDGFRβ in serotonin-stimulated PASMC proliferation . We now provide evidence for a role for P31645 in DB00102 signaling and PASMC proliferation by using pharmacological inhibitors , genetic ablation , and construct overexpression of P31645 . The results show that four tested P31645 blockers dose dependently inhibit PDGF-stimulated human and bovine PASMC proliferation with comparable efficacy to that of P09619 inhibitors , whereas P28222 or 5- Q13049 receptor inhibitors had no effect . Combinations of the P31645 and P09619 inhibitors led to synergistic/additive inhibition . Similarly , PDGF-induced PASMC proliferation was attenuated by small interfering RNA downregulation of P31645 . Inhibition of P31645 in PASMCs attenuated PDGF-induced phosphorylation of PDGFRβ , Akt , and p38 but not Erk . Overexpression of P31645 in HEK293 cells led to enhanced Akt phosphorylation by PDGF , which was blunted by a P31645 PDZ motif mutant , indicating the mechanistic need for the PDZ motif of P31645 in PDGF signaling . Furthermore , coimmunoprecipitation experiments showed that P31645 and PDGFRβ become physically associated upon PDGF stimulation . In total , the data show for the first time an important interactive relationship between P31645 and the PDGFRβ in the production of PASMC proliferation triggered by PDGF that may be important in PH .

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O60674 - P40763 blockade by AG490 suppresses autoimmune arthritis in mice via reciprocal regulation of regulatory T Cells and Th17 cells . P05231 -mediated P40763 signaling is essential for Th17 differentiation and plays a central role in the pathogenesis of rheumatoid arthritis . To investigate the molecular mechanism underlying the antirheumatic effects and T cell regulatory effects of P40763 inhibition , we studied the effects of the JAK 2 inhibitor AG490 on Th17 cell/regulatory T cell ( Treg ) balance and osteoclastogenesis . AG490 was administered to mice with collagen-induced arthritis ( CIA ) via i.p. injection , and its in vivo effects were determined . Differential expression of proinflammatory cytokines , including Q16552 , IL-1β , and P05231 , was analyzed by immunohistochemistry . Levels of phosphorylated P40763 and P42229 and differentiation of Th17 cells and Tregs after AG490 treatment in our CIA model were analyzed by immunostaining . In vitro development of Th17 cells and Tregs was analyzed by flow cytometry and real-time PCR . AG490 ameliorated the arthritic phenotype in CIA and increased the proportion of Foxp3(+) Tregs . In contrast , the proportion of Q16552 -producing T cells and levels of inflammatory markers were reduced in AG490-treated mice . Numbers of p- P40763 (+) P01730 (+) T cells and p- P42229 (+) P01730 (+) T cells were reduced and elevated , respectively , after treatment with AG490 . Furthermore , AG490 markedly increased the expression of molecules associated with Treg development ( Q9Y6W8 , programmed cell death protein 1 , P05362 , and CD103 ) . The development and function of osteoclasts were suppressed by AG490 treatment . Our results suggest that AG490 , specifically regulating the O60674 / P40763 pathway , may be a promising treatment for rheumatoid arthritis . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Effects of the total saponins from Rosa laevigata Michx fruit against acetaminophen-induced liver damage in mice via induction of autophagy and suppression of inflammation and apoptosis . The effect of the total saponins from Rosa laevigata Michx fruit ( RLTS ) against acetaminophen ( DB00316 ) -induced liver damage in mice was evaluated in the present paper . The results showed that RLTS markedly improved the levels of liver SOD , CAT , DB00143 , DB00143 -Px , MDA , NO and P35228 , and the activities of serum ALT and Q9NRA2 caused by DB00316 . Further research confirmed that RLTS prevented fragmentation of DNA and mitochondrial ultrastructural alterations based on TdT-mediated dUTP nick end labeling ( TUNEL ) and transmission electron microscopy ( TEM ) assays . In addition , RLTS decreased the gene or protein expressions of cytochrome P450 ( P05181 ) , pro-inflammatory mediators ( IL-1β , P05112 , P05231 , P01375 -α , P35228 , Bax , HMGB-1 and P35354 ) , pro-inflammatory transcription factors ( NF-κB and AP-1 ) , pro-apoptotic proteins ( cytochrome C , p53 , caspase-3 , caspase-9 , p-JNK , p-p38 and p- P29323 ) , and increased the protein expressions of Bcl-2 and Bcl-xL . Moreover , the gene expression of P22301 , and the proteins including LC3 , Q14457 and Atg5 induced by DB00316 were even more augmented by the extract . These results demonstrate that RLTS has hepatoprotective effects through antioxidative action , induction of autophagy , and suppression of inflammation and apoptosis , and could be developed as a potential candidate to treat DB00316 -induced liver damage in the future . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Primary B cell immunodeficiencies : comparisons and contrasts . Sophisticated genetic tools have made possible the identification of the genes responsible for most well-described immunodeficiencies in the past 15 years . Mutations in Btk , components of the pre-B cell and B cell receptor ( lambda5 , Igalpha , Igbeta ) , or the scaffold protein Q8WV28 account for approximately 90 % of patients with defects in early B cell development . Hyper-IgM syndromes result from mutations in P29965 , P25942 , Q9GZX7 , or P13051 in 70-80 % of affected patients . Rare defects in Q9Y6W8 or P15391 can result in a clinical picture that is consistent with common variable immunodeficiency , and as many as 10 % of patients with this disorder have heterozygous amino acid substitutions in O14836 . For all these disorders , there is considerable clinical heterogeneity in patients with the same mutation . Identifying the genetic and environmental factors that influence the clinical phenotype may enhance patient care and our understanding of normal B cell development . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . High biochemical selectivity of tadalafil , sildenafil and vardenafil for human phosphodiesterase 5A1 ( O76074 ) over PDE11A4 suggests the absence of PDE11A4 cross-reaction in patients . The physiological role of phosphodiesterase (PDE)11 is unknown and its biochemical characteristics are poorly understood . We have expressed human DB00117 -tagged PDE11A4 and purified the enzyme to apparent homogeneity . PDE11A4 displays K(m) values of 0.97 microM for cGMP and 2.4 microM for DB02527 , and maximal velocities were 4- to 10-fold higher for DB02527 than for cGMP . Given the homology between PDE11 and O76074 , we have compared the biochemical potencies of tadalafil ( DB00820 , Lilly- Q9Y6W8 ) , vardenafil ( DB00862 , Bayer-GSK ) , and sildenafil ( Viagra , Pfizer Inc. ) for PDE11A4 and PDE5A1 . PDE5A1/PDE11A4 selectivities are 40- , 9300- , and 1000-fold for tadalafil , vardenafil , and sildenafil , respectively . This suggests that none of these three compounds is likely to crossreact with PDE11A4 in patients . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Emerging oral drugs for erectile dysfunction . Erectile dysfunction ( ED ) is a common medical condition that affects the sexual life of millions of men worldwide . Many drugs are now available for the treatment of ED , with oral pharmacotherapy representing the first-line option for most patients . DB00203 citrate , an inhibitor of the enzyme phosphodiesterase type 5 ( O76074 ) , is the most widely prescribed oral agent and has a very satisfactory efficacy-safety profile in all patient categories . DB00820 ( DB00820 ; Eli Lilly & Co. , Q9Y6W8 ) and vardenafil ( DB00862 ; Bayer Pharmaceuticals , GlaxoSmithKline ) are new O76074 inhibitors that have recently been approved worldwide . Both have been associated with significant positive efficacy-safety profiles . DB00714 sublingual is a dopamine D1 and D2 receptor agonist , which has been approved for marketing in Europe . It is best selected for treating patients with mild-to-moderate ED , but it is seldom used in clinical practice due to its limited efficacy and side effects , particularly nausea . Patients who do not respond to oral pharmacotherapy or who are unable to use it are appropriate candidates for intracavernosal and intraurethral therapy . The efficacy of second-line treatment is high , but the attrition rate remains significant . For the purpose of this review , clinical and pharmacological analysis focuses on the recent advances in the field of oral therapy , including O76074 inhibitors and sublingual apomorphine . Brain 5-HT and inhibition of aggressive behavior in animals : 5-HIAA and receptor subtypes . Evolutionary constant serotonin ( 5-HT ) neuronal systems evolved along medial brain structures ; yet , wide variations in functionality characterize serotonergic systems in mediating aggressive responses in species ranging from lobsters , ants , electric fish , and rodents to primates . So far , the attempts to correlate cerebrospinal fluid ( P04141 ) 5-hydroxyindoleacetic acid ( 5-HIAA ) levels with measures of aggression have revealed inverse , direct , or no correlations in different nonhuman primate species . It is difficult to harmonize the occasional correlations between P04141 5-HIAA and adaptive aggressive acts in nonhuman primates ( a ) with clinically diagnosed suicidal or impulsive individuals , and ( b ) with the biochemical , anatomical , and presumably functional differentiation of 5-HT pathways and receptor subtypes . Eltoprazine , a mixed P08908 /B agonist , and meta-trifluoro-methylphenyl-piperazine HCl ( TFMPP ) , a more selective P28222 agonist , specifically decrease aggressive behavior in several animal species and situations in both sexes without detriment to other social , exploratory , or motoric activities . A definite role for P08908 , 5-HT2 , and 5- Q9H205 receptor subtypes in the mechanisms mediating aggressive behaviors has to await the development of selective agonists and antagonists , respectively . Induction , binding specificity and function of human Q9Y6W8 . Recently , we have identified the inducible co-stimulator ( Q9Y6W8 ) , an activation-dependent , T cell-specific cell surface molecule related to P10747 and P16410 . Detailed analysis of human Q9Y6W8 presented here shows that it is a 55-60-kDa homodimer with differently N-glycosylated subunits of 27 and 29 kDa . Q9Y6W8 requires both phorbol 12-myristate 13-acetate and ionomycin for full induction , and is sensitive to DB00091 . Q9Y6W8 is up-regulated early on all T cells , including the P10747 - subset , and continues to be expressed into later phases of T cell activation . On stimulation of T cells by antigen-presenting cells , the P10747 / P33681 , but not the P29965 / P25942 pathway is critically involved in the induction of Q9Y6W8 . Q9Y6W8 does not bind to P33681 -1 or P33681 -2 , and P10747 does not bind to O75144 ; thus the P10747 and Q9Y6W8 pathways do not cross-interact on the cell surface . In vivo , Q9Y6W8 is expressed in the medulla of the fetal and newborn thymus , in the T cell zones of tonsils and lymph nodes , and in the apical light zones of germinal centers ( predominant expression ) . Functionally , Q9Y6W8 co-induces a variety of cytokines including P05112 , P05113 , P05231 , P01579 , P01375 , GM- P04141 , but not P60568 , and superinduces P22301 . Furthermore , Q9Y6W8 co-stimulation prevents the apoptosis of pre-activated T cells . The human Q9Y6W8 gene maps to chromosome 2q33 - 34 . P10747 and P16410 coreceptor expression and signal transduction . T-cell activation is mediated by antigen-specific signals from the TCRzeta/CD3 and P01730 -CD8-p56lck complexes in combination with additional co-signals provided by coreceptors such as P10747 , inducible costimulator ( Q9Y6W8 ) , cytotoxic T-lymphocyte antigen-4 ( P16410 ) , programmed death ( P18621 ) , and others . P10747 and Q9Y6W8 provide positive signals that promote and sustain T-cell responses , while P16410 and P18621 limit responses . The balance between stimulatory and inhibitory co-signals determines the ultimate nature of T-cell responses where response to foreign pathogen is achieved without excess inflammation and autoimmunity . In this review , we outline the current knowledge of the P10747 and P16410 signaling mechanisms [ involving phosphatidylinositol 3 kinase ( PI3K ) , growth factor receptor-bound protein 2 ( Grb2 ) , Filamin A , protein kinase C theta ( PKCtheta ) , and phosphatases ] that control T-cell immunity . We also present recent findings on Q6PIZ9 ( Q6PIZ9 ) regulation of P16410 surface expression , and a signaling pathway involving P16410 activation of PI3K and protein kinase B ( P31749 ) /AKT by which cell survival is ensured under conditions of anergy induction . B cells alter the phenotype and function of follicular-homing P32302 + T cells . The CXC chemokine receptor (CXCR)5 is rapidly induced on activated P01730 (+) T cells , allowing migration toward secondary lymphoid tissue follicles , where the P32302 ligand O43927 / O43927 is produced . Such P32302 (+) T cells provide efficient help for B cell immunoglobulin production and are termed follicular B helper T ( T(FH) ) cells . However , the molecular mechanisms by which T(FH) cells provide B cell help are unknown . Here , we demonstrate that newly generated ( antigen-primed ) T(FH) cells express a phenotype consistent with induction of B cell proliferation , but co-culture with primed B cells resulted in a switch to a plasma cell-inducing phenotype , characterized by loss of CD154 , induction of P32970 and an increase in P22301 production capacity . The ability to produce P22301 could be maintained as a stable phenotype , but its secretion was strictly dependent on inducible costimulator ( Q9Y6W8 ) signaling . Furthermore , B cells preserved a lymph node migration phenotype in proliferating T(FH) cells by preventing the loss of CC chemokine receptor (CCR)7 and the induction of P51681 . Thus , B cells directly modulate the B cell helper phenotype in T(FH) cells and actively promote their prolonged co-localization with these cells . Pituitary-adrenal axis regulation in P06850 -deficient mice . P06850 ( P06850 ) -deficient ( knockout ( KO ) ) mice demonstrate severely impaired adrenal responses to restraint , ether , and fasting , and lack the normal diurnal glucocorticoid ( GC ) rhythm . Here , we summarize recent studies determining the role of P06850 in augmenting plasma adrenocorticotrophic hormone ( DB01285 ) concentration after glucocorticoid withdrawal and pituitary-adrenal axis stimulation in the context of inflammation . Even though GC insufficient , basal pituitary proopiomelanocortin ( P01189 ) mRNA , DB01285 peptide content within the pituitary , and plasma DB01285 concentrations are not elevated in P06850 KO mice . P01189 mRNA content in P06850 KO mice increases following adrenalectomy , and this increase is reversed by GC , but not aldosterone , replacement . In marked contrast to the increase in P01189 mRNA , plasma DB01285 does not increase in the P06850 KO mice following adrenalectomy . Administration of P06850 to adrenalectomized P06850 KO mice results in acute , robust DB01285 secretion . Thus , loss of GC feedback can increase P01189 gene expression in the pituitary , but P06850 action is essential for increased secretion of DB01285 into the circulation . While GC secretion is impaired in P06850 KO mice after most stimuli , we have found near-normal GC responses to inflammation and systemic immune challenge . Studies in mice with P06850 and P05231 deficiency reveal that P05231 is essential for activation of the pituitary-adrenal axis during inflammatory and other stressors in the absence of P06850 . Southwest Oncology Group study S0413 : a phase II trial of lapatinib ( GW572016 ) as first-line therapy in patients with advanced or metastatic gastric cancer . BACKGROUND : DB01259 ( GW572016 ) is a dual tyrosine kinase inhibitor of epidermal growth factor receptor ( P00533 ) and human epidermal growth factor receptor 2 ( P04626 /ErbB2 ) , which are reported as overexpressed in 15 % -45 % of gastric cancers , making them potential targets . PATIENTS AND METHODS : The primary objective of this study was to assess response rate . Secondary objectives included overall survival ( OS ) , toxicity , and the relationship of P00533 , ErbB2 , and markers of angiogenesis with clinical outcome . DB01259 was administered to chemonaive metastatic gastric cancer patients at a dose of 1500 mg orally daily for 28 days . RESULTS : The study enrolled 47 patients from February 2005 until May 2006 . Four patients ( 9 % ) had a confirmed partial response ( PR ) , 1 ( 2 % ) had an unconfirmed PR , and 10 ( 23 % ) had stable disease . Median ( 95 % confidence interval ) time to treatment failure was 1.9 ( 1.6-3.1 ) months and OS was 4.8 ( 3.2-7.4 ) months . Significant adverse events : one grade 4 cardiac ischemia/infarction , one grade 4 fatigue , and one grade 4 emesis . One treatment-related death was due to central nervous system ischemia . An exploratory analysis of markers revealed gene expression of P04626 , interleukin ( IL ) -8 and genomic polymorphisms P10145 , and vascular endothelial growth factor correlated with OS . CONCLUSIONS : DB01259 is well tolerated , with modest single-agent activity in advanced/metastatic gastric cancer patients . Potential molecular correlatives were identified which warrant further validation . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . Radiolabeled ligand binding to the catalytic or allosteric sites of O76074 and PDE11 . Cyclic nucleotide phosphodiesterases ( PDEs ) have been investigated for years as targets for therapeutic intervention in a number of pathophysiological processes . Phosphodiesterase-5 ( O76074 ) , which is highly specific for guanosine 3'-5'-cyclic-monophosphate ( cGMP ) at both its catalytic site and its allosteric sites , has generated particular interest because it is potently and specifically inhibited by three drugs : sildenafil ( Viagra , Pfizer ) , tadalafil ( DB00820 , Lilly- Q9Y6W8 ) , and vardenafil ( DB00862 , Bayer GSK ) . Previously , we have used [(3)H]cGMP to directly study the interaction of cGMP with the allosteric sites of O76074 , but because cGMP binds with relatively low affinity to the catalytic site , it has been difficult to devise a binding assay for this particular binding reaction . This approach using measurement of radiolabeled ligand binding continues to allow us to more precisely define functional features of the enzyme . We now use a similar approach to study the characteristics of high-affinity [(3)H]inhibitor binding to the O76074 catalytic domain . For these studies , we have prepared [(3)H]sildenafil and [(3)H]tadalafil , two structurally different competitive inhibitors of O76074 . The results demonstrate that radiolabeled ligands can be used as probes for both catalytic site and allosteric site functions of O76074 . We describe herein the methods that we have established for studying the binding of radiolabeled ligands to both types of sites on O76074 . These techniques have also been successfully applied to the study of binding of radiolabeled O76074 inhibitors to PDE11 , suggesting that these methods are applicable to the study of other PDEs , and perhaps other enzyme families . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Expression and function of the inducible costimulator ligand O75144 in human airway smooth muscle cells . BACKGROUND : O75144 is a ligand for the inducible costimulator ( Q9Y6W8 ) . The aim of this study was to examine the expression and function of O75144 in human airway smooth muscle ( P17405 ) cells and compare them with those of P25942 or P23510 ( P23510 ) . METHODS : Expression of O75144 , P25942 and P23510 in P17405 cells and their respective counterparts in T cells was analyzed by RT-PCR or flow cytometry . The modulating effect of polyinosinic-polycytidylic acid ( poly I:C ) on expression of O75144 , P25942 and P23510 was also examined . The function of these three molecules was evaluated by virtue of adhesion of anti-CD3-activated T cells , P05231 and P10145 production and DNA synthesis . RESULTS : P17405 cells constitutively expressed O75144 , P25942 and P23510 that mediated adhesion of activated T cells expressing Q9Y6W8 , P29965 and OX40 . P17405 cells responded to poly I:C with upregulated expression of O75144 , P25942 and P23510 and displayed enhanced adhesion of activated T cells . Functional analysis performed on untreated P17405 cells showed that engagement of O75144 with Q9Y6W8 -Ig clearly induced DNA synthesis , whereas that of P25942 or P23510 with trimeric P29965 or OX40-Ig greatly increased P05231 and P10145 production . These responses were enhanced in poly I:C-treated P17405 cells . CONCLUSIONS : The data demonstrate that P17405 cells express functionally active O75144 , P25942 and P23510 and suggest that O75144 -dependent signaling may play an active role in a proliferative response rather than in cytokine and chemokine production . In addition , the modulation of O75144 , P25942 and P23510 expression and function by poly I:C may have important implications for the function of virus-infected P17405 cells . Murine lupus susceptibility locus Sle1a controls regulatory T cell number and function through multiple mechanisms . The Sle1 locus is a key determinant of lupus susceptibility in the NZM2410 mouse model . Within Sle1 , we have previously shown that Sle1a expression enhances activation levels and effector functions of P01730 (+) T cells and reduces the size of the P01730 (+)CD25(+)Foxp3(+) regulatory T cell subset , leading to the production of autoreactive T cells that provide help to chromatin-specific B cells . In this study , we show that Sle1a P01730 (+) T cells express high levels of Q9Y6W8 , which is consistent with their increased ability to help autoreactive B cells . Furthermore , Sle1a P01730 (+)CD25(+) T cells express low levels of Foxp3 . Mixed bone marrow chimeras demonstrated that these phenotypes require Sle1a to be expressed in the affected P01730 (+) T cells . Expression of other markers generally associated with regulatory T cells ( Tregs ) was similar regardless of Sle1a expression in Foxp3(+) cells . This result , along with in vitro and in vivo suppression studies , suggests that Sle1a controls the number of Tregs rather than their function on a per cell basis . Both in vitro and in vivo suppression assays also showed that Sle1a expression induced effector T cells to be resistant to Treg suppression , as well as dendritic cells to overproduce P05231 , which inhibits Treg suppression . Overall , these results show that Sle1a controls both Treg number and function by multiple mechanisms , directly on the Tregs themselves and indirectly through the response of effector T cells and the regulatory role of dendritic cells .

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DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Effects of eletriptan on the peptidergic innervation of the cerebral dura mater and trigeminal ganglion , and on the expression of c-fos and c-jun in the trigeminal complex of the rat in an experimental migraine model . Nociceptive axons and terminals in the supratentorial cerebral dura mater display an intense calcitonin gene-related peptide ( P80511 ) immunoreactivity . In an experimental migraine model , it has been shown that electrical stimulation of the rat trigeminal ganglion induced an increase in the lengths of P80511 -immunoreactive axons , increased size and number of pleomorphic axonal varicosities in the dura mater , and an increased number of c-jun and c-fos protein-expressing nerve cells in the trigeminal complex . We demonstrate the effect of the highly specific and moderately lipophilic serotonin agonist eletriptan ( Pfizer ) which prevents the effects of electrical stimulation in the dura mater . DB00216 also affected the caudal trigeminal complex ; it markedly reduced the numbers of the oncoprotein-expressing cells , mainly after stimulation and to some extent also in nonstimulated animals . DB00216 also affected expression of P80511 in perikarya of trigeminal ganglion cells , insofar as the number of small nerve cells exhibiting a compact P80511 immunoreaction was decreased to one quarter of the original value . In all these respects , eletriptan acted in a similar way to sumatriptan , with the notable exception that eletriptan also blocked the stimulation-induced effects in the nucleus caudalis trigemini and the upper cervical spinal cord ( trigeminal complex ) , whereas sumatriptan did not . It is concluded that eletriptan , acting on perikarya and both the peripheral and the central axon terminals of primary sensory neurons , exerts its antimigraine effect by an agonist action on P28222 /1D receptors throughout the entire trigeminal system , probably by passing the blood-brain-barrier because of its lipophilic character . Characterisation of the 5-HT receptor binding profile of eletriptan and kinetics of [3H]eletriptan binding at human P28222 and P28221 receptors . The affinity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] -1H-indole ) for a range of 5-HT receptors was compared to values obtained for other P28222 /1D receptor agonists known to be effective in the treatment of migraine . DB00216 , like sumatriptan , zolmitriptan , naratriptan and rizatriptan had highest affinity for the human P28222 , P28221 and putative 5-ht1f receptor . Kinetic studies comparing the binding of [3H]eletriptan and [3H]sumatriptan to the human recombinant P28222 and P28221 receptors expressed in HeLa cells revealed that both radioligands bound with high specificity ( > 90 % ) and reached equilibrium within 10-15 min . However , [3H]eletriptan had over 6-fold higher affinity than [3H]sumatriptan at the P28221 receptor ( K(D) ) : 0.92 and 6.58 nM , respectively ) and over 3-fold higher affinity than [3H]sumatriptan at the P28222 receptor ( K(D) : 3.14 and 11.07 nM , respectively ) . Association and dissociation rates for both radioligands could only be accurately determined at the P28221 receptor and then only at 4 degrees C . At this temperature , [3H]eletriptan had a significantly ( P < 0.05 ) faster association rate ( K(on) 0.249 min(-1) nM(-1) ) than [3H]sumatriptan ( K(on) 0.024 min(-1) nM(-1) ) and a significantly ( P < 0.05 ) slower off-rate ( K(off) 0.027 min(-1) compared to 0.037 min(-1) for [3H]sumatriptan ) . These data indicate that eletriptan is a potent ligand at the human P28222 , P28221 , and 5-ht1f receptors and are consistent with its potent vasoconstrictor activity and use as a drug for the acute treatment of migraine headache . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Activation of Akt1 by human 5-hydroxytryptamine (serotonin)1B receptors is sensitive to inhibitors of MEK . Akt1/protein kinase B and the mitogen-activated protein ( Q96HU1 ) kinases extracellular signal-regulated kinase 1 ( P27361 ) and P28482 have been shown to promote cell survival in a cell-specific manner . Since many receptors activate both pathways , inhibitors are commonly used to study the relative role of each pathway . In the present study , we examined the effects of PD098059 and U0126 , two structurally dissimilar inhibitors of Q96HU1 kinase kinase ( Q02750 /2 ) , on the activation of P29323 and Akt stimulated by human 5-hydroxytryptamine(1B) ( serotonin ) ( P28222 ) receptors . Surprisingly , pathways for activation of both P29323 and Akt were found to be sensitive to the two MEK inhibitors at concentrations commonly used to selectively inhibit the activation of P29323 . Both compounds caused complete inhibition of phosphorylation of P29323 and a maximal 60 % inhibition of P28222 receptor-mediated phosphorylation of Akt . Inhibition of Akt activation required almost complete inhibition of P29323 . Transfection with cDNA for activated forms of Q02750 /2 caused increased phosphorylation of P29323 but not of Akt , demonstrating that independent activation of MEK/ P29323 was insufficient for activation of Akt . Therefore , it is not clear whether inhibition of activation of Akt resulted from selective inhibition of MEK or from additional actions on other unidentified common pathways . Nevertheless , our findings that PD098059 and U0126 inhibit activation of Akt at commonly used concentrations demonstrate that in at least some systems , these compounds inhibit activation of both P29323 and Akt , and can not be used to discern the relative roles of each pathway in mediating cellular responses . Characterisation of the contractile activity of eletriptan at the canine vascular P28222 receptor . The functional activity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] - 1 H-indole ) at the contractile serotonin ( 5-hydroxytryptamine ; 5-HT ) ' 1B-like ' receptor in dog isolated saphenous vein and basilar artery was investigated . DB00216 , like 5-HT and sumatriptan potently contracted saphenous vein ( pEC50 : 6.3 , 6.9 and 6.1 , respectively ) and basilar artery ( pEC50 7.2 , 7.5 and 6.8 , respectively ) . The maximum responses evoked by eletriptan was , unlike sumatriptan , significantly lower than that to 5-HT ( intrinsic activity saphenous vein : eletriptan 0.57 , 5-HT 1.0 , sumatriptan 0.85 ; basilar artery : eletriptan 0.77 , 5-HT 0.98 , sumatriptan 0.89 ) . Contractions evoked by eletriptan were antagonised by the P28222 /1D receptor antagonist GR125743 ( N- [ 4-methoxy-3- ( 4-methyl piperazin-1-yl ) phenyl ] -3-methyl-4-(4-pyridyl)benzamide ) with pA2 values of 9.1 in saphenous vein and 9.4 in basilar artery . Affinity estimates ( pKA ) for 5-HT and sumatriptan determined from receptor alkylation studies in saphenous vein were 6.6 and 6.3 , respectively , compared to the apparent equilibrium dissociation constant ( pKp ) for eletriptan of 6.8 . The rank order of relative intrinsic efficacies ( epsilon ) was 5-HT > sumatriptan > eletriptan . Thus , eletriptan required greater receptor occupancy ( 4.4-fold ) to evoke an equivalent contraction to 5-HT and sumatriptan in dog isolated saphenous vein . These data demonstrate that eletriptan is a potent partial agonist at the canine vascular P28222 receptor . P01258 gene-related peptide acts as a mitogen for human Gin-1 gingival fibroblasts by activating the Q96HU1 kinase signalling pathway . In many peripheral tissues , calcitonin gene-related peptide ( P80511 ) is released from peptidergic sensory nerve fibres and acts like a growth factor during tissue development and regeneration . However , the ability of P80511 to influence gingival tissue has not been studied . To address this question , we have now examined the effects of P80511 on the proliferation of human gingival fibroblasts ( Gin-1 ) in vitro . Gin-1 cells have approximately 3100 specific P80511 -binding sites with a Kd of 38.6 pM on their surface . Treatment with P80511 ( 0.1-100 nM ) significantly stimulated cell proliferation in a dose-dependent manner , with maximal effects at 1-10 nM P80511 after 2 d . As one early cellular response to P80511 , P27361 protein ( also known as the extracellular signal response kinase [ P29323 ] ) was tyrosine- and threonine-phosphorylated within 2 min , and this phosphorylation was sustained for at least 1 h . The dose-response curve of MAPK activation was very similar to that observed for P80511 's stimulation of cell proliferation . In addition , P80511 's activation of MAPK stimulated its ability to phosphorylate the Elk-1 transcription factor . When cells were pretreated with PD98059 , a selective inhibitor of MAPK kinase ( also known as MEK ) , P80511 not only failed to induce phosphorylation of MAPK but also failed to stimulate Gin-1 cell proliferation . Our present data indicate that P80511 rapidly activates the MAPK signalling pathway , an effect which consequently stimulates the proliferation of gingival fibroblasts . Our data demonstrate specific cellular responses to P80511 by gingival fibroblasts and support the possibility that P80511 acts as a targeted local factor in the regulation of development , generation and/or regeneration of gingival tissues . Activation of the JAK/ P35610 pathway in vascular smooth muscle by serotonin . Serotonin ( 5-hydroxytryptamine , 5-HT ) is a vasoconstrictor and mitogen whose levels are elevated in diabetes . Previous studies have shown the presence of 5- Q13049 , P41595 , and P28222 receptors in vascular smooth muscle cells ( VSMCs ) . There are currently no data regarding P41595 and P28222 receptor activation of the JAK/ P35610 pathway in VSMCs and resultant potential alterations in 5-HT signaling in diabetes . Therefore , we tested the hypothesis that 5-HT differentially activates the JAK/ P35610 pathway in VSMCs under conditions of normal ( 5 mM ) and high ( 25 mM ) glucose . Treatment of rat VSMCs with 5-HT ( 10(-6) M ) resulted in time-dependent activation ( approximately 2-fold ) of O60674 , P23458 , and P42224 , but not P40763 ( maximal at 5 min , returned to baseline by 30 min ) . The P41595 receptor agonist BW723C86 and the P28222 receptor agonist CGS12066A ( 10(-9)-10(-5) M , 5-min stimulation ) did not activate the JAK/ P35610 pathway . Treatment with the 5- Q13049 receptor antagonist ketanserin ( 10 nM ) inhibited O60674 activation by 5-HT . Treatment of streptozotocin-induced diabetic rats with ketanserin ( 5 mg.kg-1.day-1 ) reduced activation of O60674 and P42224 but not P40763 in endothelium-denuded thoracic aorta in vivo . 5-HT ( 10(-6) M ) treatment resulted in increased cell proliferation and increased DNA synthesis , which were inhibited by the O60674 inhibitor AG490 . Further studies with apocynin , diphenyleneiodonium chloride , catalase , and virally transfected superoxide dismutase had no effect at either glucose concentration on activation of the JAK/ P35610 pathway by 5-HT . Therefore , we conclude that 5-HT activates O60674 , P23458 , and P42224 via the 5- Q13049 receptors in a reactive oxygen species-independent manner under both normal and high glucose conditions . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . DB00216 Pfizer . Pfizer has developed and launched eletriptan , a P28222 /1D agonist , for the potential treatment of migraine with and without aura . DB00216 has 6-fold greater affinity for the P28221 receptor than sumatriptan , and a 3-fold greater affinity for the P28222 receptor [ 249570 ] . DB00216 pharmacology has also been evaluated in vitro in comparison with zolmitriptan ( AstraZeneca plc ) and naratriptan ( GlaxoSmithKline plc ) [ 290116 ] . Porcine carotid vascular effects of eletriptan ( UK-116,044 ) : a new P28222 /1D receptor agonist with anti-migraine activity . It has been suggested that opening of cephalic arteriovenous anastomoses may be involved in the headache phase of migraine . Indeed , a number of acutely acting anti-migraine drugs , including the ergot alkaloids and sumatriptan , constrict porcine carotid arteriovenous anastomoses . In this study , using pentobarbital anaesthetised pigs , we investigated the effects of eletriptan , a close structural analogue of sumatriptan , on the distribution of common carotid artery blood flow into arteriovenous anastomotic and nutrient ( capillary ) fractions . DB00216 ( 10 , 30 , 100 , 300 and 1000 microg kg(-1) , i.v. ) decreased the total carotid blood flow , exclusively by decreasing cephalic arteriovenous anastomotic blood flow ; nutrient blood flow , particularly to the ear , skin and fat , was significantly increased . The doses of eletriptan needed to reduce arteriovenous anastomotic blood flow and conductance by 50 % ( ED50 ) were , respectively , 117+/-21 microg kg(-1) ( 251+/-45 nmol kg(-1) ) and 184+/-42 microg kg(-1) ( 396+/-91 nmol kg(-1) ) ; the highest dose caused reductions of 84+/-3 % and 77+/-4 % , respectively . The eletriptan-induced changes in carotid haemodynamics were clearly attenuated by pretreating the pigs with the selective P28222 /1D receptor antagonist GR127935 ( 0.5 mg kg(-1) ) . On the basis of these results , we conclude that ( 1 ) the eletriptan-induced constriction of cephalic arteriovenous anastomoses as well as the arteriolar dilatation in head tissues is predominantly mediated by P28222 /1D receptors , and ( 2 ) eletriptan should be effective in aborting migraine headache . Clinical studies have already demonstrated its therapeutic action in migraine patients . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . 5-hydroxytryptamine stimulates phosphorylation of Q8TCB0 / Q8NFH3 mitogen-activated protein kinase activation in bovine aortic endothelial cell cultures . 5-Hydroxytryptamine ( 5-HT ) is sequestered and released by endothelial cells , acts as an endothelial cell mitogen , promotes the release of nitric oxide ( NO ) , and has been associated with the Q8TCB0 / Q8NFH3 mitogen-activated protein kinase ( MAPK ) cascade . NO also acts as a cell mitogen and promotes signals that culminate in the phosphorylation of MAPK . The aim of this study was to test whether endothelial 5-HT receptors stimulate dual ( tyrosyl- and threonyl- ) phosphorylation of MAPK through a mitogen-activated protein kinase kinase-1 ( MEK-1 ) and P29474 -dependent pathway in bovine aortic endothelial cells ( BAECs ) . As shown by Western blot analysis , 5-HT and the P28222 -selective agonist 5-nonyloxytryptamine ( 5-NOT ) stimulate time- and concentration-dependent ( 0.001-10 microM ) phosphorylation of MAPK in these cells . The agonist-stimulated phosphorylation of MAPK was blocked by the 5-HT1b-receptor antagonist isamoltane ( 0.01-10 p3M ) and the MEK-1 inhibitor PD 098059 ( [ 2-(2'-amino-3'-methoxy-phenyl)-oxanaphthalen-4-one ] ; 0.01-10 microM¿ . The P29474 inhibitor L-N(omega)-iminoethyl-L-ornithine ( L-NIO ; 0.01-10 microM ) failed to block the 1 microM 5-NOT-stimulated responses . Our findings suggest that the 5-HT receptors ( specifically P28222 ) mediate signals to MEK-1 and subsequently to MAPK through an P29474 -independent pathway in BAECs . A new clinical evidence-based gene-environment interaction model of depression . In our current understanding of mood disorders , the role of genes is diverse including the mediation of the effects of provoking and protective factors . Different or partially overlapping gene sets play a major role in the development of personality traits including also affective temperaments , in the mediation of the effects of environmental factors , and in the interaction of these elements in the development of depression . Certain genes are associated with personality traits and temperaments including e.g. , neuroticism , impulsivity , openness , rumination and extroversion . Environmental factors consist of external ( early and provoking life events , seasonal changes , social support etc. ) and internal factors ( hormones , biological rhythm generators , comorbid disorders etc ) . Some of these environmental factors , such as early life events and some prenatal events directly influence the development of personality traits and temperaments . In the NEWMOOD cohort polymorphisms of the genes of the serotonin transporter , P08908 , P28222 and 5- Q13049 and endocannabinoid P21554 receptors , tryptophan hydroxylase , P16220 , P23560 and GIRK provide evidence for the involvement of these genes in the development of depression . Based on their role in this process they could be assigned to different gene sets . The role of certain genes , such as promoter polymorphisms of the serotonin transporter ( 5-HTTLPR ) and P21554 receptor has been shown in more than one of the above factors . Furthermore , gene-gene interactions of these promoters associated with anxiety suggest the application of these polymorphisms in personalized medicine . In this review we introduce a new model including environmental factors , genes , trait and temperament markers based on human genetic studies . Gastroprotective action of orexin-A against stress-induced gastric damage is mediated by endogenous prostaglandins , sensory afferent neuropeptides and nitric oxide . O43612 -A , identified in the neurons and endocrine cells in the gut , has been implicated in control of food intake and sleep behavior but little is known about its influence on gastric secretion and mucosal integrity . The effects of orexin-A on gastric secretion and gastric lesions induced in rats by 3.5 h of water immersion and restraint stress ( P23381 ) or 75 % ethanol were determined . O43612 -A ( 5-80 microg/kg i.p. ) increased gastric acid secretion and attenuated gastric lesions induced by P23381 and this was accompanied by the significant rise in plasma orexin-A , P80511 and gastrin levels , the gastric mucosal blood flow ( GBF ) , luminal NO concentration and an increase in mRNA for P80511 and overexpression of P35354 protein and the generation of PGE(2) in the gastric mucosa . O43612 -A-induced protection was abolished by selective OX-1 receptor antagonist , vagotomy and attenuated by suppression of P23219 and P35354 , deactivation of afferent nerves with neurotoxic dose of capsaicin , pretreatment with CCK(2)/gastrin antagonist , P80511 (8-37) or capsazepine and by inhibition of NOS with DB04223 . This study shows for the first time that orexin-A exerts a potent protective action on the stomach of rats exposed to non-topical ulcerogens such as P23381 or topical noxious agents such as ethanol and these effects depend upon hyperemia mediated by P36551 -PG and NOS-NO systems , activation of vagal nerves and sensory neuropeptides such as P80511 released from sensory nerves probably triggered by an increase in gastric acid secretion induced by this peptide . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Correcting human mitochondrial mutations with targeted RNA import . Mutations in the human mitochondrial genome are implicated in neuromuscular diseases , metabolic defects , and aging . An efficient and simple mechanism for neutralizing deleterious mitochondrial DNA ( mtDNA ) alterations has unfortunately remained elusive . Here , we report that a 20-ribonucleotide stem-loop sequence from the H1 RNA , the RNA component of the human RNase P enzyme , appended to a nonimported RNA directs the import of the resultant RNA fusion transcript into human mitochondria . The methodology is effective for both noncoding RNAs , such as tRNAs , and mRNAs . The RNA import component , polynucleotide phosphorylase ( Q8TCS8 ) , facilitates transfer of this hybrid RNA into the mitochondrial matrix . In addition , nucleus-encoded mRNAs for mitochondrial proteins , such as the mRNA of human mitochondrial ribosomal protein P28222 ( O15235 ) , contain regulatory sequences in their 3'-untranslated region ( UTR ) that confers localization to the mitochondrial outer membrane , which is postulated to aid in protein translocation after translation . We show that for some mitochondrial-encoded transcripts , such as P35354 , a 3'-UTR localization sequence is not required for mRNA import , whereas for corrective mitochondrial-encoded tRNAs , appending the 3'-UTR localization sequence was essential for efficient fusion-transcript translocation into mitochondria . In vivo , functional defects in mitochondrial RNA ( mtRNA ) translation and cell respiration were reversed in two human disease lines . Thus , this study indicates that a wide range of RNAs can be targeted to mitochondria by appending a targeting sequence that interacts with Q8TCS8 , with or without a mitochondrial localization sequence , providing an exciting , general approach for overcoming mitochondrial genetic disorders . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Investigation of the binding of isoform-selective inhibitors to prostaglandin endoperoxide synthases using fluorescence spectroscopy . Prostaglandin endoperoxide synthase ( PGHS ) is a heme protein that catalyzes the committed step in prostaglandin and thromboxane biosynthesis . Two isoforms of PGHS exist , a constitutive form termed P23219 and an inducible form termed P35354 . We report here fluorescence resonance energy transfer analysis of isoform-selective inhibitors interacting with P23219 and P35354 . By measuring fluorescence quenching due to the energy transfer of the inhibitor fluorescence to the heme prosthetic group of PGHS , we determined these inhibitors bind in the arachidonic acid substrate access channel with an R0 of 35 A for P23219 with the P23219 inhibitor and an R0 of 21 A for P35354 with the P35354 inhibitor . The observed fluorescence quenching is completely dynamic and dominated by quenching by the heme . Time-resolved results combined with molecular modeling determine the distance from the inhibitor to the heme moiety to be 20 A in P23219 and 18 A in P35354 . Preliminary stopped-flow kinetic studies reveal that the rate of quenching is limited by a first-order protein transition , which is slow , and that bound inhibitor undergoes rapid exchange . Functional inhibition related to structure of a highly potent insulin-specific CD8 T cell clone using altered peptide ligands . P01308 -reactive CD8 T cells are amongst the earliest islet-infiltrating CD8 T cells in NOD mice . Cloned insulin B15-23-reactive cells ( designated G9C8 ) , restricted by H-2K(d) , are highly diabetogenic . We used altered peptide ligands ( APL ) substituted at TCR contact sites , positions (p)6 and 8 , to investigate G9C8 T cell function and correlated this with structure . Cytotoxicity and P01579 production assays revealed that p6G and p8R could not be replaced by any naturally occurring amino acid without abrogating recognition and functional response by the G9C8 clone . When tested for antagonist activity with APL differing from the native peptide at either of these positions , the peptide variants , G6H and R8L showed the capacity to reduce the agonist response to the native peptide . The antagonist activity in cytotoxicity and P01579 production assays can be correlated with conformational changes induced by different structures of the MHC-peptide complexes , shown by molecular modeling . We conclude that P80511 and p8 of the insulin B15-23 peptide are very important for TCR stimulation of this clone and no substitutions are tolerated at these positions in the peptide . This is important in considering the therapeutic use of peptides as APL that encompass both P01730 and CD8 epitopes of insulin . Disabling the mitotic spindle and tumor growth by targeting a cavity-induced allosteric site of survivin . Survivin is a member of the inhibitor of apoptosis protein family and has an essential role in mitosis . Survivin is overexpressed in a large variety of human cancers and represents an attractive target for cancer therapy . P00533 and Her/neu-transformed human tumors in particular exhibit high levels of survivin . The survivin protein forms dimers through a conserved region that is critical for subcellular localization and biological functions of the protein . We identified small molecules that target a specific cavity adjacent to the survivin dimerization surfaces . P28222 , a lead compound identified in the screen , can bind to the survivin protein at the intended target site . Moreover , P28222 alters spindle formation , causing mitotic arrest and cell death , and inhibits tumor growth in vitro and in vivo . Cell death occurs in premetaphase stage following mitotic arrest and is not a consequence of general toxicity . Thus , the study validates a novel therapeutic target site in the survivin protein and provides a promising strategy to develop a new class of therapeutic small molecules for the treatment of human cancers . Activation of critical , host-induced , metabolic and stress pathways marks neutrophil entry into cystic fibrosis lungs . Cystic fibrosis ( CF ) patients undergo progressive airway destruction caused in part by chronic neutrophilic inflammation . While opportunistic pathogens infecting CF airways can cause inflammation , we hypothesized that host-derived metabolic and stress signals would also play a role in this process . We show that neutrophils that have entered CF airways have increased phosphorylation of the eukaryotic initiation factor 4E and its partner the 4E-binding protein 1 ; 2 key effectors in the growth factor- and amino acid-regulated mammalian target of rapamycin ( P42345 ) pathway . Furthermore CF airway neutrophils display increased phosphorylation of the DB02527 response element binding protein ( CREB ) , a major transcriptional coactivator in stress signaling cascades . These active intracellular pathways are associated with increased surface expression of critical adaptor molecules , including the growth factor receptor CD114 and the receptor for advanced glycation end-products ( RAGE ) , a CREB inducer and sensor for host-derived damage-associated molecular patterns ( DAMPs ) . Most CF airway fluids lack any detectable soluble RAGE , an inhibitory decoy receptor for DAMPs . Concomitantly , CF airway fluids displayed high and consequently unopposed levels of P80511 ; a potent mucosa- and neutrophil-derived DAMP . CF airway neutrophils also show increased surface levels of 2 critical CREB targets , the purine-recycling enzyme P49961 and the multifunctional , P42345 -inducing P61073 receptor . This coordinated set of events occurs in all patients , even in the context of minimal airway inflammation and well-preserved lung function . Taken together , our data demonstrate an early and sustained activation of host-responsive metabolic and stress pathways upon neutrophil entry into CF airways , suggesting potential targets for therapeutic modulation . Urgosedin inhibits hypotension , hypoglycemia , and pro-inflammatory mediators induced by lipopolysaccharide . Urgosedin is a newly synthesized compound especially with serotonergic and alpha-adrenergic blocking actions . In rat isolated thoracic aorta , urgosedin competitively antagonized norepinephrine- , clonidine- , and serotonin-induced vasocontractions in a concentration-dependent manner . In radioligand binding experiments , urgosedin had significant binding affinities on alpha1/alpha2 , P08908 , P28222 and 5- Q13049 receptors . Intravenous injection of lipopolysaccharide ( LPS ) produced a biphasic hypotension in normotensive rats . Although intravenous injection of urgosedin caused minor depressor actions in the normotensive Wistar rat , urgosedin significantly attenuated the secondary prolonged hypotension produced by LPS . The plasma levels of cytokines ( IL-1beta , P05231 , P01375 , and P01579 ) and hypoglycemia induced by LPS were also reduced by urgosedin . Moreover , the acute survival rates ( 350 minutes ) of endotoxic shock increased from 0 % ( LPS group ) to 100 % in the groups pretreated with urgosedin . In RAW264.7 cells , urgosedin inhibited LPS-induced inducible nitric oxide synthase ( P35228 ) expression . In conclusion , our data suggest that urgosedin was a newly potent serotonergic and mild alpha-adrenergic blocking agent . Its prevention of LPS-induced hypotension and hypoglycemia might partially mediate through its inhibition activities on the P35228 expression and cytokines formation . Urgosedin might be an effective pharmacological agent against LPS-induced hypotension , hypoglycemia , and the formation of pro-inflammatory mediators . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Localization of P23219 and P35354 in the intracranial dura mater of the rat . Primary headaches such as migraine can be aborted by systemic administration of non-steroidal anti-inflammatory drugs ( NSAIDs ) , potentially through the non-selective inhibition of cyclooxygenase ( P36551 ) activity in the intracranial meninges . In this study we have used single and double labeling immunohistochemistry to examine the distribution of the P23219 and P35354 isoforms in the intracranial dura mater of the rat and identify cell types that express them . P23219 immunoreactivity was found in medium and small dural blood vessels and was co-expressed with the endothelial cell markers vimentin and the endothelial isoform of nitric oxide synthase ( ecNOS ) . P23219 was also found to be present in most dural mast cells . P35354 was mainly expressed in ED2-positive resident dural macrophages . Constitutive P35354 expression was also found in some axonal profiles , many of which were co-labeled with the nociceptor peptide marker P80511 . The findings suggest that NSAIDs may abort headache , at least in part , by inhibiting either neuronal or non-neuronal P36551 activity in the dura mater . Anandamide regulates neuropeptide release from capsaicin-sensitive primary sensory neurons by activating both the cannabinoid 1 receptor and the vanilloid receptor 1 in vitro . The effect of anandamide , which activates both the cannabinoid 1 ( P21554 ) receptor and the vanilloid receptor 1 ( Q8NER1 ) , was studied on calcitonin gene-related peptide ( P80511 ) release from cultured primary sensory neurons , the majority of which coexpress the P21554 receptor and Q8NER1 . Concentrations of anandamide < 1 micro m produced a small but significant P21554 receptor-mediated inhibition of basal P80511 release while higher concentrations induced Q8NER1 -mediated P80511 release . The excitatory effect of anandamide was potentiated by the P21554 receptor antagonist SR141716A . In the presence of SR141716A at concentrations < 100 nm , anandamide was equipotent with capsaicin in stimulating P80511 release . However , at higher concentrations anandamide produced more P80511 release than equimolar concentrations of capsaicin . Three and ten nanomolar anandamide inhibited the capsaicin-evoked P80511 release . In the presence of SR141716A , treatments which activated protein kinase A , protein kinase C and phospholipase C significantly potentiated the anandamide-evoked P80511 release at all anandamide concentrations . Although this potentiation was reduced when the P21554 receptor antagonist was omitted from the buffer , the P80511 release evoked by 300 nm and 1 micro m anandamide was still significantly larger than that seen with nonpotentiated cells . These data indicate that anandamide may regulate P80511 release from capsaicin-sensitive primary sensory neurons in vivo , and that the net effect of anandamide on transmitter release from capsaicin-sensitive primary sensory neurons depends on the concentration of anandamide and the state of the P21554 receptor and Q8NER1 . These findings also suggest that anandamide could be one of the molecules responsible for the development of inflammatory heat hyperalgesia . Serotonergic modulation of the acoustic startle response in rats during preweaning development . The involvement of serotonin ( 5-HT ) in modulating the acoustic startle response ( ASR ) is well established in adult rats , but 5-HT involvement during the preweaning period , when 5-HT neurons undergo extensive development , has not previously been described . Three 5-HT receptor subtypes are reported to modulate the ASR in adult rats : P08908 and 5-HT2 receptor agonists facilitate the ASR , whereas P28222 agonists decrease the response . In the present study , the effects of 5-HT agonists and generalized 5-HT depletion on the ASR were studied in preweanling animals , using independent groups of Long-Evans rats tested on postnatal day ( P01160 ) 13 , 17 and 21 . 8-Hydroxy-2-(di-n-propylamino) tetralin ( 8OHDPAT , 62-1000 micrograms/kg ) , a P08908 receptor agonist , and 5-methoxy-N,N-dimethyl tryptamine ( MeODMT , 2-4 mg/kg ) , a nonselective 5-HT agonist , had no effect on P01160 13 and then increased the ASR on P01160 17 and 21 . The 5-HT2 receptor antagonists cyproheptadine ( 5 mg/kg ) and ketanserin ( 5 mg/kg ) blocked the effect of MeODMT at both ages , providing some evidence that MeODMT increased the ASR through 5-HT2 receptors . 1-(m-Chlorophenyl) piperazine ( mCPP , 1-5 mg/kg ) , a P28222 agonist , had no effect on ASR amplitude on P01160 13 or 17 and then produced a dose-related decrease in the response on P01160 21 . Generalized depletion of 5-HT by 80-90 % in whole-brain and spinal cord , using p-chlorophenylalanine ( PCPA , 300 mg/kg 24 hr prior to testing ) , did not alter ASR amplitude at any age. ( ABSTRACT TRUNCATED AT 250 WORDS ) [ Pathophysiological basis of 3 subtypes in ganfeng neidong syndrome ] . The multiple parameters of 3 Subtypes : Ganyang Huafeng Syndrome ( GYHFS ) , Xuexu Shengfeng Syndrome and Yinxu Fengdong Syndrome of Ganfeng Neidong Syndrome were determined for the 1st time . It was found that there were several characteristics in GYHFS . ( 1 ) Disturbance of the cerebral blood flow and the damage of brain tissue was manifested by the abnormality of the bulbar conjunctival microcirculation , carotid Doppler ultrasonic determination and brainstem auditory and visual pathway , high blood viscosity , dysmnesia , free radical and lipid peroxidation injury and the changes of Zn , Cu , K and Mg after brain damage . ( 2 ) Stress status were expressed by the high plasma levels of cortisol , norepinephrine and epinephrine , decreased serum triiodothyronine level and hyperfunction of sympathetic nerve . ( 3 ) The marked changes of the regulating substance of the vessel smooth muscle function including the increased plasma levels of TXB2 , TXB2/6-k-PGF1 alpha , and calmodulin , as well as decreased SP , P01160 , P80511 . Other 2 subtypes had about the same changes of these parameters , but of milder disorders . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Dual P00533 and P42345 targeting in squamous cell carcinoma models , and development of early markers of efficacy . The epidermal growth factor receptor ( P00533 ) is a validated target in squamous cell carcinoma ( SCC ) of the head and neck . Most patients , however , do not respond or develop resistance to this agent . P42345 ( P42345 ) is involved in the pathogenesis of SCC of the head and neck ( SCCHN ) . This study aimed to determine if targeting P42345 in combination with P00533 is effective in SCC , and to develop early pharmacodynamic markers of efficacy . Two SCC cell lines , one resistant ( HEP2 ) and one of intermediate susceptibility ( Detroit 562 ) to P00533 inhibitors , were xenografted in vivo and treated with an P42345 inhibitor ( temsirolimus ) , an P00533 inhibitor ( erlotinib ) or a combination of both . DB06287 exerted superior growth arrest in both cell lines than erlotinib . The combined treatment resulted in synergistic antitumor effects in the Detroit 562 cell line . Immunohistochemical assessment of pharmacodynamic effects in fine-needle aspiration ( FNA ) biopsies early after treatment using phospho MAPK , Phospho-P70 and Ki67 as end points demonstrated pathway abrogation in the Detroit 562 tumours treated with the combination , the only group where regressions were seen . In conclusion , an P42345 inhibitor showed antitumor activity in P00533 -resistant SCC cell lines . Marked antitumor effects were associated with dual pathway inhibition , which were detected by early FNA biopsies . Efficacy , safety and tolerability of oral eletriptan in the acute treatment of migraine : results of a phase III , multicentre , placebo-controlled study across three attacks . The efficacy , safety and tolerability of the P28222 /D receptor agonist eletriptan ( 40 mg and 80 mg ) in acute treatment of migraine was evaluated in a multinational , randomized , double-blind , parallel-group , placebo-controlled , three-attack study treating 1153 patients . In the initial attack , significantly more eletriptan patients reported headache relief and complete pain relief at 2 h vs. placebo ( 40 mg 62 % and 32 % , 80 mg 65 % and 34 % , placebo 19 % and 3 % ; P < 0.0001 ) . Headache relief occurred faster after eletriptan , with more patients at both doses reporting relief 30 min ( P < 0.01 ) and 1 h ( P < 0.0001 ) after treatment than after placebo . There was a significantly lower recurrence rate with eletriptan 80 mg compared with placebo ( P < 0.01 ) . Adverse events for all treatments were generally mild or moderate and self-limiting . DB00216 40 mg and eletriptan 80 mg both appear to be effective and well-tolerated acute migraine treatments . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development .

MH_train_50

MH_train_50

MH_train_50

interacts_with DB06813?

multiple_choice

Genome-wide analysis of DNA copy number alterations and gene expression in gastric cancer . Genomic copy number aberrations ( CNAs ) are believed to play a major role in the development and progression of human cancers . Although many CNAs have been reported in gastric cancer , their genome-wide transcriptional consequences are poorly understood . In this study , to reveal the impact of CNAs on genome-wide expression in gastric cancer , we analysed 30 cases of gastric cancers for their CNAs by array comparative genomic hybridization ( array CGH ) and 24 of these 30 cases for their expression profiles by oligonucleotide-expression microarray . We found that with the application of laser microdissection , most CNAs were detected at higher frequency than in previous studies . Notably , gain at 20q13 was detected in almost all cases ( 97 % ) , suggesting that this may play an important role in the pathogenesis of gastric cancer . By comparing the array CGH data with expression profiles of the same samples , we showed that both genomic amplification and deletion strongly influence the expression of genes in altered genomic regions . Furthermore , we identified 125 candidate genes , consisting of 114 up-regulated genes located in recurrent regions ( > 10 % ) of amplification and 11 down-regulated genes located in recurrent regions of deletion . Up-regulation of several candidate genes , such as Q99741 , P60059 , Q9BTT0 , Q13895 and P37268 , was confirmed by immunohistochemistry . Interestingly , some candidate genes were localized at genomic loci adjacent to well-known genes such as P00533 , P04626 and Q13485 , and concordantly deregulated by genomic alterations . Based on these results , we propose that our list of candidate genes may contain novel genes involved in the pathogenesis of advanced gastric cancer . Single agent and combination studies of pralatrexate and molecular correlates of sensitivity . BACKGROUND : DB06813 is a dihydrofolate reductase ( P00374 ) inhibitor with high affinity for reduced folate carrier 1 ( P41440 -1 ) and folylpolyglutamate synthetase ( Q05932 ) , resulting in extensive internalization and accumulation in tumour cells . DB06813 is approved in the US for the treatment of relapsed or refractory peripheral T-cell lymphoma and is being investigated in various malignancies . Here , we evaluated molecular correlates of sensitivity to pralatrexate and explored combinations with a variety of anticancer agents . METHODS : Antiproliferative effects of pralatrexate were evaluated in 15 human-cancer cell lines using the MTT assay . Gene expression was evaluated using qRT-PCR . RESULTS : DB06813 and methotrexate had a similar pattern of cytotoxicity , pralatrexate being more potent . DB06813 potentiated the effects of platinum drugs , antimetabolites and P00533 inhibitors . Dose- and time-dependent cytotoxicity of pralatrexate correlated with high mRNA expression of Q05932 . Acquired resistance to pralatrexate was associated with decreased P41440 -1 expression , whereas methotrexate resistance correlated with increased P00374 expression , suggesting different mechanisms of acquired resistance . CONCLUSION : DB06813 was more potent than methotrexate in a panel of solid tumour lines . Our findings support the further clinical development of pralatrexate in combination with certain cytotoxics and targeted therapies , and suggest that P41440 -1 , Q05932 and P00374 may be potential biomarkers of outcome . Phosphodiesterase-4 influences the PKA phosphorylation status and membrane translocation of G-protein receptor kinase 2 ( P25098 ) in P29320 -293beta2 cells and cardiac myocytes . Membrane-recruitment of P25098 ( G-protein receptor kinase 2 ) provides a fundamental step in the desensitization process controlling GPCRs ( G-protein-coupled receptors ) , such as the beta2AR ( beta2-adrenergic receptor ) . In the present paper , we show that challenge of P29320 -293beta2 [ human embryonic kidney cells stably overexpressing the FLAG-tagged beta2AR-GFP ( green fluorescent protein ) ] cells with the beta-adrenoceptor agonist , isoprenaline , causes P25098 to become phosphorylated by PKA ( DB02527 -dependent protein kinase ) . This action is facilitated when DB02527 -specific DB05876 ( phosphodiesterase-4 ) activity is selectively inactivated , either chemically with rolipram or by siRNA ( small interfering RNA ) -mediated knockdown of Q07343 and Q08499 . DB05876 -selective inhibition by rolipram facilitates the isoprenaline-induced membrane translocation of P25098 , phosphorylation of the beta2AR by P25098 , membrane translocation of beta-arrestin and internalization of beta2ARs . DB05876 -selective inhibition also enhances the ability of isoprenaline to trigger the PKA phosphorylation of P25098 in cardiac myocytes . In the absence of isoprenaline , rolipram-induced inhibition of DB05876 activity in P29320 -293beta2 cells acts to stimulate PKA phosphorylation of P25098 , with consequential effects on P25098 membrane recruitment and P25098 -mediated phosphorylation of the beta2AR . We propose that a key role for DB05876 enzymes is : ( i ) to gate the action of PKA on P25098 , influencing the rate of P25098 phosphorylation of the beta2AR and consequential recruitment of beta-arrestin subsequent to beta-adrenoceptor agonist challenge , and ( ii ) to protect P25098 from inappropriate membrane recruitment in unstimulated cells through its phosphorylation by PKA in response to fluctuations in basal levels of DB02527 . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . DB06813 : a novel synthetic antifolate for relapsed or refractory peripheral T-cell lymphoma and other potential uses . PURPOSE : The pharmacology , pharmacokinetics , clinical trials , adverse effects , dosage , and economic considerations of pralatrexate ( DB06813 ) are reviewed . SUMMARY : Peripheral T-cell lymphoma ( PTCL ) comprises approximately 15-20 % of all aggressive lymphomas and 5-10 % of all non-Hodgkin 's lymphomas . Advanced PTCL is often refractory to traditional first-line chemotherapy regimens . DB06813 was developed as a synthetic folate analog antimetabolite that competitively inhibits dihydrofolate reductase ( P00374 ) . This results in the depletion of thymidine , leading to interference with deoxyribonucleic acid synthesis and cancer cell death . DB06813 has a higher potency than methotrexate and edatrexate ( EDX ) . The efficacy and safety of DB06813 have been demonstrated in the PROPEL trial , a prospective phase II trial in patients with relapsed or refractory PTCL . Patients with prior stem cell transplantation receiving DB06813 also had similar response rates ( RRs ) . DB06813 was investigated on the treatment of relapsed or refractory cutaneous T-cell lymphoma , previously treated advanced non-small cell lung cancer and other solid malignancies . DB06813 has similar side effects to other P00374 inhibitors . The most common side effect of DB06813 is mucositis . The recommended dose of DB06813 is 30 mg/m(2) weekly once for 6 weeks in 7-week cycle until disease progresses or unacceptable toxicity for PTCL and may require dose reduction or discontinuation . Patients should be supplemented with oral folic acid and intramuscular vitamin B(12) injections . CONCLUSION : DB06813 provides clinical benefit to patients with relapsed or refractory PTCL with durable complete and partial responses in patients who had not responded to multiple prior treatment regimens . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . C. elegans vulval development as a model system to study the cancer biology of P00533 signaling . Molecular genetic studies of C. elegans vulval development have helped to define an evolutionarily conserved signaling pathway from an P01133 -like ligand through P01133 -receptor , Ras and Q96HU1 kinase to the nucleus . Further studies have identified novel positive regulators such as Q8IVT5 -1 and Q09428 -8/ Q5T124 -2 and negative regulators such as cbl/SLI-1 . The many negative regulatory proteins might serve to prevent inappropriate signaling , and thus are analogous to tumor suppressor genes . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Second generation sequencing of the mesothelioma tumor genome . The current paradigm for elucidating the molecular etiology of cancers relies on the interrogation of small numbers of genes , which limits the scope of investigation . Emerging second-generation massively parallel DNA sequencing technologies have enabled more precise definition of the cancer genome on a global scale . We examined the genome of a human primary malignant pleural mesothelioma ( MPM ) tumor and matched normal tissue by using a combination of sequencing-by-synthesis and pyrosequencing methodologies to a 9.6X depth of coverage . Read density analysis uncovered significant aneuploidy and numerous rearrangements . Method-dependent informatics rules , which combined the results of different sequencing platforms , were developed to identify and validate candidate mutations of multiple types . Many more tumor-specific rearrangements than point mutations were uncovered at this depth of sequencing , resulting in novel , large-scale , inter- and intra-chromosomal deletions , inversions , and translocations . Nearly all candidate point mutations appeared to be previously unknown SNPs . Thirty tumor-specific fusions/translocations were independently validated with PCR and Sanger sequencing . Of these , 15 represented disrupted gene-encoding regions , including kinases , transcription factors , and growth factors . One large deletion in Q8N608 resulted in altered transcription and expression of Q8N608 transcripts in a set of 53 additional MPM tumors correlated with survival . Additionally , three point mutations were observed in the coding regions of Q9C056 , a transcription regulator , and Q6P4R8 , a DNA-binding protein involved in modulating P19838 . Several regions containing genes such as Q9H0N5 and P00374 , which are involved in growth factor signaling and nucleotide synthesis , respectively , were selectively amplified in the tumor . Second-generation sequencing uncovered all types of mutations in this MPM tumor , with DNA rearrangements representing the dominant type . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . Distinct mechanistic activity profile of pralatrexate in comparison to other antifolates in in vitro and in vivo models of human cancers . PURPOSE : This study evaluated mechanistic differences of pralatrexate , methotrexate , and pemetrexed . METHODS : Inhibition of dihydrofolate reductase ( P00374 ) was quantified using recombinant human P00374 . Cellular uptake and folylpolyglutamate synthetase ( Q05932 ) activity were determined using radiolabeled pralatrexate , methotrexate , and pemetrexed in NCI-H460 non-small cell lung cancer ( NSCLC ) cells . The tumor growth inhibition ( TGI ) was assessed using MV522 and NCI-H460 human NSCLC xenografts . RESULTS : Apparent K ( i ) values for P00374 inhibition were 45 , 26 , and > 200 nM for pralatrexate , methotrexate , and pemetrexed , respectively . A significantly greater percentage of radiolabeled pralatrexate entered the cells and was polyglutamylatated relative to methotrexate or pemetrexed . In vivo , pralatrexate showed superior anti-tumor activity in both NSCLC models , with more effective dose-dependent TGI in the more rapidly growing NCI-H460 xenografts . CONCLUSIONS : DB06813 demonstrated a distinct mechanistic and anti-tumor activity profile relative to methotrexate and pemetrexed . DB06813 exhibited enhanced cellular uptake and increased polyglutamylation , which correlated with increased TGI in NSCLC xenograft models . Effects of retroviral-mediated P08183 expression on hematopoietic stem cell self-renewal and differentiation in culture . Ex vivo expansion of hematopoietic stem cells would be useful for bone marrow transplantation and gene therapy applications . Toward this goal , we have investigated whether retrovirally-transduced murine stem cells could be expanded in culture with hematopoietic cytokines . Bone marrow cells were transduced with retroviral vectors expressing either the human multidrug resistance 1 gene ( HaMDR1 ) , a variant of human dihydrofolate reductase ( HaDHFR ) , or both P08183 and P00374 in an internal ribosomal entry site ( IRES ) -containing bicistronic vector ( HaMID ) . Cells were then expanded for 15 days in cultures stimulated with interleukin ( IL ) -3 , P05231 , and stem cell factor . When very low marrow volumes were injected into lethally irradiated recipient mice , long-term reconstitution with 100 % donor cells was seen in all mice injected with HaMDR1- or HaMID-transduced cells . By contrast , engraftment with HaDHFR- or mock-transduced cells ranged from partial to undetectable despite injection of significantly larger marrow volumes . In addition , mice transplanted with expanded HaMDR1- or HaMID-transduced stem cells developed a myeloproliferative disorder that was characterized by an increase in abnormal peripheral blood leukocytes . These results show that P08183 -transduced stem cells can be expanded in vitro with hematopoietic cytokines , but indicate that an increased stem cell division frequency can lead to stem cell damage . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . Evaluation of the pharmacokinetics , preclinical and clinical efficacy of pralatrexate for the treatment of T-cell lymphoma . INTRODUCTION : Peripheral T-cell lymphomas ( PTCLs ) are a heterogeneous group of T-cell neoplasms . Most patients with PTCL have a poor outcome with conventional therapies and are not cured without stem-cell transplantation . DB06813 , a novel antifolate chemotherapeutic agent , was rationally designed to impede folate metabolism by inhibiting dihydrofolate reductase ( P00374 ) and to be more efficiently internalized into tumor cells . DB06813 is the first drug that is FDA approved for patients with relapsed and refractory PTCL . AREAS COVERED : DB06813 has been used as a single agent and in combination with other agents in clinical trials for non-Hodgkin 's lymphoma and Hodgkin 's disease as well as in solid tumors . This review will cover the development of pralatrexate , the pharmacokinetics of pralatrexate , preclinical findings with pralatrexate and clinical studies of pralatrexate in hematologic malignancies . EXPERT OPINION : DB06813 has significant activity in vitro , and in early Phase I/II trials , responses were noted in patients with aggressive T-cell lymphomas . The DB06813 in Patients with Relapsed or Refractory Peripheral T-Cell Lymphoma trial demonstrated the activity of pralatrexate across a spectrum of heavily pretreated patients with different aggressive T-cell lymphoma subtypes , and studies in cutaneous T-cell lymphoma have shown efficacy at different doses and schedules . The most frequent adverse events in these trials were mucositis , reversible thrombocytopenia and fatigue . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . DB00227 overcomes gefitinib resistance in human non-small cell lung cancer cells with K-Ras mutations . DB00227 is a 3-hydroxy-3-methylglutaryl-coenzyme A ( HMG- DB01992 ) reductase inhibitor . Its inhibitory action on P04035 leads to depletion of isoprenoids , which inhibits post-translational modification of DB01367 . In this study , we investigated the effect of combining lovastatin with gefitinib on gefitinib-resistant human non-small cell lung cancer ( NSCLC ) cell lines with K-Ras mutations . Antitumor effects were measured by growth inhibition and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay . Effects on apoptosis were determined by flow cytometry , DNA fragmentation , and immunoblots . Protein levels of DB01367 , AKT/pAKT , and RAF/ P27361 /2 in cancer cells were analyzed by immunoblot . Compared with gefitinib alone , a combination of gefitinib with lovastatin showed significantly enhanced cell growth inhibition and cytotoxicity in gefitinib-resistant A549 and NCI-H460 human NSCLC cells . In addition , lovastatin combination treatment significantly increased gefitinib-related apoptosis , as determined by fluorescence microscopy and flow cytometric analysis . These effects correlated with up-regulation of cleaved caspase-3 , poly ( ADP-ribose ) polymerase ( PARP ) , and Bax and down-regulation of Bcl-2 . The combination of lovastatin and gefitinib effectively down-regulated DB01367 protein and suppressed the phosphorylation of RAF , P27361 /2 , AKT , and P00533 in both cell lines . Taken together , these results suggest lovastatin can overcome gefitinib resistance , in NSCLC cells with K-Ras mutations , by down regulation of DB01367 protein , which leads to inhibition of both RAF/ P29323 and AKT pathways . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Suppression of tumor growth and metastasis by a P17948 antagonizing peptide identified from a phage display library . Although the P15692 -Flk-1-pathway has been known as the major driving force of angiogenesis , new evidence has shown that P17948 /Flt-1 plays important roles during the neovascularization under pathological conditions including tumor , atherosclerosis and arthritis . In search of Flt-1 receptor antagonizing peptides , we screened a phage display 12-mer-peptide library with recombinant Flt-1 protein . Seven candidate peptides were identified that specifically bound to P15692 receptor Flt-1 , of which peptide F56 ( WHSDMEWWYLLG ) almost abolished P15692 binding to receptor Flt-1 in vitro . In vivo , F56 fused with P00374 ( P00374 -F56 ) inhibited angiogenesis in a P62158 assay . Moreover , P00374 -F56 significantly inhibited the growth of nodules of human gastric cancer cell line MGC-803 in BALB/c nude mice . Histological analyses showed that necrosis of the implanted tumor was markedly enhanced following treatment with P00374 -F56 . In the severe combined immunodeficiency disease ( SCID ) mouse model for studying metastasis of the human breast cancer cell line BICR-H1 , synthetic peptide F56 significantly inhibited tumor growth and lung metastases . Taken together , our results have demonstrated that peptide F56 , as a Flt-1 receptor antagonist , fulfilled the antiangiogenic and antimetastatic effects by specifically interfering with the interaction between P15692 and receptor Flt-1 . Thus , short peptide F56 may have clinical potential in tumor therapy . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Analyses of cross species polymerase chain reaction products to infer the ancestral state of human polymorphisms . In numerous population genetic and disease association studies decisions about the ancestry of polymorphic alleles are often made based on the relative frequency of the alleles in the extant populations with the most frequent allele being deemed as ancestral . However , the frequency of an allele in a population is generally not a perfect indicator of its ancestral status . A more accurate method to assess ancestral/derived status of polymorphic alleles involves identification of shared alleles between species . We used this strategy to examine genomic regions homologous to several human polymorphisms in four species of non-human primates . Cross species polymerase chain reaction ( CS-PCR ) , with primers designed from human sequence , was used to investigate regions of interest . Nineteen polymorphisms at six loci ( P14416 , HOXB@ , PAH , D4S10 , P10745 , and P07949 ) were examined either by restriction fragment length analysis of PCR products ( PCR-RFLP ) or by direct sequencing . At seventeen of the eighteen PCR-RFLPs , non-human primates were monomorphic and identical to each other for either lack of restriction enzyme site or presence of the site . Thus , at these seventeen polymorphic sites the shared alleles are most likely to be the ancestral ones in humans . In several cases we have used sequence data to further demonstrate that the nucleotide at the site of the polymorphism is conserved between species confirming the hypothesis of a single ancestral allele . However , not all human alleles can be simply resolved into ancestral and derived ; sequence data from one PCR-RFLP ( in an intron of the PAH locus ) and a single strand conformational polymorphism ( SSCP ) in the 3' untranslated region ( UTR ) of the P14416 gene illustrate this point . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . In vitro and in vivo biological activities of a novel nonpolyglutamable anti-folate , MX-68 . MX-68 is a newly synthesized anti-folate , chemically designed not to undergo intracellular polyglutamation and to have increased affinity to dihydrofolate reductase ( P00374 ) . In the present study , we examined the in vitro and in vivo biological activities of MX-68 compared with methotrexate ( MTX ) which forms several polyglutamates intracellularly . MX-68 dose-dependently inhibited the proliferation of PHA- , anti-CD3- , or PMA plus ionomycin-stimulated peripheral blood mononuclear cells ( PBMC ) and endothelial cells ( EC ) from normal subjects as well as P01584 - or P01375 alpha-stimulated synovial fibroblastic cells ( SC ) from rheumatoid arthritis ( RA ) patients . Coaddition of folinic acid completely reversed the anti-proliferative effects of both MX-68 and MTX . Although the anti-proliferative activities of MX-68 were almost comparable to those of MTX , the washout study clearly showed the characteristic nature of MX-68 . When drugs were removed during culture , the suppressive effect of MX-68 completely disappeared , whereas suppression by MTX was merely weakened . MX-68 dramatically suppressed the onset of collagen-induced arthritis ( CIA ) in mice when the drug was orally administered three times a week. starting from the day of first immunization . In this model , 2 mg/kg of MX-68 was sufficient to completely suppress arthritis , whereas suppression by the same dose of MTX was partial . These lines of evidence suggest that polyglutamation is not always a prerequisite in the anti-rheumatic effects of anti-folate . In addition , since intracellular accumulation of polyglutamates is thought to have adverse effects , MX-68 may become a more potent and less toxic anti-rheumatic drug than MTX . Development of stable cell lines expressing different subtypes of GABAA receptors . The experiments reported here were motivated by our interest to express in stably-transfected cells large amounts of recombinant rat GABAA receptors . For this , we developed an original two step selection strategy , in which the first step consisted of transfecting P29320 293 cells with rat GABAA receptor alpha and beta subunits . G 418 resistant colonies isolated at this step were screened for [ 3H ] muscimol binding to select for those that coexpressed alpha- and beta-subunits . The best alpha and beta subunit expressing colony was then supertransfected with a plasmid coding for the gamma rat GABAA receptor subunit and a mutant P00374 gene . After a second round of selection , this time in presence of methotrexate , those colonies that coexpressed ternary alpha beta gamma GABAA receptor combinations were distinguished using [ 3H ] flumazenil as a probe . This strategy was applied to the isolation of 3 GABAA receptor clones , alpha 1 beta 2 gamma 2s , alpha 3 beta 2 gamma 2s and alpha 5 beta 3 gamma 2s , that expressed relatively high levels of these proteins . These 3 cell lines exhibited pharmacological and functional properties similar to cells transiently-transfected with equivalent subunit combinations . These cell lines therefore provide attractive models with which to evaluate the intrinsic activity and potency of compounds at recombinant GABAA receptor subtypes . [ Presynaptic P08908 receptors in immunomodulation ] . It is shown that a selective agonist of P08908 receptors 8-OH-DPAT in a low dose ( 0.1 mg/kg ) , which is known to affect mainly the presynaptic P08908 receptors increased the immune response at the peak of reactions ( the forth or fifth day after immunization with sheep red blood cells - SRBC ) in CBA mice and Wistar rats . Treatment of the animals with the drug 15 min prior to antigen injection raised the number of plaque-forming cells ( lgM- P27918 ) and rosette-forming cells ( P41440 ) in the spleen . The preliminary blockade of P08908 receptor with a selective antagonist of P08908 receptors WAY-100635 ( 0.1 mg/kg ) prevented the immunostimulating effect of 5-HT 1A receptors agonist 8-OH-DPAT , whereas WAY-100635 administration alone in the same dose did n't change the immune response . Activation of P08908 receptors under conditions of electrical lesion of 5-HTergic neurons of the nucleus raphe was unable to enhance the immune reactions , as it did in sham-operated rats . The data obtained indicate that the somatodendric P08908 autoreceptors are involved in immunomodulation . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Extracellular signal-regulated kinase/mitogen-activated protein kinases block internalization of delta-opioid receptors . Translocation of G protein-coupled receptors ( GPCRs ) from the cell membrane to cytosol depends on the kind of ligand activating the receptor . This principle is clearly demonstrated for opioid receptors , because diverse opiate agonists rapidly induce receptor internalization , whereas morphine almost fails . We report here the impact of mitogen-activated protein ( Q96HU1 ) kinase isoforms extracellular signal-regulated kinase ( P29323 )1/2 on the internalization of delta-opioid receptors ( DORs ) expressed in human embryonic kidney ( P29320 )293 cells . Receptor activation by etorphine turned out to transiently phosphorylate P29323 / Q96HU1 kinases and bring about Q8IXH6 internalization within 20 min . In contrast , prolonged exposure of HEK293 cells to morphine excited persistent phosphorylation of P29323 / Q96HU1 kinases , and those cells failed to internalize the opioid receptor . When P29323 / Q96HU1 kinase phosphorylation was blocked by 2'-Amino-3'-methoxyflavone ( PD98059 ) , morphine gained the ability to strongly induce Q8IXH6 endocytosis . The importance of activated Q96HU1 kinases for Q8IXH6 internalization is further demonstrated by glutamate and paclitaxel because these substances induce phosphorylation of P27361 /2 and concomitantly prevent Q8IXH6 sequestration by etorphine . In addition , receptor internalization by morphine was facilitated by inhibition of protein kinase C and opioid-mediated transactivation of epidermal growth factor receptor ( P00533 ) , both activating P29323 / Q96HU1 kinases by opioids . The mechanism affording Q8IXH6 internalization by PD98059 may relate to arrestin , which uncouples GPCRs and thus triggers receptor internalization . Arrestin considerably translocates toward the cell membrane upon Q8IXH6 activation by morphine in presence of the Q96HU1 kinase blocker , but it fails in the absence of PD98059 . We conclude that P29323 / Q96HU1 kinase activity prevents opioid receptor desensitization and sequestration by blocking arrestin 2 interaction with activated DORs . A novel bisphosphonate inhibitor of squalene synthase combined with a statin or a nitrogenous bisphosphonate in vitro . Statins and nitrogenous bisphosphonates ( NBP ) inhibit 3-hydroxy-3-methylglutaryl-coenzyme-A reductase ( P04035 ) and farnesyl diphosphate synthase ( P14324 ) , respectively , leading to depletion of farnesyl diphosphate ( FPP ) and disruption of protein prenylation . P37268 ( P37268 ) utilizes FPP in the first committed step from the mevalonate pathway toward cholesterol biosynthesis . Herein , we have identified novel bisphosphonates as potent and specific inhibitors of P37268 , including the tetrasodium salt of 9-biphenyl-4,8-dimethyl-nona-3,7-dienyl-1,1-bisphosphonic acid ( compound 5 ) . Compound 5 reduced cholesterol biosynthesis and lead to a substantial intracellular accumulation of FPP without reducing cell viability in HepG2 cells . At high concentrations , lovastatin and zoledronate impaired protein prenylation and decreased cell viability , which limits their potential use for cholesterol depletion . When combined with lovastatin , compound 5 prevented lovastatin-induced FPP depletion and impairment of protein farnesylation . Compound 5 in combination with the NBP zoledronate completely prevented zoledronate-induced impairment of both protein farnesylation and geranylgeranylation . Cotreatment of cells with compound 5 and either lovastatin or zoledronate was able to significantly prevent the reduction of cell viability caused by lovastatin or zoledronate alone . The combination of an P37268 inhibitor with an P04035 or P14324 inhibitor provides a rational approach for reducing cholesterol synthesis while preventing nonsterol isoprenoid depletion . Signatures of positive selection in genes associated with human skin pigmentation as revealed from analyses of single nucleotide polymorphisms . Phenotypic variation between human populations in skin pigmentation correlates with latitude at the continental level . A large number of hypotheses involving genetic adaptation have been proposed to explain human variation in skin colour , but only limited genetic evidence for positive selection has been presented . To shed light on the evolutionary genetic history of human variation in skin colour we inspected 118 genes associated with skin pigmentation in the Perlegen dataset , studying single nucleotide polymorphisms ( SNPs ) , and analyzed 55 genes in detail . We identified eight genes that are associated with the melanin pathway ( Q9UMX9 , Q04671 , P17643 , P40126 , P21583 , P00533 , P14416 and Q03181 ) and presented significant differences in genetic variation between Europeans , Africans and Asians . In six of these genes we detected , by means of the EHH test , variability patterns that are compatible with the hypothesis of local positive selection in Europeans ( Q04671 , P17643 and P21583 ) and in Asians ( Q04671 , P40126 , P21583 , P00533 and P14416 ) , whereas signals were scarce in Africans ( P40126 , P00533 and P14416 ) . Furthermore , a statistically significant correlation between genotypic variation in four pigmentation candidate genes and phenotypic variation of skin colour in 51 worldwide human populations was revealed . Overall , our data also suggest that light skin colour is the derived state and is of independent origin in Europeans and Asians , whereas dark skin color seems of unique origin , reflecting the ancestral state in humans . A phase II study of pralatrexate with vitamin B12 and folic acid supplementation for previously treated recurrent and/or metastatic head and neck squamous cell cancer . BACKGROUND : DB06813 ( Fotolyn(TM) ; Allos Therapeutics Inc. ) is an antifolate dihydrofolate reductase ( P00374 ) inhibitor . We conducted a phase II study of pralatrexate with folic acid and B12 supplementation in patients with recurrent and/or metastatic head and neck squamous cell cancer ( R/M HNSCC ) . PATIENTS AND METHODS : This was a single-arm , Simon optimal two stage phase II study . Patients with R/M HNSCC previously treated with chemotherapy were eligible . The study was initiated with a dosing schedule of pralatrexate 190 mg/m(2) biweekly on a 4-week cycle with vitamin supplementation . Due to toxicity concerns , the dosing was modified to 30 mg/m(2) weekly for 3 weeks in a 4-week cycle with vitamin supplementation . Radiologic imaging was to be obtained about every 2 cycles . RESULTS : Thirteen subjects were enrolled ; 12 were treated . Seven of the twelve patients had previously received ≥2 lines of chemotherapy . The most common grade 3 toxicity was mucositis ( 3 patients ) . Seven patients did not complete two cycles of therapy due to progression of disease ( 4 ) , toxicity ( 1 ) , death ( 1 ) , and withdrawal of consent ( 1 ) . Two deaths occurred : one due to disease progression and the other was an unwitnessed event that was possibly related to pralatrexate . No clinical activity was observed . The median overall survival was 3.1 months . The study was closed early due to lack of efficacy . CONCLUSIONS : DB06813 does not possess clinical activity against previously treated R/M HNSCC . Evaluation of pralatrexate in other clinical settings of HNSCC management with special considerations for drug toxicity may be warranted . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Cigarette smoke synergistically enhances respiratory mucin induction by proinflammatory stimuli . Pathogenic factors associated with chronic obstructive pulmonary disease ( P48444 ) , such as cigarette smoke , proinflammatory cytokines , and bacterial infections , can individually induce respiratory mucins in vitro and in vivo . Since co-presence of these factors is common in lungs of patients with P48444 , we hypothesized that cigarette smoke can amplify mucin induction by bacterial exoproducts and proinflammatory cytokines , resulting in mucin hyperproduction . We demonstrated that cigarette smoke extract ( CSE ) synergistically increased gene expression and protein production of P98088 mucin induced by LPS or P01375 in human airway epithelial NCI-H292 cells . CSE also enhanced expression and production of P98088 mucin induced by epidermal growth factor receptor ( P00533 ) ligands TGF-alpha and amphiregulin , as well as LPS- and P01375 - induced expression and/or release of TGF-alpha and amphiregulin . Furthermore , ( 4-[(3-bromophenyl)amino]-6,7-diaminoquinazoline ) , a potent inhibitor of P00533 , blocked synergistic induction of P98088 mucin . H(2)O(2) mimicked the synergistic effects of CSE , while antioxidant N-acetyl-L-cysteine prevented synergistic induction of P98088 mucin by CSE . In a rat model of LPS-induced airway inflammation , concurrent cigarette smoke inhalation enhanced mucin content of the bronchoalveolar lavage fluid , muc5AC gene expression , and mucous cell metaplasia in the airways . These results suggest that cigarette smoke has the potential to synergistically amplify induction of respiratory mucins by proinflammatory stimuli relevant to P48444 pathogenesis and contribute to mucin hyperproduction observed in patients with P48444 .

MH_train_51

MH_train_51

MH_train_51

interacts_with DB00773?

multiple_choice

Inhibition of cholinergic signaling causes apoptosis in human bronchioalveolar carcinoma . Recent case-controlled clinical studies show that bronchioalveolar carcinomas ( BAC ) are correlated with smoking . DB00184 , the addictive component of cigarettes , accelerates cell proliferation through nicotinic acetylcholine receptors ( nAChR ) . In this study , we show that human BACs produce acetylcholine ( ACh ) and contain several cholinergic factors including acetylcholinesterase ( P22303 ) , choline acetyltransferase ( P28329 ) , choline transporter 1 ( Q9GZV3 , Q9GZV3 ) , vesicular acetylcholine transporter ( Q16572 , Q16572 ) , and nACh receptors ( AChRs , CHRNAs ) . DB00184 increased the production of ACh in human BACs , and ACh acts as a growth factor for these cells . DB00184 -induced ACh production was mediated by α7- , α3β2- , and β3-nAChRs , P28329 and Q16572 pathways . We observed that nicotine upregulated P28329 and Q16572 . Therefore , we conjectured that Q16572 antagonists , such as vesamicol , may suppress the growth of human BACs . Vesamicol induced potent apoptosis of human BACs in cell culture and nude mice models . Vesamicol did not have any effect on P01133 or insulin-like growth factor-II-induced growth of human BACs. siRNA-mediated attenuation of Q16572 reversed the apoptotic activity of vesamicol . We also observed that vesamicol inhibited Akt phosphorylation during cell death and that overexpression of constitutively active Akt reversed the apoptotic activity of vesamicol . Taken together , our results suggested that disruption of nicotine-induced cholinergic signaling by agents such as vesamicol may have applications in BAC therapy . Rad10 exhibits lesion-dependent genetic requirements for recruitment to DNA double-strand breaks in Saccharomyces cerevisiae . In the yeast Saccharomyces cerevisiae , the Rad1-Rad10 protein complex participates in nucleotide excision repair ( P55055 ) and homologous recombination ( HR ) . During HR , the Rad1-Rad10 endonuclease cleaves 3' branches of DNA and aberrant 3' DNA ends that are refractory to other 3' processing enzymes . Here we show that yeast strains expressing fluorescently labeled Rad10 protein ( Rad10-YFP ) form foci in response to double-strand breaks ( DSBs ) induced by a site-specific restriction enzyme , I-SceI or by ionizing radiation ( IR ) . Additionally , for endonuclease-induced DSBs , Rad10-YFP localization to DSB sites depends on both Q06609 and P43351 , but not P49959 while IR-induced breaks do not require Q06609 . Finally , Rad10-YFP colocalizes with Rad51- P27918 and with Rad52- P27918 at DSB sites , indicating a temporal overlap of Rad52 , Rad51 and Rad10 functions at DSBs . These observations are consistent with a putative role of Rad10 protein in excising overhanging DNA ends after homology searching and refine the potential role(s) of the Rad1-Rad10 complex in DSB repair in yeast . Novel Hsp90 inhibitor DB00238 -AUY922 radiosensitizes prostate cancer cells . Outcomes for poor-risk localized prostate cancers treated with radiation are still insufficient . Targeting the " non-oncogene " addiction or stress response machinery is an appealing strategy for cancer therapeutics . Heat-shock-protein-90 ( Hsp90 ) , an integral member of this machinery , is a molecular chaperone required for energy-driven stabilization and selective degradation of misfolded " client " proteins , that is commonly overexpressed in tumor cells . Hsp90 client proteins include critical components of pathways implicated in prostate cancer cell survival and radioresistance , such as androgen receptor signaling and the PI3K-Akt- P42345 pathway . We examined the effects of a novel non-geldanamycin Hsp90 inhibitor , AUY922 , combined with radiation ( RT ) on two prostate cancer cell lines , Myc-CaP and PC3 , using in vitro assays for clonogenic survival , apoptosis , cell cycle distribution , γ- P16104 foci kinetics and client protein expression in pathways important for prostate cancer survival and radioresistance . We then evaluated tumor growth delay and effects of the combined treatment ( RT-AUY922 ) on the PI3K-Akt- P42345 and AR pathways in a hind-flank tumor graft model . We observed that AUY922 caused supra-additive radiosensitization in both cell lines at low nanomolar doses with enhancement ratios between 1.4-1.7 ( p < 0.01 ) . RT-AUY922 increased apoptotic cell death compared with either therapy alone , induced G 2-M arrest and produced marked changes in client protein expression . These results were confirmed in vivo , where RT-AUY922 combination therapy produced supra-additive tumor growth delay compared with either therapy by itself in Myc-CaP and PC3 tumor grafts ( both p < 0.0001 ) . Our data suggest that combined RT-AUY922 therapy exhibits promising activity against prostate cancer cells , which should be investigated in clinical studies . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . Anti-angiogenic effects of the water extract of HangAmDan ( WEHAD ) , a Korean traditional medicine . We investigated the anti-angiogenic effects of the water extract of HangAmDan ( WEHAD ) , which is a crude extract of nine Korean medicinal substances of animal and plant origin . In human umbilical vein endothelial cells , WEHAD significantly inhibited P09038 -induced proliferation , adhesion , migration , and capillary tube formation . We used an antibody array to perform an analysis of signaling proteins , which showed up-regulated expression of various proteins including Q06609 , P43351 , and p73 , and down-regulated expression of pFAK . Blood vessel formation in a chick chorioallantoic membrane ( P62158 ) treated with WEHAD was markedly reduced in length compared with a PBS-treated control group . These results suggest that inhibition of angiogenesis by WEHAD may be the mechanism of action for the anti-cancer effects of HAD . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . Topoisomerase II-mediated DNA cleavage and mutagenesis activated by nitric oxide underlie the inflammation-associated tumorigenesis . AIMS : Both cancer-suppressing and cancer-promoting properties of reactive nitrogen and oxygen species ( RNOS ) have been suggested to play a role in tumor pathology , particularly those activities associated with chronic inflammation . Here , we address the impact of nitric oxide ( NO ) on the induction of DNA damage and genome instability with a specific focus on the involvement of topoisomerase II ( P11388 ) . We also investigate the contribution of NO to the formation of skin melanoma in mice . RESULTS : Similar to the P11388 -targeting drug , etoposide ( DB00773 ) , the NO-donor , S-nitrosoglutathione ( GSNO ) , induces skin melanomas formation in 7,12-dimethyl- benz[a]anthracene ( DMBA ) -initiated mice . To explore the mechanism(s) underlying this NO-induced tumorigenesis , we use a co-culture model system to demonstrate that inflamed macrophages with inducible NO synthase ( P35228 ) expression cause γ- P16104 activation , p53 phosphorylation , and chromosome DNA breaks in the target cells . Inhibitor experiments revealed that NO and P11388 isozymes are responsible for the above described cellular phenotypes . Notably , NO , unlike DB00773 , preferentially induces the formation of TOP2β cleavable complexes ( TOP2βcc ) in cells . Moreover , GSNO induced P11388 -dependent DNA sequence rearrangements and cytotoxicity . Furthermore , the incidences of GSNO- and DB00773 -induced skin melanomas were also observed to be lower in the skin-specific top2β-knockout mice . Our results suggest that P11388 isozymes contribute to NO-induced mutagenesis and subsequent cancer development during chronic inflammation . INNOVATION AND CONCLUSIONS : We provide the first experimental evidence for the functional role of P11388 in NO-caused DNA damage , mutagenesis , and carcinogenesis . Notably , these studies contribute to our molecular understanding of the cancer-promoting actions of RNOS during chronic inflammation . Substance P promotes expansion of human mesenteric preadipocytes through proliferative and antiapoptotic pathways . White adipose tissue is intimately involved in the regulation of immunity and inflammation . We reported that human mesenteric preadipocytes express the DB05875 ( SP ) -mediated neurokinin-1 receptor ( P25103 ) , which signals proinflammatory responses . Here we tested the hypothesis that SP promotes proliferation and survival of human mesenteric preadipocytes and investigated responsible mechanism(s) . Preadipocytes were isolated from mesenteric fat biopsies during gastric bypass surgery . Proliferative and antiapoptotic responses were delineated in 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium ( MTS ) , bromodeoxyuridine ( BrdU ) , caspase-3 , and TUNEL assays , as well as Western immunoanalysis . SP ( 10(-7) M ) increased MTS and proliferation ( BrdU ) and time dependently ( 15-30 min ) induced Akt , P01133 receptor , IGF receptor , integrin alphaVbeta3 , phosphatidylinositol 3-kinase , and PKC-theta phosphorylation . Furthermore , pharmacological antagonism of Akt and PKC-theta activation significantly attenuated SP-induced preadipocyte proliferation . Exposure of preadipocytes to the proapoptotic P48023 ( P48023 , 100 microM ) resulted in nuclear DNA fragmentation ( TUNEL assay ) , as well as increased cleaved poly ( ADP-ribose ) polymerase , cleaved caspase-7 , and caspase-3 expression . Cotreatment with SP almost completely abolished these responses in a P25103 -dependent fashion . SP ( 10(-7) M ) also time dependently stimulated expression 4E binding protein 1 and phosphorylation of P08133 S6 kinase , which increased protein translation efficiency . SP increases preadipocyte viability , reduces apoptosis , and stimulates proliferation , possibly via cell cycle upregulation and increased protein translation efficiency . SP-induced proliferative and antiapoptotic pathways in fat depots may contribute to development of the creeping fat and inflammation characteristic of Crohn 's disease . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Analysis of DNA recombination and repair proteins in living cells by photobleaching microscopy . DNA double strand break repair through homologous recombination has been shown biochemically to require the coordinated action of the P43351 group of proteins , including the DNA strand exchange protein Rad51 . We have started to develop experimental tools to investigate the close cooperation of homologous recombination proteins in living cells , where proteins operate in the context of chromatin and in the presence of other nuclear processes . This chapter describes in detail methods to establish cell lines stably expressing green fluorescent protein -tagged recombination proteins and photobleaching techniques to investigate the behavior of the proteins with the use of live cell video microscopy . Fluorescence recovery after photobleaching ( P42345 ) , fluorescence loss after photobleaching ( FLIP ) , and their combination in the same cell are useful techniques to gain insights into the dynamic behavior of the recombination proteins . Parameters such as diffusion rates and mobile versus immobile fractions before and after DNA damage induction can be obtained . In addition , residence times of recombination proteins at sites of DNA damage can be determined . Through the application of P42345 and FLIP it is possible to establish whether proteins are present in the same multiprotein complex , whether this is affected by DNA damage induction , and whether proteins dynamically associate with and dissociate from sites of DNA damage . mRNA expression , functional profiling and multivariate classification of colon biopsy specimen by cDNA overall glass microarray . AIM : To understand the local pathophysiological alterations and gene ontology-based functional classification of colonic biopsies into inflammatory and neoplastic diseases . METHODS : Total RNA was extracted from frozen biopsies and amplified by T7-method . Expression profile was evaluated by Atlas Glass 1K microarrays . After microarray quality control , applicable data were available from 10 adenomas , 6 colorectal adenocarcinomas ( CRCs ) , and 6 inflammatory bowel diseases ( IBDs ) . Multivariate statistical and cell functional analyses were performed . Real-time RT-PCR and immunohistochemistry were used for validation . RESULTS : Discriminant analysis of selected genes , could correctly reclassify all 22 samples using 4 parameters ( heat shock transcription factor-1 , bystin-like , calgranulin-A , O14798 ) . Q9UKU7 samples were characterized by overregulated chemokine ( C-X-C motif ) ligand 13 , replication protein A1 , Q15723 and downregulated Q9Y4K3 , P10415 -interacting killer genes . In adenomas upregulation of Q9Y4K3 , replication protein A1 , Q15723 and underexpression of P10415 -associated X protein , calgranulin-A genes were found . CRC cases had significantly increased epidermal growth factor receptor , topoisomerase-1 , v-jun , Q9Y4K3 and O14798 , and decreased Q06609 and P43351 DNA repair gene , protein phosphatase-2A and P10415 -interacting killer mRNA levels . P00533 RT-PCR and immunohistochemistry , topoisomerase-1 RT-PCR confirmed the chip results . CONCLUSION : Different histological alterations can be reclassified by functional , multivariate analysis using cDNA microarrays . Further studies with expanded sample number are needed for subclassification of pathological alterations . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Senescence-associated secretory phenotype in a mouse model of bleomycin-induced lung injury . DB00290 produces DNA damage , apoptosis and senescence , all of which play crucial roles in the development of pulmonary fibrosis . Recently , close attention has been paid to a DNA damage-induced phenotypic change ( senescence-associated secretory phenotype ; SASP ) as a trigger for the secretion of various mediators which modify the processes of tissue injury , inflammation , repair and fibrosis . We characterized the SASP in a murine model of bleomycin-induced lung injury . Mice were intratracheally administered bleomycin or control saline , and the lungs were obtained on days 7 , 14 and 21 . The occurrence of DNA damage and the SASP in the lungs was examined by immunostaining . γ P16104 immunostaining of the bleomycin-treated lungs revealed double-strand breaks ( DSBs ) , largely within P12830 -positive , β4-integirn-positive alveolar epithelial cells . The DSBs were associated with phosphorylation of Q13315 /ATR , a central signal transducer mediating the DNA damage response , and upregulation of the cyclin-dependent kinase inhibitor P38936 (CIP1) . The DSBs persisted for at least 21 days after the bleomycin exposure , although it began to wane after 7 days . A subpopulation of the γ P16104 -positive , DNA-damaged cells exhibited the SASP , characterized by overexpression of P05231 , TNFα , P08253 and P14780 , in association with the phosphorylation of IKKα/β and p38 MAPK . Persistent DNA damage and the SASP are induced in the process of bleomycin-induced lung injury and repair , suggesting that these events play an important role in the regulation of inflammation and tissue remodeling in bleomycin-induced pneumopathy . Expression and prognostic significance of apoptotic genes in oral squamous cell carcinoma . Oral squamous cell carcinoma ( OSCC ) is the most common malignancy of oral cavity . Human cancers are characterized by an imbalance of regulatory mechanisms controlling different cellular pathways , including apoptosis . Apoptosis occurs in a wide variety of physiological processes , such as embryonic development , tissue homeostasis or immune defense , and its role is to remove harmful , damaged , or unwanted cells . Defective apoptosis represents an important causative factor in the development/progression of cancer , and the ability of tumor cells to evade apoptosis can play a significant role in their resistance to conventional anticancer treatment . We investigated the expression profile of genes involved in the apoptotic mechanism in 21 paired tissue samples ( OSCC and adjacent normal oral mucosa ) by cDNA macroarray , in order to identify differentially expressed genes in oral cancer compared to normal tissue . To validate the results obtained by cDNA macroarray , quantitative real-time PCR , Western blot , and immunohistochemical analyses were performed . Results obtained by cDNA macroarray analysis showed different expression levels of P78560 , Q13158 , Q13315 , O14727 , and Q9H3D4 genes in OSCC compared to normal mucosa . Differential gene expression measurements ( tumor vs. normal tissue ) performed by real-time PCR showed an overexpression of Q13158 and a downregulation of Q13315 . Moreover , Western blot analysis confirmed that both P78560 and O14727 were decreased in OSCC compared to normal oral mucosa . As showed by immunohistochemistry , OSCC exhibited increased expression of p63 compared to normal tissue . Interestingly , a statistically significant positive correlation was found between p63 expression and the histological grade . PIKKs -- the solenoid nest where partners and kinases meet . The recent structure of a truncated P42345 in a complex with Q9BVC4 has provided a basic framework for understanding all of the phosphoinositide 3-kinase ( PI3K ) -related kinases ( PIKKs ) : P42345 , Q13315 , ATR , Q96Q15 , Q9Y4A5 and DNA-PK . The PIKK kinase domain is encircled by the FAT domain , a helical solenoid that is present in all PIKKs . PIKKs also have an extensive helical solenoid N-terminal to the FAT domain for which there is limited structural information . This N-terminal helical solenoid is essential for binding proteins that associate with the PIKKs to regulate their activity and cellular localization . Current researches on breast cancer epidemiology in Korea . As a cause of death in women , breast cancer ranks second to stomach cancer in Korea . Age-standardized mortality rates for breast cancer steadily increased during the 1980s and 1990s . There are big differences in the incidence rates for breast cancer compared with Western countries . Epidemiological features , trends in morbidity and mortality , various age-specific incidence curves , migrant study results , and analysis of the risk factors , however , suggest that the incidence of breast cancer might be further increasing in Korea . The key epidemiological hormonal risk factors for breast cancer are all explicable in terms of the estrogen augmented by progesterone hypothesis . These include older age , family history of breast cancer , early menarche , late menopause , late full-term pregnancy , and never a breast feeding . Both the establishment of high-risk groups and the estimation of lifetime risk are essential to develop a control strategy against breast cancer . Invasive ductal carcinoma is the most common histologic type of breast cancer in Korea , and the five-year survival rate has been estimated as 80-83 % . Recent studies on the identification of susceptibility factors such as genetic polymorphisms of P09488 /T1/P1 , P21964 , P05181 , P11511 , P05093 , P03372 , P18887 , O43542 , P43351 , TGF-alpha , P01375 , IL-1B , IL-1RN , P50613 etc. that predispose individuals to breast cancer by gene-environment or gene-gene interactions may possibly give further insight into both the etiology and the prevention of this malignancy . Comparison of genetic and epigenetic alterations at 11 tumor suppressor loci in pulmonary sclerosing hemangioma and adenocarcinoma . Pulmonary sclerosing hemangioma ( SH ) is an unusual tumor of pneumocytic origin . Morphologically , SH can mimic pulmonary adenocarcinomas . Here , the authors compared genetic and epigenetic aberrations in SH with those in pulmonary adenocarcinoma . Clinicopathologic characteristics , microsatellite alterations , and CpG island methylation were analyzed in pulmonary SHs ( n = 24 ) and adenocarcinomas ( n = 34 ) to compare their patterns of molecular abnormalities . SHs were also analyzed immunohistochemically to characterize the expression status of proteins involved in basic biologic processes . The clinical presentation of SH cases was generally benign . Both cell types of SH stained positive for thyroid transcription factor 1 ( Q15669 -1 ) , epithelial membrane antigen ( P15941 ) , β-catenin , P12830 , and vascular endothelial growth factor ( P15692 ) . Allelic imbalances in D3S1283 , D3S1234 , D3S1300 , D3S1285 , P04637 , D17S938 , and D9S179 were less frequent in SH than in adenocarcinoma ; rates of allelic imbalances in D20S170 and D21S1446 were not significantly different . In SH , CpG island methylation frequencies of p16(INK4a) ( 0.0 % ) and RASSF1A ( 12.5 % ) were significantly lower than those in adenocarcinoma ( 29.4 % and 38.2 % , respectively ) ; the frequencies of HOX D9 , D11 , and D13 gene methylation in SH were 37.5 % , 33.3 % , and 33.3 % , respectively . The results show that pulmonary SH and adenocarcinoma share similar genetic and epigenetic aberrations , but also exhibit significant differences , especially in tumor suppressor genes . Two-dimensional liquid crystalline growth within a phase-field-crystal model . By using a two-dimensional phase-field-crystal ( P27918 ) model , the liquid crystalline growth of the plastic triangular phase is simulated with emphasis on crystal shape and topological defect formation . The equilibrium shape of a plastic triangular crystal ( PTC ) grown from an isotropic phase is compared with that grown from a columnar or smectic-A ( Q13216 ) phase . While the shape of a PTC nucleus in the isotropic phase is almost identical to that of the classical P27918 model , the shape of a PTC nucleus in Q13216 is affected by the orientation of stripes in the Q13216 phase , and irregular hexagonal , elliptical , octagonal , and rectangular shapes are obtained . Concerning the dynamics of the growth process , we analyze the topological structure of the nematic order , which starts from nucleation of +1/2 and -1/2 disclination pairs at the PTC growth front and evolves into hexagonal cells consisting of +1 vortices surrounded by six satellite -1/2 disclinations . It is found that the orientational and the positional order do not evolve simultaneously ; the orientational order evolves behind the positional order , leading to a large transition zone , which can span over several lattice spacings . The effects of pertussis toxin on dopamine D2 and serotonin P08908 autoreceptor-mediated inhibition of neurotransmitter synthesis : relationship to receptor reserve . Irreversible inactivation of striatal D2 dopamine ( DA ) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ( EEDQ ) or inactivation of striatal guanine nucleotide binding proteins ( G proteins ) with pertussis toxin ( PT ) shifted the dose-response curve for N-n-propylnorapomorphine ( NPA ) -mediated inhibition of DB04699 ( Q9BVC4 ) -induced elevation of DB01235 ( DB01235 ) to the right , with a decrease in the maximum response . For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [ (+)-3-PPP ] , in contrast , there was little shift in the ED50 , after inactivation of either D2 receptors or G proteins . Completely analogous effects were found at the somatodendritic P08908 autoreceptor in the raphe nuclei , mediating inhibition of the synthesis of serotonin ( 5-HT ) ; the full agonist , 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) and the partial agonist , buspirone were utilized to inhibit the synthesis of 5-HT , as measured by changes in levels of L-5-hydroxytryptophan ( 5-HTP ) . Additionally , in both systems , combined treatment with pertussis toxin , followed by EEDQ , reduced the maximum effect , when compared to either agent alone but had little further effect on the ED50 . In systems exhibiting a large receptor reserve for agonists , such as those described above , the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . Tyrosine phosphorylation enhances P43351 -mediated annealing by modulating its DNA binding . P43351 protein has an important role in homology-directed DNA repair by mediating Q06609 nucleoprotein filament formation on single-stranded DNA ( ssDNA ) protected by replication protein-A ( RPA ) and annealing of RPA-coated ssDNA . In human , cellular response to DNA damage includes phosphorylation of P43351 by c- P00519 kinase at tyrosine 104 . To address how this phosphorylation modulates P43351 function , we used an amber suppressor technology to substitute tyrosine 104 with chemically stable phosphotyrosine analogue ( p-Carboxymethyl-L-phenylalanine , pCMF ) . The P43351 (Y104pCMF) retained ssDNA-binding activity characteristic of unmodified P43351 but showed lower affinity for double-stranded DNA ( dsDNA ) binding . Single-molecule analyses revealed that P43351 (Y104pCMF) specifically targets and wraps ssDNA . While P43351 (Y104pCMF) is confined to ssDNA region , unmodified P43351 readily diffuses into dsDNA region . The Y104pCMF substitution also increased the ssDNA annealing rate and allowed overcoming the inhibitory effect of dsDNA . We propose that phosphorylation at Y104 enhances ssDNA annealing activity of P43351 by attenuating dsDNA binding . Implications of phosphorylation-mediated activation of P43351 annealing activity are discussed . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . Loss of P38398 function increases the antitumor activity of cisplatin against human breast cancer xenografts in vivo . BACKGROUND : Previous reports suggested a central role of P38398 in DNA-damage repair mechanisms elicited by cell exposure to anti-tumor agents . Here we studied if P38398 -defective HCC1937 or P38398 -reconstituted HCC1937/(WT) P38398 human breast cancer xenografts ( HBCXs ) generated in SCID mice were differentially sensitive to cisplatin ( DB00515 ) in vivo and we investigated potential molecular correlates of this effect . RESULTS : DB00515 induced almost complete growth inhibition of P38398 -defective HBCXs , while P38398 -reconstituted HBCXs were only partially inhibited . Cell cycle analysis showed a significant S- and G(2)/M blockade in P38398 -defective as compared with parental P38398 -reconstituted cells . Comparative gene expression profiling of HCC1937 and HCC1937/(WT) P38398 showed upregulation of P43351 and Q13426 , whereas P07992 and P23921 were downregulated . Pathway finder analysis of gene arrays data indicated perturbations of major proliferation and survival pathways suggesting that P38398 is mostly involved in G(2)/M but also in G(1)/S-phase checkpoints as well as in several important signaling pathways , including IGF , P15692 , estrogen receptor , PI3K/AKT and P01133 . METHODS : HCC1937 or HCC1937/(WT) P38398 HBCXs were generated in SCID mice . Animals were then weekly treated with 5 mg/kg DB00515 i.p. or with vehicle for 4 w . Tumor volume and mice survival were evaluated . Tumors were retrieved from animals 12 hours after the last treatment with DB00515 or vehicle treatment and the cell suspension underwent cell cycle analysis . Differential gene expression and pathway modulation between HCC1937 and HCC1937/(WT) P38398 cells were also studied . CONCLUSION : Our data suggest that P38398 -defective in vivo HBCXs express a molecular scenario predictive of high sensitivity to platinum-derived compounds strongly supporting the rationale for prospective tailored clinical trials in hereditary breast cancer . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . O95760 augments DB05875 -induced P15692 secretion from human mast cells and is increased in psoriatic skin . The peptide DB05875 ( SP ) has been implicated in inflammatory conditions , such as psoriasis , where mast cells and P15692 are increased . A relationship between SP and P15692 has not been well studied , nor has any interaction with the proinflammatory cytokines , especially O95760 . Here we report that SP ( 0.1-10 microM ) induces gene expression and secretion of P15692 from human LAD2 mast cells and human umbilical core blood-derived cultured mast cells ( hCBMCs ) . This effect is significantly increased by coadministration of O95760 ( 5-100 ng/mL ) in both cell types . The effect of SP on P15692 release is inhibited by treatment with the P25103 antagonist 733,060 . SP rapidly increases cytosolic calcium , and so does O95760 to a smaller extent ; the addition of O95760 augments the calcium increase . SP-induced P15692 production involves calcium-dependent PKC isoforms , as well as the P29323 and JNK MAPKs . Gene expression of O95760 and histidine decarboxylase ( HDC ) , an indicator of mast cell presence/activation , is significantly increased in affected and unaffected ( at least 15 cm away from the lesion ) psoriatic skin , as compared with normal control skin . Immunohistochemistry indicates that O95760 is associated with endothelial cells in both the unaffected and affected sites , but is stronger and also associated with immune cells in the affected site . These results imply that functional interactions among SP , O95760 , and mast cells leading to P15692 release contribute to inflammatory conditions , such as the psoriasis , a nonallergic hyperproliferative skin inflammatory disorder with a neurogenic component . Acidic pH induces topoisomerase II-mediated DNA damage . Acidic pH plays an important role in various pathophysiological states and has been demonstrated to be carcinogenic in animal models . Recent studies have also implicated acidic pH in the development of preneoplastic Barrett 's esophagus in human . However , little is known about the molecular mechanism underlying acidic pH-induced carcinogenesis . In the current study , we show that acidic pH , like the topoisomerase II ( P11388 ) poison DB00773 ( demethylepipodophyllotoxin ethylidene-beta-D-glucoside ) , induces tumors in 9,10-dimethyl-1,2-benzanthracene(DMBA)-initiated mice . The following studies in tissue culture models have suggested that acidic pH acts like a P11388 poison to induce P11388 -mediated DNA damage : ( i ) acidic pH induces P11388 -dependent DNA damage signals as evidenced by up-regulation of p53 and DB00133 -139 phosphorylation of P16104 [ a substrate for ataxia telangiectasia mutated ( Q13315 ) Q13315 and Rad3-related ( ATR ) kinases ] ; ( ii ) acidic pH-induced cytotoxicity in tumor cells is reduced in P11388 -deficient cells ; ( iii ) acidic pH increases the mutation frequency of the hypoxanthine phosphoribosyl transferase ( P00492 ) gene in a P11388 -dependent manner ; and ( iv ) acidic pH induces reversible P11388 -mediated DNA strand breaks in vitro . We discuss the possibility that P11388 -mediated DNA damage may contribute to acidic pH-induced carcinogenesis . Metalloproteinases control brain inflammation induced by pertussis toxin in mice overexpressing the chemokine P13500 in the central nervous system . Inflammatory leukocytes infiltrate the CNS parenchyma in neuroinflammation . This involves cellular migration across various structures associated with the blood-brain barrier : the vascular endothelium , the glia limitans , and the perivascular space between them . Leukocytes accumulate spontaneously in the perivascular space in brains of transgenic ( Tg ) mice that overexpress P13500 under control of a CNS-specific promoter . The Tg mice show no clinical symptoms , even though leukocytes have crossed the endothelial basement membrane . Pertussis toxin ( PTx ) given i.p. induced encephalopathy and weight loss in Tg mice . We used flow cytometry , ultra-small superparamagnetic iron oxide-enhanced magnetic resonance imaging , and immunofluorescent staining to show that encephalopathy involved leukocyte migration across the glia limitans into the brain parenchyma , identifying this as the critical step in inducing clinical symptoms . Metalloproteinase ( MPs ) enzymes are implicated in leukocyte infiltration in neuroinflammation . Unmanipulated Tg mice had elevated expression of tissue inhibitor of metalloproteinase-1 , matrix metalloproteinase ( MMP ) -10 , and -12 mRNA in the brain . PTx further induced expression of tissue inhibitor of metalloproteinase-1 , metalloproteinase disintegrins-12 , P22894 , and -10 in brains of Tg mice . Levels of the microglial-associated MP P51511 were not affected in control or PTx-treated Tg mice . PTx also up-regulated expression of proinflammatory cytokines IL-1beta and P01375 mRNA in Tg CNS . Weight loss and parenchymal infiltration , but not perivascular accumulation , were significantly inhibited by the broad-spectrum MP inhibitor BB-94/ DB03880 . Our finding that MPs mediate PTx-induced parenchymal infiltration to the chemokine-overexpressing CNS has relevance for the pathogenesis of human diseases involving CNS inflammation , such as multiple sclerosis . GBM secretome induces transient transformation of human neural precursor cells . Glioblastoma ( GBM ) is the most aggressive primary brain tumor in humans , with a uniformly poor prognosis . The tumor microenvironment is composed of both supportive cellular substrates and exogenous factors . We hypothesize that exogenous factors secreted by brain tumor initiating cells ( BTICs ) could predispose normal neural precursor cells ( NPCs ) to transformation . When NPCs are grown in GBM-conditioned media , and designated as " tumor-conditioned NPCs " ( tcNPCs ) , they become highly proliferative and exhibit increased stem cell self-renewal , or the unique ability of stem cells to asymmetrically generate another stem cell and a daughter cell . tcNPCs also show an increased transcript level of stem cell markers such as CD133 and ALDH and growth factor receptors such as P17948 , P35968 , P00533 and PDGFRα . Media analysis by ELISA of GBM-conditioned media reveals an elevated secretion of growth factors such as P01133 , P15692 and PDGF-AA when compared to normal neural stem cell-conditioned media . We also demonstrate that tcNPCs require prolonged or continuous exposure to the GBM secretome in vitro to retain GBM BTIC characteristics . Our in vivo studies reveal that tcNPCs are unable to form tumors , confirming that irreversible transformation events may require sustained or prolonged presence of the GBM secretome . Analysis of GBM-conditioned media by mass spectrometry reveals the presence of secreted proteins Chitinase-3-like 1 ( P36222 ) and H2A histone family member P16104 . Collectively , our data suggest that GBM-secreted factors are capable of transiently altering normal NPCs , although for retention of the transformed phenotype , sustained or prolonged secretome exposure or additional transformation events are likely necessary . P00797 angiotensin system modulates P42345 pathway through AT2R in HIVAN . P42345 ( P42345 ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 and p70S6K . DB09026 , a renin inhibitor attenuated phosphorylation of both P42345 and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 ) phosphorylation of p70S6K in a dose dependent manner . HIV/ P04629 also displayed enhanced phosphorylation of both P42345 and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 and p70S6K . These findings indicate that HIV stimulates P42345 pathway in HIVAN through the activation of renin angiotensin system via AT2R . Assessing ' radiosensitivity ' with kinetic profiles of γ- P16104 , Q12888 and P38398 foci . BACKGROUND AND PURPOSE : DNA repair assays to identify radiosensitive patients have had limited clinical implementation due to long turn-around times or limited specificity . This study evaluates γ- P16104 -Irradiation Induced Foci ( IRIF ) kinetics as a more rapid surrogate for the ' gold standard ' colony survival assay ( Q13216 ) using several known DNA repair disorders as reference models . MATERIALS AND METHODS : Radiosensitive cells of known and unknown etiology were studied . γ- P16104 -IRIFs were quantified over 24 h , and the curves were fitted by combining logarithmic growth and exponential decay functions . Fitted values that differed from radionormal controls were considered aberrant and compared to Q13216 results . RESULTS : We observed 87 % agreement of IRIF data with the Q13216 for the 14 samples tested . Analysis of γ- P16104 -IRIF kinetics for known repair disorders indicated similarities between an Q8IYW5 (-/-) cell line and an RS cell of unknown etiology . These cell lines were further characterized by a reduction in P38398 -IRIF formation and G2/M checkpoint activation . CONCLUSIONS : γ- P16104 -IRIF kinetics showed high concordance with the Q13216 in RS populations demonstrating its potential as a more rapid surrogate assay . This method provides a means to globally identify defective DNA repair pathways in RS cells of unknown etiology through comparison with known DNA repair defects . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Mitoxantrone inhibits HIF-1α expression in a topoisomerase II-independent pathway . PURPOSE : Solid tumors encounter a growth-limiting hypoxic microenvironment as they develop . Hypoxia-inducible factors ( HIF ) play important roles in hypoxia-associated tumor development and therapeutic resistance . Targeting the HIF pathway ( especially HIF-1α ) represents a promising cancer treatment strategy . Here , we report a novel class of HIF-1α inhibitors and the possible molecular basis of inhibition . EXPERIMENTAL DESIGN : We analyzed the inhibitory effects of clinically used topoisomerase II ( P11388 ) -targeting drugs on HIF-1α expression with a primary focus on mitoxantrone . The potential role of P11388 in mitoxantrone-inhibited HIF-1α expression was studied using pharmacologic inhibition , a knockdown approach , and P11388 mutant cells . Moreover , involvement of mitoxantrone in proteasome-mediated degradation , transcription , and translation of HIF-1α was examined . RESULTS : The P11388 -targeting mitoxantrone , but neither doxorubicin nor etoposide ( DB00773 ) , strongly inhibited HIF-1α expression under hypoxic conditions in a dose- and time-dependent manner . Surprisingly , the mitoxantrone-mediated inhibition of HIF-1α expression was largely independent of two P11388 isozymes , proteasomal degradation , and transcription . Furthermore , mitoxantrone inhibited HIF-1α expression and function in a similar fashion as cycloheximide , suggesting that mitoxantrone might inhibit HIF-1α via a blockage at its translation step . In vitro translation experiments using HIF-1α mRNA further confirmed inhibition of HIF-1α translation by mitoxantrone . Interestingly , levels of the polysome-bound HIF-1α and P15692 mRNA were elevated and decreased after mitoxantrone treatment , respectively . CONCLUSIONS : We have identified the P11388 -targeting compound , mitoxantrone , as an HIF-1α inhibitor possibly through a translation inhibition mechanism , suggesting the possibility of an additional anticancer activity for mitoxantrone . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT/ P31749 phosphorylation . DB01200 ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 , official symbol Q01959 ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract . Isothiocyanate iberin modulates phase II enzymes , posttranslational modification of histones and inhibits growth of Caco-2 cells by inducing apoptosis . The aim of presented study was to further investigate the concentration-dependent changes induced by isothiocyanate iberin ( IBN ) in human colon carcinoma Caco-2 cells . The concentrations of IBN below IC(50) value ( 18 microM , 72 h ) triggered the augmentation of mRNA levels for phase II detoxification P08263 and P22309 enzymes and antioxidant thioredoxin reductase 1 gene in cells treated for 24 h . In addition a significant increase of acetylated H4 histone was detected . The mRNA induction peaked at IC(50) value and returned to level of control cells at 40 microM concentration of IBN . The cell cycle changes , gamma- P16104 stainability and the increase of phospho-H3 mitotic marker were induced at concentrations above IC(50) value . Appearance of P08758 positive apoptotic cells and sub- P55008 fragmented DNA as well as decrease of mitochondrial transmembrane potential confirmed cytotoxic effect of IBN observed in MTT assay . The predominance of necrotic cells and profound positivity of gamma- P16104 took place at the highest concentration of IBN . Thus , IBN represents the effective member of natural chemopreventive isothiocyanate family with which apoptotic potential can by employed to eliminate tumor cells . Resistance of glioblastoma-initiating cells to radiation mediated by the tumor microenvironment can be abolished by inhibiting transforming growth factor-β . The poor prognosis of glioblastoma ( GBM ) routinely treated with ionizing radiation ( IR ) has been attributed to the relative radioresistance of glioma-initiating cells ( GIC ) . Other studies indicate that although GIC are sensitive , the response is mediated by undefined factors in the microenvironment . GBM produce abundant transforming growth factor-β ( TGF-β ) , a pleotropic cytokine that promotes effective DNA damage response . Consistent with this , radiation sensitivity , as measured by clonogenic assay of cultured murine ( GL261 ) and human ( U251 , U87MG ) glioma cell lines , increased by approximately 25 % when treated with LY364947 , a small-molecule inhibitor of TGF-β type I receptor kinase , before irradiation . Mice bearing GL261 flank tumors treated with 1D11 , a pan-isoform TGF-β neutralizing antibody , exhibited significantly increased tumor growth delay following IR . GL261 neurosphere cultures were used to evaluate GIC . LY364947 had no effect on the primary or secondary neurosphere-forming capacity . IR decreased primary neurosphere formation by 28 % , but did not reduce secondary neurosphere formation . In contrast , LY364947 treatment before IR decreased primary neurosphere formation by 75 % and secondary neurosphere formation by 68 % . Notably , GL261 neurospheres produced 3.7-fold more TGF-β per cell compared with conventional culture , suggesting that TGF-β production by GIC promotes effective DNA damage response and self-renewal , which creates microenvironment-mediated resistance . Consistent with this , LY364947 treatment in irradiated GL261 neurosphere-derived cells decreased DNA damage responses , P16104 and p53 phosphorylation , and induction of self-renewal signals , Notch1 and P61073 . These data motivate the use of TGF-β inhibitors with radiation to improve therapeutic response in patients with GBM . A transgenic platform for testing drugs intended for reversal of cardiac remodeling identifies a novel 11βHSD1 inhibitor rescuing hypertrophy independently of re-vascularization . RATIONALE : Rescuing adverse myocardial remodeling is an unmet clinical goal and , correspondingly , pharmacological means for its intended reversal are urgently needed . OBJECTIVES : To harness a newly-developed experimental model recapitulating progressive heart failure development for the discovery of new drugs capable of reversing adverse remodeling . METHODS AND RESULTS : A P15692 -based conditional transgenic system was employed in which an induced perfusion deficit and a resultant compromised cardiac function lead to progressive remodeling and eventually heart failure . Ability of candidate drugs administered at sequential remodeling stages to reverse hypertrophy , enlarged LV size and improve cardiac function was monitored . Arguing for clinical relevance of the experimental system , clinically-used drugs operating on the P00797 -Angiotensin- DB04630 -System ( RAAS ) , namely , the P12821 inhibitor Enalapril and the direct renin inhibitor Aliskerin fully reversed remodeling . Remodeling reversal by these drugs was not accompanied by neovascularization and reached a point-of-no-return . Similarly , the PPARγ agonist Pioglitazone was proven capable of reversing all aspects of cardiac remodeling without affecting the vasculature . Extending the arsenal of remodeling-reversing drugs to pathways other than RAAS , a specific inhibitor of 11β-hydroxy-steroid dehydrogenase type 1 ( 11β HSD1 ) , a key enzyme required for generating active glucocorticoids , fully rescued myocardial hypertrophy . This was associated with mitigating the hypertrophy-associated gene signature , including reversing the myosin heavy chain isoform switch but in a pattern distinguishable from that associated with neovascularization-induced reversal . CONCLUSIONS : A system was developed suitable for identifying novel remodeling-reversing drugs operating in different pathways and for gaining insights into their mechanisms of action , exemplified here by uncoupling their vascular affects . MicroRNA-302 replacement therapy sensitizes breast cancer cells to ionizing radiation . PURPOSE : Solid tumors can be resistant or develop resistance to radiotherapy . The purpose of this study is to explore whether microRNA-302 is involved in radioresistance and can be exploited as a sensitizer to enhance sensitivity of breast cancer cells to radiation therapy . METHODS : MiR-302 expression levels in radioresistant cell lines were analyzed in comparison with their parent cell lines . Furthermore , we investigated whether enforced expression of miR-302 sensitized radioresistant breast cancer cells to ionizing radiation in vitro and in vivo . RESULTS : MiR-302 was downregulated in irradiated breast cancer cells . Additionally , the expression levels of miR-302a were inversely correlated with those of P31749 and P43351 , two critical regulators of radioresistance . More promisingly , miR-302a sensitized radioresistant breast cancer cells to radiation therapy in vitro and in vivo and reduced the expression of P31749 and P43351 . CONCLUSION : Our findings demonstrated that decreased expression of miR-302 confers radioresistance and restoration of miR-302 baseline expression sensitizes breast cancer cells to radiotherapy . These data suggest that miR-302 is a potential sensitizer to radiotherapy . Modeling the neurovascular niche : murine strain differences mimic the range of responses to chronic hypoxia in the premature newborn . Preterm birth results in significant cognitive and motor disabilities , but recent evidence suggests that there is variable recovery over time . One possibility that may explain this variable recovery entails variable neurogenic responses in the subventricular zone ( SVZ ) following the period of chronic hypoxia experienced by these neonates . In this report , we have characterized the responses to chronic hypoxia of two mouse strains that represent a wide range of susceptibility to chronic hypoxia . We determined that C57BL/6 pups and neural progenitor cells ( NPCs ) derived from them exhibit a blunted response to hypoxic insult compared with CD-1 pups and NPCs . Specifically , C57BL/6 pups and NPCs exhibited blunted in vivo and in vitro proliferative and increased apoptotic responses to hypoxic insult . Additionally , C57BL/6 NPCs exhibited lower baseline levels and hypoxia-induced levels of selected transcription factors , growth factors , and receptors ( including HIF-1alpha , Q9GZT9 , P23560 , P15692 , P48061 , TrkB , Nrp-1 , P61073 , and NO ) that determine , in part , the responsiveness to chronic hypoxic insult compared with CD-1 pups and NPCs , providing insight into this important and timely problem in perinatology . Targeting phosphoinositide 3-kinase : moving towards therapy . Phosphoinositide 3-kinases ( PI3K ) orchestrate cell responses including mitogenic signaling , cell survival and growth , metabolic control , vesicular trafficking , degranulation , cytoskeletal rearrangement and migration . Deregulation of the PI3K pathway occurs by activating mutations in growth factor receptors or the P42336 locus coding for P42336 , by loss of function of the lipid phosphatase and tensin homolog deleted in chromosome ten ( P60484 /MMAC/TEP1 ) , by the up-regulation of protein kinase B ( P31749 /Akt ) , or the impairment of the tuberous sclerosis complex ( Q92574 /2 ) . All these events are linked to growth and proliferation , and have thus prompted a significant interest in the pharmaceutical targeting of the PI3K pathway in cancer . Genetic targeting of P48736 ( P48736 ) and O00329 ( O00329 ) in mice has underlined a central role of these PI3K isoforms in inflammation and allergy , as they modulate chemotaxis of leukocytes and degranulation in mast cells . Proof-of-concept molecules selective for P48736 have already successfully alleviated disease progress in murine models of rheumatoid arthritis and lupus erythematosus . As targeting PI3K moves forward to therapy of chronic , non-fatal disease , safety concerns for PI3K inhibitors increase . Many of the present inhibitor series interfere with target of rapamycin ( TOR ) , DNA-dependent protein kinase ( DNA-PK(cs) ) and activity of the ataxia telangiectasia mutated gene product ( Q13315 ) . Here we review the current disease-relevant knowledge for isoform-specific PI3K function in the above mentioned diseases , and review the progress of > 400 recent patents covering pharmaceutical targeting of PI3K . Currently , several drugs targeting the PI3K pathway have entered clinical trials ( phase I ) for solid tumors and suppression of tissue damage after myocardial infarction ( phases I,II ) . DB00184 activates P46937 through nAChRs mediated signaling in esophageal squamous cell cancer ( ESCC ) . Cigarette smoking is an established risk factor for esophageal cancers . P46937 ( P46937 ) , the key transcription factor of the mammalian Hippo pathway , has been reported to be an oncogenic factor for many cancers . In this study , we find nicotine administration can induce nuclear translocation and activation of P46937 in ESCC . Consistently , we observed nuclear translocation and activation of P46937 by knockdown of P32297 , which is a negative regulator of nicotine signaling in bronchial and esophageal cancer cells . DB00184 administration or P32297 depletion substantially increased proliferation and migration in esophageal cancer cells . Interestingly , we find that P46937 physically interacts with nAChRs , and nAChRs-signaling dissociates P46937 from its negative regulatory complex composed with α-catenin , β-catenin and 14-3-3 in the cytoplasm , leading to upregulation and nuclear translocation of P46937 . This process likely requires PKC activation , as PKC specific inhibitor Enzastaurin can block nicotine induced P46937 activation . In addition , we find nicotine signaling also inhibits the interaction of P46937 with Q9H3D4 , which contributes to the inhibitory effect of nicotine on apoptosis . Using immunohistochemistry analysis we observed upregulation of P46937 in a significant portion of esophageal cancer samples . Consistently , we have found a significant association between P46937 upregulation and cigarette smoking in the clinical esophageal cancer samples . Together , these findings suggest that the nicotine activated nAChRs signaling pathway which further activates P46937 plays an important role in the development of esophageal cancer , and this mechanism may be of a general significance for the carcinogenesis of smoking related cancers . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . 7,12-Dimethylbenz[a]anthracene exposure induces the DNA repair response in neonatal rat ovaries . 7,12-Dimethylbenz[a]anthracene ( DMBA ) destroys ovarian follicles at all stages of development . This study investigated DMBA-induced DNA double strand break ( DSB ) formation with subsequent activation of the ovarian DNA repair response in models of pre-antral or pre-ovulatory follicle loss . Postnatal day ( P01160 ) 4 Fisher 344 ( F344 ) rat ovaries were cultured for 4 days followed by single exposures of vehicle control ( 1 % DB01093 ) or DMBA ( 12.5 nM or 75 nM ) and maintained in culture for 4 or 8 days . Alternately , PND4 F344 rat ovaries were exposed to 1 μM DMBA at the start of culture for 2 days . Total RNA or protein was isolated , followed by qPCR or Western blotting to quantify mRNA or protein level , respectively . γ P16104 and phosphorylated Q13315 were localized and quantified using immunofluorescence staining . DMBA exposure increased caspase 3 and γ P16104 protein . Additionally , DMBA ( 12.5 nM and 1 μM ) increased levels of mRNA encoding Atm , Xrcc6 , Brca1 and Rad51 . In contrast , Parp1 mRNA was decreased on d4 and increased on d8 of DMBA exposure , while P09874 protein increased after 8 days of DMBA exposure . Total Q13315 increased in a concentration-dependent temporal pattern ( 75 nM d4 ; 12.5 nM d8 ) , while pATM was localized in large primary and secondary follicles and increased after 8 days of 75 nM DMBA exposure compared to both control and 12.5 nM DMBA . These findings support that , despite some concentration effects , DMBA induces ovarian DNA damage and that DNA repair mechanisms are induced as a potential mechanism to prevent follicle loss . Landscape of somatic mutations and clonal evolution in mantle cell lymphoma . Mantle cell lymphoma ( Q8WXI8 ) is an aggressive tumor , but a subset of patients may follow an indolent clinical course . To understand the mechanisms underlying this biological heterogeneity , we performed whole-genome and/or whole-exome sequencing on 29 Q8WXI8 cases and their respective matched normal DNA , as well as 6 Q8WXI8 cell lines . Recurrently mutated genes were investigated by targeted sequencing in an independent cohort of 172 Q8WXI8 patients . We identified 25 significantly mutated genes , including known drivers such as ataxia-telangectasia mutated ( Q13315 ) , cyclin D1 ( P24385 ) , and the tumor suppressor P04637 ; mutated genes encoding the anti-apoptotic protein Q13489 and O60603 ( O60603 ) ; and the chromatin modifiers O96028 , MLL2 , and Q02080 . We also found Q04721 mutations as an alternative phenomenon to P46531 mutations in aggressive tumors with a dismal prognosis . Analysis of two simultaneous or subsequent Q8WXI8 samples by whole-genome/whole-exome ( n = 8 ) or targeted ( n = 19 ) sequencing revealed subclonal heterogeneity at diagnosis in samples from different topographic sites and modulation of the initial mutational profile at the progression of the disease . Some mutations were predominantly clonal or subclonal , indicating an early or late event in tumor evolution , respectively . Our study identifies molecular mechanisms contributing to Q8WXI8 pathogenesis and offers potential targets for therapeutic intervention . Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 in the actions of a 5-HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression/antidepressant response ; and second , by examining the role of the P08908 receptor subtype in the regulation of P15692 , and the cellular localization of antidepressant regulation of P15692 expression . The results show that pharmacological inhibition of P15692 receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 -Flk-1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 receptors is sufficient to induce P15692 expression and that a P08908 antagonist blocks both the increase in P15692 and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 receptors located on neurons and endothelial cells . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment . Amsacrine and etoposide hypersensitivity of yeast cells overexpressing DNA topoisomerase II . Increasing the cellular concentration of DNA topoisomerase II in yeast by expressing constitutively a plasmid-borne P11388 gene encoding the enzyme greatly increases the sensitivity of the cells to amsacrine and etoposide ( DB00773 ) . This increased drug sensitivity at a higher intracellular DNA topoisomerase II level is observed in both P43351 + repair-proficient strains and rad52 mutants that are defective in the repair of double-stranded breaks . These results provide strong support of the hypothesis that the cellular target of these drugs is DNA topoisomerase II , and that these drugs kill cells by converting DNA topoisomerase II into a DNA damaging agent . Akt/ P31749 suppresses DNA damage processing and checkpoint activation in late G2 . Using chemical genetics to reversibly inhibit Cdk1 , we find that cells arrested in late G2 are unable to delay mitotic entry after irradiation . Late G2 cells detect DNA damage lesions and form gamma- P16104 foci but fail to activate Chk1 . This reflects a lack of DNA double-strand break processing because late G2 cells fail to recruit RPA ( replication protein A ) , ATR ( ataxia telangiectasia and Rad3 related ) , Rad51 , or Q99708 ( C-terminal interacting protein ) to sites of radiation-induced damage , events essential for both checkpoint activation and initiation of DNA repair by homologous recombination . Remarkably , inhibition of Akt/ P31749 ( protein kinase B ) restores DNA damage processing and Chk1 activation after irradiation in late G2 . These data demonstrate a previously unrecognized role for Akt in cell cycle regulation of DNA repair and checkpoint activation . Because Akt/ P31749 is frequently activated in many tumor types , these findings have important implications for the evolution and therapy of such cancers . Lymphagenesis correlates with expression of vascular endothelial growth factor-C in colorectal cancer . Lymphagenesis in gastrointestinal tumors is not well described . To clarify its presence and regulation , we assessed the microlymphatic count ( MLC ) in colorectal cancer patients . Lymphatic vessels were evaluated by enzyme-histochemistry for 5'-nucleotidase ( 5'-NA ) . Since vascular endothelial growth factor ( P15692 ) -C is reportedly associated with lymphagenesis , the expression of P49767 protein was immunohistochemically assessed by the catalyzed signal amplification ( Q13216 ) method . MLC of peritumoral lesions was significantly higher than that of non-cancer and intratumoral lesions ( p < 0.01 ) ; it increased where P49767 was highly expressed ( p < 0.01 ) and increased with the depth of invasion in peritumoral lesions . These results indicate significant findings at peritumoral lesion : that lymphagenesis may be elicited by tumor spread ; that P49767 expression is associated with lymphagenesis and is a potent factor stimulating lymphagenesis . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . The role of vascular endothelial growth factor and matrix metalloproteinases in canine lymphoma : in vivo and in vitro study . BACKGROUND : Canine lymphoma represents the most frequent haematopoietic cancer and it shares some similarities with human non-Hodgkin lymphoma . Matrix metalloproteinases ( MMPs ) and vascular endothelial growth factor ( P15692 ) play a coordinated role during invasion and proliferation of malignant cells ; however , little is known about their role in canine haematologic malignancies . The aim of this study was to investigate the mRNA and protein expression of P15692 and the most relevant MMPs in canine lymphoma . Lymph node aspirates from 26 B-cell and 21 T-cell lymphomas were collected . The protein expression levels of P14780 , P08253 and P15692 were evaluated by immunocytochemistry , and the mRNA levels of P08253 , P14780 , P50281 , P01033 , P16035 , O95980 , P15692 and P15692 -164 were measured using quantitative RT-PCR . RESULTS : P50281 , P01033 and O95980 mRNA levels were significantly higher in T-cell lymphomas than in B-cell lymphomas . Higher mRNA and protein levels of P14780 and P15692 were observed in T-cell lymphomas than in B-cell lymphomas and healthy control lymph nodes . A positive correlation was found between P14780 and P15692 in T-cell lymphomas . Moreover , P14780 , P50281 , P01033 and P15692 were expressed at the highest levels in high-grade T-cell lymphomas . CONCLUSIONS : This study provides new information on the expression of different MMPs and P15692 in canine lymphoma , suggesting a possible correlation between different MMPs and P15692 , immunophenotype and prognosis . The dynamic alterations of P16104 complex during DNA repair detected by a proteomic approach reveal the critical roles of Ca(2+)/calmodulin in the ionizing radiation-induced cell cycle arrest . By using DNA nuclease digestion and a quantitative " dual tagging " proteomic approach that integrated mass spectrometry , stable isotope labeling , and affinity purification , we studied the histone P16104 -associating protein complex in chromatin in mammalian cells in response to ionizing radiation ( IR ) . In the non-irradiated control cells , calmodulin ( P62158 ) and the transcription elongation factor facilitates chromatin transcription ( FACT ) were associated with P16104 . Thirty minutes after exposing cells to IR the P62158 and FACT complexes dissociated , whereas two DNA repair proteins , poly(ADP-ribose) polymerase-1 and DEAH box polypeptide 30 isoform 1 , interacted with P16104 . Two hours and 30 min after exposure , none of the above proteins were in the complex . H2B , nucleophosmin/B23 , and calreticulin were associated with P16104 in both non-irradiated and irradiated cells . The results suggest that the P16104 complex undergoes dynamic changes upon induction of DNA damage and during DNA repair . The genuine interactions between P16104 and H2B , nucleophosmin/B23 , calreticulin , poly(ADP-ribose) polymerase-1 , and P62158 under each condition were validated by immunoprecipitation/Western blotting and mammalian two-hybrid assays . Because multiple Ca(2+)-binding proteins were found in the P16104 complex , the roles of Ca(2+) were examined . The results indicate that Ca(2+)/ P62158 plays important roles in regulating IR-induced cell cycle arrest , possibly through mediating chromatin structure . The dataset presented here demonstrates that sensitive profiling of the dynamics of functional cellular protein-protein interactions can successfully lead to the dissection of important metabolic or signaling pathways .

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Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Sustained increase of PKA activity in the postcommissural putamen of dyskinetic monkeys . Levodopa-induced dyskinesias ( LID ) are a frequent complication of Parkinson 's disease pharmacotherapy that causes significant disability and narrows the therapeutic window . Pharmacological management of LID is challenging partly because the precise molecular mechanisms are not completely understood . Here , our aim was to determine molecular changes that could unveil targetable mechanisms underlying this drug complication . We examined the expression and downstream activity of dopamine receptors ( DR ) in the striatum of 1-methyl-4-phenyl-1,2,3,6 tetrahydropiridine ( MPTP ) -lesioned monkeys with and without DB01235 treatment . Four monkeys were made dyskinetic and other four received a shorter course of DB01235 and did not develop LID . Our results show that DB01235 treatment induces an increase in P14416 and P35462 expression in the postcommissural putamen , but only P35462 is correlated with the severity of LID . Dyskinetic monkeys show a hyperactivation of the canonical P21728 -signaling pathway , measured by an increased phosphorylation of protein kinase A ( PKA ) and its substrates , particularly Q9UD71 . In contrast , activation of the P14416 -signaling pathway , visible in the levels of Akt phosphorylated on Thr308 and GSK3β on Ser9 , is associated with DB01235 treatment , independently of the presence of dyskinesias . Our data clearly demonstrate that dyskinetic monkeys present a dysregulation of the P35462 receptor and the P21728 pathway with a sustained increase of PKA activity in the postcommissural putamen . Importantly , we found that all signaling changes related to long-term DB01235 administration are exquisitely restricted to the postcommissural putamen , which may be related to the recurrent failure of pharmacological approaches . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Colocalization of somatostatin receptors with Q9UD71 in cortex and striatum of rat brain . Somatostatin ( P61278 ) -positive medium-sized aspiny interneurons are selectively spared in excitotoxicity . The biological effects of P61278 are mediated via five different receptors , namely somatostatin receptor (SSTR)1-5 ; however , SSTR subtype spared in excitotoxicity and involved in neuroprotection is not known . Dopamine- and DB02527 -regulated phosphoprotein ( Q9UD71 ) is predominantly expressed in medium-sized projection neurons that are most vulnerable in excitotoxicity . In the present study , we determined the colocalization of P61278 and SSTRs with Q9UD71 in rat brain cortical and striatal regions using immunofluorescence immunohistochemistry . We also determined the expression of Q9UD71 in P30872 -5 immunoprecipitate prepared from cortex and striatum . P61278 -positive neurons in cortex and striatum are devoid of colocalization with Q9UD71 . However , in cortical and striatal brain regions , three different neuronal populations either expressing SSTRs and Q9UD71 alone or displaying colocalization were identified . Quantitative analysis reveals that in cortex and striatum , P30872 and 5 are most predominant receptor subtypes colocalized with Q9UD71 followed by P31391 , 2 , and 3 in cortex whereas P30874 , 4 , and 3 in striatum . Importantly , Q9UD71 is expressed in P30872 -5 immunoprecipitate prepared from cortex and striatum . Taken together , these results provide the first evidence that the SSTR-positive neurons lacking colocalization with Q9UD71 might be spared in excitotoxicity . The A1 allele of the human D2 dopamine receptor gene is associated with increased activity of striatal L-amino acid decarboxylase in healthy subjects . The A1 allele of the TaqI restriction fragment length polymorphism ( RFLP ) of the human dopamine D2 receptor gene ( P14416 ) is associated with a low density of D2 dopamine receptors in the striatum . Because of the important role of D2 autoreceptors in regulating dopamine synthesis , we aimed to examine whether subjects with the A1 allele have altered presynaptic dopamine function in the brain . We also studied the effects of two other P14416 polymorphisms , C957 T and -- 141C Ins/Del , which have been suggested to affect D2 receptor levels in brain . The relationships between the TaqIA RFLP , C957 T and -- 141C Ins/Del polymorphisms and striatal dopamine synthesis in 33 healthy Finnish volunteers were studied using positron emission tomography and [18F]fluorodopa ( [18F]FDOPA ) , a radiolabelled analog of the dopamine precursor DB01235 . Heterozygous carriers of the A1 allele ( A1/A2 ; 10 subjects ) had significantly higher ( 18 % ) [18F]FDOPA uptake in the putamen than subjects without the A1 allele ( A2/A2 ; 23 subjects ) . C957 T and -- 141C Ins/Del polymorphisms did not significantly affect [18F]FDOPA Ki values . These results demonstrate that the A1 allele of P14416 gene is associated with increased striatal activity of aromatic L-amino acid decarboxylase , the final enzyme in the biosynthesis of dopamine and the rate-limiting enzyme for trace amine ( e.g. beta-phenylethylamine ) synthesis . The finding can be explained by lower D2 receptor expression leading to decreased autoreceptor function , and suggests that dopamine and/or trace amine synthesis rate is increased in the brains of A1 allele carriers . Radiation hybrid map of 13 loci on the long arm of chromosome 5 . Radiation hybrid mapping was used in conjunction with a natural deletion mapping panel to predict the order of and distance between 13 loci in the distal portion of the long arm of human chromosome 5 . A panel of irradiation hybrids containing fragments of 5q was generated from an P00492 + Chinese hamster-human cell hybrid containing a derivative chromosome 5 [ der(5)t(4;5) ( 5qter ---- 5p15.1::4p15.1 ---- 4pter ) ] as its only human DNA . One hundred nine radiation hybrids containing human DNA were screened with polymerase chain reaction primer sets representing nine genes encoding growth factors , growth factor receptors , or hormone receptors ( P08700 , P05112 , P05113 , P07333 , P05230 , P07550 , GRL , P14867 , and P21728 ) as well as four other loci ( P16591 , P09486 , P62263 , and P08571 ) to generate a radiation hybrid map of the area 5q21-q35 . A physical map predicting the order of and distance between the 13 loci was constructed based on segregation of the 13 loci in hybrid clones . The radiation hybrid panel will be useful as a mapping tool for determining the location and order of other genes and polymorphic loci in this region as well as for generating new DNA probes from specific regions . Systematic meta-analyses and field synopsis of genetic association studies in schizophrenia : the SzGene database . In an effort to pinpoint potential genetic risk factors for schizophrenia , research groups worldwide have published over 1,000 genetic association studies with largely inconsistent results . To facilitate the interpretation of these findings , we have created a regularly updated online database of all published genetic association studies for schizophrenia ( ' SzGene ' ) . For all polymorphisms having genotype data available in at least four independent case-control samples , we systematically carried out random-effects meta-analyses using allelic contrasts . Across 118 meta-analyses , a total of 24 genetic variants in 16 different genes ( P02649 , P21964 , DAO , P21728 , P14416 , P21917 , Q96EV8 , P47870 , Q13224 , HP , P01584 , P42898 , O75051 , P31645 , P04637 and P17752 ) showed nominally significant effects with average summary odds ratios of approximately 1.23 . Seven of these variants had not been previously meta-analyzed . According to recently proposed criteria for the assessment of cumulative evidence in genetic association studies , four of the significant results can be characterized as showing ' strong ' epidemiological credibility . Our project represents the first comprehensive online resource for systematically synthesized and graded evidence of genetic association studies in schizophrenia . As such , it could serve as a model for field synopses of genetic associations in other common and genetically complex disorders . Dopamine receptor agonists reverse behavioral abnormalities of alpha-synuclein transgenic mouse , a new model of Parkinson 's disease . Parkinson 's disease ( PD ) is characterized by loss of nigral dopaminergic ( DAergic ) neurons and presence of Lewy bodies , whose major component is alpha-synuclein . We had previously generated transgenic mice termed Syn130m that express truncated human alpha-synuclein ( amino acid residues 1-130 ) in DAergic neurons . Syn130m mice showed significant loss of DAergic neurons in the substantia nigra pars compacta . Subsequently , the striatal DA level and spontaneous locomotor activity of the mice were decreased significantly . In the present study , we investigated behavioral responses of Syn130m mice to DB01235 and DA receptor agonists . Administration of DB01235 dose dependently ameliorated the reduction of spontaneous locomotor activity of Syn130m mice . Similarly , D(2) agonists , quinpirole and talipexole , and a D1/D2 agonist , pergolide , were effective against the reduction . Syn130m mice also showed significant reduction in exploratory behavior compared with non-Tg littermates when they were placed in a novel environment , but this abnormality was ameliorated by treatment with pergolide . These results strongly suggest that the behavioral abnormalities of Syn130m mice were caused by low striatal DA content . On the other hand , the expression of postsynaptic D(2)-like receptors ( P14416 ) in the striatum was not increased in Syn130m mice , although the low striatal DA level is known to induce compensatory expression of P14416 . Because the abnormalities could be rectified by treatment with DA receptor agonists , it is likely that Syn130m mice provide a useful tool to explore therapeutic possibilities for PD as a new animal model of the disease . Stimulatory Effects of Peroxisome Proliferator-Activated Receptor-gamma on Fcgamma Receptor-Mediated Phagocytosis by Alveolar Macrophages . Alveolar macrophages abundantly express P37231 , with both natural and synthetic agonists maintaining the cell in a quiescent state hyporesponsive to antigen stimulation . Conversely , agonists upregulate expression and function of the cell-surface receptor P16671 , which mediates phagocytosis of lipids , apoptotic neutrophils , and other unopsonized materials . These effects led us to investigate the actions of P37231 agonists on the Fcgamma receptor , which mediates phagocytosis of particles opsonized by binding of immunoglobulin G antibodies . We found that troglitazone , rosiglitazone , and 15-deoxy-Delta12,14-prostaglandin J2 increase the ability of alveolar , but not peritoneal , macrophages to carry out phagocytosis mediated by the Fcgamma receptor . Receptor expression was not altered but activation of the downstream signaling proteins Syk , P27361 , and P28482 was observed . Although it was previously known that P37231 ligands stimulate phagocytosis of unopsonized materials , this is the first demonstration that they stimulate phagocytosis of opsonized materials as well . Stimulation of cyclic AMP formation in the circular smooth muscle of human colon by activation of Q13639 -like receptors . 5-HT stimulated cyclic AMP generation in human colonic circular smooth muscle in a concentration-dependent fashion ( EC50 = 229.1 nM ) . DAU 6236 also increased cyclic AMP formation and was a partial agonist relative to 5-HT . GR 113808 inhibited the cyclic AMP formation induced by 5-HT with a -log Ki value of 9.1 and an apparent pA2 value of 9.2 . DB00904 and methysergide failed to inhibit cyclic AMP formation induced by 5-HT . These results indicate that the Q13639 receptors of human colonic circular muscle mediate relaxation and inhibition of spontaneous contractions via formation of cyclic AMP . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . Blockade of globus pallidus adenosine A(2A) receptors displays antiparkinsonian activity in 6-hydroxydopamine-lesioned rats treated with D(1) or P14416 agonists . We have recently demonstrated how antagonism of adenosine A(2A) receptors within the globus pallidus ( GP ) ipsilateral to dopaminergic denervation potentiates contralateral rotational behavior induced by the dopamine precursor DB01235 in 6-hydroxydopamine-lesioned hemiparkinsonian rats . To further characterize the influence of pallidal A(2A) receptor blockade on the motor stimulant effects elicited by dopamine receptor activation , hemiparkinsonian rats were infused with the water-soluble A(2A) antagonist P35240 BT2 in the GP , alone or in combination with systemic administration of either SKF 38393 or quinpirole , to stimulate dopamine D(1) or D(2) receptors , respectively . P35240 BT2 alone ( 5 mug/1 mul ) neither altered motor behavior nor produced postural asymmetry . In contrast , the contralateral rotations elicited by SKF 38393 ( 1.5 mg/kg ) as well as quinpirole ( 0.05 mg/kg ) were potentiated by the concomitant intrapallidal infusion of P35240 BT2 . The results of this study demonstrate that blockade of pallidal A(2A) receptors exerts a facilitatory influence on the motor effects produced by the selective stimulation of either D(1) or D(2) dopamine receptors in hemiparkinsonian rats and suggest an involvement of GP in the antiparkinsonian activity of A(2A) receptor antagonists . Genetics of Alzheimer 's disease . A rapidly evolving field . Genetic factors have a variable impact on Alzheimer 's Disease ( AD ) , ranging from familial forms that are transmitted in an autosomal dominant fashion to sporadic AD , where a polygenic component is present . Most genes conferring susceptibility to AD are related to amyloid-beta deposition ( P05067 ; P49768 ; PS2 ; P02649 ; P01034 ; ubiquilin-1 ) , oxidative stress ( NOS2 ; NOS3 ) and inflammatory response ( P01583 ; P01584 ; P05231 ; P01375 ) . Genome-wide analyses , transcriptomics and proteomics approaches have pointed also to proapoptotic genes as increasing AD liability . Depression and psychotic symptoms that occur in a large proportion of AD patients have been associated with monoamine genes coding for metabolic enzymes ( P21964 ) , transporters ( 5-HTTLPR ) and receptors ( P21728 ; P35462 ) . Genetic testing may be useful to confirm the diagnosis of AD in individuals with clinical signs of dementia , while it is generally not recommended as a predictive testing for AD in asymptomatic individuals . Drugs currently in use to treat AD are effective in only 20 % of patients ; their therapeutic effect is predominantly under genetic control ( O43174 gene ; P02649 ) . Environmental factors have been shown to moderate the effects of genes on psychiatric disorders such as depression , schizophrenia and ADHD . The study of gene-environment interactions in AD , that are still poorly understood , is essential to predict disease-risk in asymptomatic individuals . Genomics will provide a dynamic picture of biological processes in AD and new targets for the forthcoming anti-AD drugs . Buspirone anti-dyskinetic effect is correlated with temporal normalization of dysregulated striatal P21728 signalling in DB01235 -treated rats . Dopamine replacement with l-DOPA is the most effective therapy in Parkinson 's disease . However , with chronic treatment , half of the patients develop an abnormal motor response including dyskinesias . The specific molecular mechanisms underlying dyskinesias are not fully understood . In this study , we used a well-characterized animal model to first establish the molecular differences between rats that did and did not develop dyskinesias . We then investigated the molecular substrates implicated in the anti-dyskinetic effect of buspirone , a 5HT1A partial agonist . Striatal protein expression profile of dyskinetic animals revealed increased levels of the dopamine receptor (DR)D3 , ΔFosB and phospho (p)CREB , as well as an over-activation of the P21728 signalling pathway , reflected by elevated ratios of phosphorylated Q9UD71 and P28482 . Buspirone reduced the abnormal involuntary motor response in dyskinetic rats in a dose-dependent fashion . Buspirone ( 4 mg/kg ) dramatically reduced the presence and severity of dyskinesias ( by 83 % ) and normalized Q9UD71 and P28482 phosphorylation ratios , while the increases in P35462 , ΔFosB and pCREB observed in dyskinetic rats were not modified . Pharmacological experiments combining buspirone with 5HT1A and P35462 antagonists confirmed that normalization of both pDARPP32 and pERK2 is required , but not sufficient , for blocking dyskinesias . The correlation between pDARPP32 ratio and dyskinesias was significant but not strong , pointing to the involvement of convergent factors and signalling pathways . Our results suggest that in dyskinetic rats P35462 striatal over-expression could be instrumental in the activation of P21728 -downstream signalling and demonstrate that the anti-dyskinetic effect of buspirone in this model is correlated with P21728 pathway normalization . Filamin A in somatostatin and dopamine receptor regulation in pituitary and the role of DB02527 /PKA dependent phosphorylation . Molecular mechanisms underlying resistance of pituitary tumors to somatostatin ( SS ) and dopamine ( DA ) analogues treatment are not completely understood . Resistance has been associated with defective expression of functional somatostatin and dopamine receptors P30874 , P35346 , and P14416 , respectively . Recently , a role of cytoskeleton protein filamin A ( P21333 ) in P14416 and SSTR receptors expression and signaling in PRL- and GH-secreting tumors , respectively , has been demonstrated , first revealing a link between P21333 expression and responsiveness of pituitary tumors to pharmacological therapy . No molecular events underlying the reduction of P21333 levels in resistant tumors have been so far identified . P21333 can be phosphorylated by PKA on Ser2152 , with increased P21333 resistance to cleavage by calpain and conformational changes affecting P21333 regions involved in P30874 and P14416 binding and signal transduction . In this respect , the effect of DB02527 /PKA pathway in the regulation of P21333 stability and/or function by modulating its phosphorylation status could assume particular importance in pituitary , where DB02527 cascade plays a crucial role in pituitary cell functions and tumorigenesis . This review will discuss the role of P21333 in the regulation of the main GPCRs target of pharmacological treatment of pituitary tumors , that is , P30874 and P14416 , focusing on the effects of DB02527 /PKA-mediated P21333 phosphorylation on P21333 biological functions . p38 mitogen-activated protein kinase mediates signal integration of TCR/ P10747 costimulation in primary murine T cells . Optimal T cell activation requires two signals , one generated by TCR and another by the P10747 costimulatory receptor . In this study , we investigated the regulation of costimulation-induced mitogen-activated protein kinase ( MAPK ) activation in primary mouse T cells . In contrast to that reported for human Jurkat T cells , we found that p38 MAPK , but not Jun NH2-terminal kinase ( JNK ) , is weakly activated upon stimulation with either anti-CD3 or anti- P10747 in murine thymocytes and splenic T cells . However , p38 MAPK is activated strongly and synergistically by either CD3/ P10747 coligation or PMA/Ca2+ ionophore stimulation , which mimics TCR-CD3/ P10747 -mediated signaling . Activation of p38 MAPK correlates closely with the stimulation of T cell proliferation . In contrast , PMA-induced JNK activation is inhibited by Ca2+ ionophore . T cell proliferation and production of P60568 , P05112 , and P01579 induced by both CD3 and CD3/ P10747 ligation and the nuclear expression of the c-Jun and P39905 -2 proteins are each blocked by the p38 MAPK inhibitor SB203580 . Our findings demonstrate that p38 P28482 ) plays an important role in signal integration during costimulation of primary mouse T cells , 2 ) may be involved in the induction of c-Jun activation and augmentation of AP-1 transcriptional activity , and 3 ) regulates whether T cells enter a state of functional unresponsiveness . Q9UD71 and inhibitor-1 are expressed in pancreatic beta-cells . P01308 secretion from pancreatic beta-cells has to be tightly regulated to ensure accurate glucose homeostasis . The capacity of beta-cells to respond to extracellular stimulation is determined by several signaling pathways . One important feature of these pathways is phosphorylation and subsequent dephosphorylation of a wide range of cellular substrates . Protein phosphatase 1 ( P50391 ) is a major eukaryotic serine/threonine protein phosphatase that controls a multitude of physiological processes . We have investigated the expression and cellular distribution of two endogenous inhibitors of P50391 activity in beta-cells . RT-PCR , Western blotting , and immunohistochemistry showed that Q9UD71 and inhibitor-1 are present in insulin-secreting endocrine beta-cells . Subcellular fractionation of mouse islets revealed that both P50391 inhibitors predominantly localized to cytosol-enriched fractions . Inhibitor-1 was also present in fractions containing plasma membrane-associated proteins . These data indicate a potential role for Q9UD71 and inhibitor-1 in the regulation of P50391 activity in pancreatic beta-cell stimulus-secretion coupling . Candidate gene studies of ADHD : a meta-analytic review . Quantitative genetic studies ( i.e. , twin and adoption studies ) suggest that genetic influences contribute substantially to the development of attention deficit hyperactivity disorder ( ADHD ) . Over the past 15 years , considerable efforts have been made to identify genes involved in the etiology of this disorder resulting in a large and often conflicting literature of candidate gene associations for ADHD . The first aim of the present study was to conduct a comprehensive meta-analytic review of this literature to determine which candidate genes show consistent evidence of association with childhood ADHD across studies . The second aim was to test for heterogeneity across studies in the effect sizes for each candidate gene as its presence might suggest moderating variables that could explain inconsistent results . Significant associations were identified for several candidate genes including Q01959 , P21917 , P21918 , P31645 , P28222 , and P60880 . Further , significant heterogeneity was observed for the associations between ADHD and Q01959 , P21917 , P21918 , P09172 , P08913 , P31645 , Q8IWU9 , P21397 , and P60880 , suggesting that future studies should explore potential moderators of these associations ( e.g. , ADHD subtype diagnoses , gender , exposure to environmental risk factors ) . We conclude with a discussion of these findings in relation to emerging themes relevant to future studies of the genetics of ADHD . DB09053 inhibits P11274 and NF-κB signaling and reduces tumor proliferation in tissue-resident cells of patients with CLL . Chronic lymphocytic leukemia ( CLL ) cells depend on microenvironmental factors for proliferation and survival . In particular , tissue-resident CLL cells show prominent activation of both B-cell receptor ( P11274 ) and NF-κB pathways . We evaluated the in vivo effects of ibrutinib , a Q06187 ( Q06187 ) inhibitor on tumor cell activation and proliferation in the blood , lymph node , and bone marrow of patients with CLL . Applying validated pathway-specific gene signatures , we detected a rapid and sustained downregulation of P11274 and NF-κB signaling in CLL cells from both the peripheral blood and tissue compartments during ibrutinib treatment . DB09053 reduced phosphorylation of PLCγ2 and P29323 and decreased nuclear protein expression of NF-κB p50 . DB09053 significantly decreased tumor proliferation and expression of surface activation markers Q07108 and P42081 , independent of prognostic factors such as IGHV mutational status , chromosome 17p deletion , or prior treatment history . Interestingly , stronger inhibition of P11274 signaling in lymph node resident CLL cells after one dose of ibrutinib was associated with a higher rate of nodal response at the end of cycle 2 . Together , these data validate on-target effects of Q06187 inhibition in the tissue compartments and demonstrate that ibrutinib effectively inhibits pathways that promote tumor cell activation and proliferation in vivo . This study is registered at www.clinicaltrials.gov as # NCT01500733 . The role of the subthalamic nucleus in DB01235 induced dyskinesia in 6-hydroxydopamine lesioned rats . DB01235 is the most effective treatment for Parkinson 's disease ( PD ) , but prolonged use leads to disabling motor complications including dyskinesia . Strong evidence supports a role of the subthalamic nucleus ( Q05639 ) in the pathophysiology of PD whereas its role in dyskinesia is a matter of controversy . Here , we investigated the involvement of Q05639 in dyskinesia , using single-unit extracellular recording , behavioural and molecular approaches in hemi-parkinsonian rats rendered dyskinetic by chronic DB01235 administration . Our results show that chronic DB01235 treatment does not modify the abnormal Q05639 activity induced by the 6-hydroxydopamine lesion of the nigrostriatal pathway in this model . Likewise , we observed a loss of Q05639 responsiveness to a single DB01235 dose both in lesioned and sham animals that received daily DB01235 treatment . We did not find any correlation between the abnormal involuntary movement ( AIM ) scores and the electrophysiological parameters of Q05639 neurons recorded 24 h or 20-120 min after the last DB01235 injection , except for the axial subscores . Nonetheless , unilateral chemical ablation of the Q05639 with ibotenic acid resulted in a reduction in global AIM scores and peak-severity of dyskinesia . In addition , Q05639 lesion decreased the anti-dyskinetogenic effect of buspirone in a reciprocal manner . Striatal protein expression was altered in dyskinetic animals with increases in ΔFosB , phosphoDARPP-32 , dopamine receptor ( DR ) D3 and P14416 / P21728 ratio . The Q05639 lesion attenuated the striatal molecular changes and normalized the P14416 / P21728 ratio . Taken together , our results show that the Q05639 plays a role , if modest , in the physiopathology of dyskinesias . [ Associations between six functional genes and schizophrenia ] . OBJECTIVE : To assess the associations between schizophrenia and six functional genes : dopamine D2 receptor gene ( P14416 ) , dopamine D4 receptor gene ( P21917 ) , 5-hydroxytryptamine 2A receptor gene ( 5- Q13049 ) , P50406 receptor gene ( P50406 ) , catechol-O-methyltransferase gene ( P21964 ) and dopamine transporter gene ( Q01959 ) . METHODS : With the techniques of Amp-RFLP and Amp- DB00712 , association analysis was made between schizophrenia and the six genes in 67 schizophrenic patients from Chinese Han population . RESULTS : ( 1 ) Neither genotypes nor alleles of P14416 , 5- Q13049 , P50406 and P21964 gene showed significant differences between patients and controls ( P > 0.05 ) . ( 2 ) Six repeats ( 6R ) in P21917 gene , the allele of 480 bp and the genotype of 480/520 in Q01959 gene were found to be of significant differences between the two groups ( P < 0.05 ) . ( 3 ) Only one negative association was observed between the 480 bp allele of Q01959 gene and schizophrenia ( OR=0.441 , 95 % CI:0.202-0.963 , Z=2.05 , P < 0.05 ) . CONCLUSION : The 480 bp allele of Q01959 gene is negatively associated with schizophrenia in Chinese Han population , which stands for the dopamine hypothesis of schizophrenia . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Dopamine receptor D4 polymorphism predicts the effect of DB01235 on gambling behavior . BACKGROUND : There is ample evidence that a subgroup of Parkinson 's disease patients who are treated with dopaminergic drugs develop certain behavioral addictions such as pathological gambling . The fact that only a subgroup of these patients develops pathological gambling suggests an interaction between dopaminergic drug treatment and individual susceptibility factors . These are potentially of genetic origin , since research in healthy subjects suggests that vulnerability for pathological gambling may be linked to variation in the dopamine receptor D4 ( P21917 ) gene . Using a pharmacogenetic approach , we investigated how variation in this gene modulates the impact of dopaminergic stimulation on gambling behavior in healthy subjects . METHODS : We administered 300 mg of L-dihydroxyphenylalanine ( DB01235 ) or placebo to 200 healthy male subjects who were all genotyped for their P21917 polymorphism . Subjects played a gambling task 60 minutes after DB01235 administration . RESULTS : Without considering genetic information , DB01235 administration did not lead to an increase in gambling propensity compared with placebo . As expected , however , an individual 's P21917 polymorphism accounted for variation in gambling behavior after the administration of DB01235 . Subjects who carry at least one copy of the 7-repeat allele showed an increased gambling propensity after dopaminergic stimulation . CONCLUSIONS : These findings demonstrate that genetic variation in the P21917 gene determines an individual 's gambling behavior in response to a dopaminergic drug challenge . They may have implications for the treatment of Parkinson 's disease patients by offering a genotype approach for determining individual susceptibilities for pathological gambling and may also afford insights into the vulnerability mechanisms underlying addictive behavior . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . The Q9Y275 /APRIL system : emerging functions beyond B cell biology and autoimmunity . The Q9Y275 system plays a key role in the development of autoimmunity , especially in systemic lupus erythematosus ( SLE ) . This often leads to the assumption that Q9Y275 is mostly a B cell factor with a specific role in autoimmunity . Focus on Q9Y275 and autoimmunity , driven by pharmaceutical successes with the recent approval of a novel targeted therapy DB08879 , has relegated other potential roles of Q9Y275 to the background . Far from being SLE-specific , the Q9Y275 system has a much broader relevance in infection , cancer and allergy . In this review , we provide the latest views on additional roles of the Q9Y275 system in health and diseases , as well as an update on Q9Y275 and autoimmunity , with particular focus on current clinical trials . N-acetylglucosamine prevents P01584 -mediated activation of human chondrocytes . DB01296 represents one of the most commonly used drugs to treat osteoarthritis . However , mechanisms of its antiarthritic activities are still poorly understood . The present study identifies a novel mechanism of glucosamine-mediated anti-inflammatory activity . It is shown that both glucosamine and N-acetylglucosamine inhibit IL-1beta- and P01375 -induced NO production in normal human articular chondrocytes . The effect of the sugars on NO production is specific , since several other monosaccharides , including glucose , glucuronic acid , and N-acetylmannosamine , do not express this activity . Furthermore , N-acetylglucosamine polymers , including the dimer and the trimer , also do not affect NO production . The observed suppression of IL-1beta-induced NO production is associated with inhibition of inducible NO synthase mRNA and protein expression . In addition , N-acetylglucosamine also suppresses the production of IL-1beta-induced cyclooxygenase-2 and P05231 . The constitutively expressed cyclooxygenase-1 , however , was not affected by the sugar . N-acetylglucosamine-mediated inhibition of the IL-1beta response of human chondrocytes was not associated with the decreased inhibition of the mitogen-activated protein kinases c-Jun N-terminal kinase , extracellular signal-related kinase , and p38 , nor with activation of the transcription factor NF-kappaB . In conclusion , these results demonstrate that N-acetylglucosamine expresses a unique range of activities and identifies a novel mechanism for the inhibition of inflammatory processes . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Biochemical and behavioral evidence for antidepressant-like effects of P50406 receptor stimulation . The primary action of several antidepressant treatments used in the clinic raises extracellular concentrations of serotonin ( 5-HT ) , which subsequently act on multiple 5-HT receptors . The present study examined whether P50406 receptors might be involved in the antidepressant-like effects mediated by enhanced neurotransmission at 5-HT synapses . A selective P50406 receptor antagonist , SB271046 , was evaluated for its ability to counteract fluoxetine-induced biochemical and behavioral responses in mice . In addition , biochemical and behavioral effects of the P50406 receptor agonist , 2-ethyl-5-methoxy-N,N-dimethyltryptamine ( EMDT ) , were assessed in mice to ascertain whether enhancement of P50406 receptor-mediated neurotransmission engenders antidepressant-like effects . SB271046 significantly counteracted the stimulatory actions of fluoxetine on cortical c-fos mRNA , phospho-Ser845-GluR1 , and in the tail suspension antidepressant assay , whereas it had no effect on these parameters by itself . EMDT increased the phosphorylation states of Thr34- Q9UD71 and Ser845-GluR1 , both in brain slices and in the intact brain , which were effects also seen with the antidepressant fluoxetine ; as with fluoxetine , these effects were demonstrated to be independent of D1 receptor stimulation . Systemic administration of EMDT increased c-fos mRNA expression in the striatum and cerebral cortex and reduced immobility in the tail suspension test . The antidepressant-like effects of EMDT in the tail suspension test were prevented by SB271046 . Our results indicate that P50406 receptor stimulation may be a mechanism initiating some of the biochemical and behavioral outcomes of 5-HT reuptake inhibitors , such as fluoxetine . These findings also indicate that selective P50406 receptor agonists may represent a novel antidepressant drug class . Comparison of rating scales used to evaluate DB01235 -induced dyskinesia in the 6-OHDA lesioned rat . Abnormal involuntary movement ( AIM ) rating scales are frequently used to study the mechanisms underlying DB01235 -induced dyskinesia ( LID ) in 6-OHDA lesioned rodents and the propensity of novel treatments for Parkinson 's disease to induce or alleviate similar abnormal behaviours . Despite the existence of at least one well validated method , other AIM scales are also in use . Moreover , there have been developments and variations in the original scales and their methods of use , without re-validation . In this study , 6-OHDA medial forebrain bundle lesioned Sprague-Dawley rats were treated with chronic DB01235 6 mg/kg/day for 5 weeks followed by 12 mg/kg/day for another 5 weeks . Rats were assessed weekly by simultaneous ratings on four published AIM and stereotypy scales with concurrent recording of rotation , over 3 hours following DB01235 injection . Three contemporary AIM scales have then been validated pharmacologically using agents that are known to reduce LID clinically and in primates ( amantadine ) or to interfere with the activity of DB01235 ( the D(1) and P14416 antagonists , P35240 -23390 and raclopride ) respectively . We also demonstrate that AIM , stereotypic and rotational behaviour are distinct motor dysfunctions induced by chronic and acute treatment of DB01235 , and should be assessed separately . The undertaking of assessments at multiple time points is essential especially when testing the efficacy of new potential anti-dyskinetic treatments . Importantly critical to all AIM and rotation testing is the internal validation of both the scale being used and the environment being used . P35462 stimulation underlies the development of DB01235 -induced dyskinesia in animal models of Parkinson 's disease . Development of DB01235 -induced dyskinesia ( LID ) remains a major problem in the long-term treatment of Parkinson 's disease ( PD ) . Sensitization to DB01235 correlates with ectopic expression of D3 dopamine receptors in the striatum , implicating D3 receptors in development of LID . We demonstrate that the selective D3 antagonist S33084 abolishes development of LID over 30 days in MPTP-lesioned marmosets without effecting the anti-parkinsonian actions of DB01235 . Furthermore , following a 14 day washout , when challenged with DB01235 in the absence of S33084 , these animals continued to exhibit reduced LID . In the 6-OHDA-lesioned rat , S33084 similarly attenuated development of behavioural sensitization to DB01235 . Additionally , DB01235 -induced elevations in striatal pre-proenkephalin-A ( PPE-A ) ( but not PPE-B , phospho[ DB00156 (34)] Q9UD71 , D1 , and D2 receptor mRNA or D3 receptor levels ) were reduced in S33084 treated animals . Our data suggest a role for D3 receptors in the development of LID and suggest that initiating DB01235 treatment with a D3 antagonist may reduce the development of LID in PD . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Nonlinkage of bipolar illness to tyrosine hydroxylase , tyrosinase , and D2 and D4 dopamine receptor genes on chromosome 11 . OBJECTIVE : Previous linkage and allelic association studies using DNA polymorphisms , cosegregation of cytogenetic abnormalities with psychiatric illness , and assignment of genes involved in neutotransmitter metabolism suggested that chromosome 11 may harbor a gene predisposing to bipolar illness . The authors examined linkage in the families of 14 probands with bipolar illness , with the candidate genes tyrosine hydroxylase ( TH ) , D4 dopamine receptor ( P21917 ) at 11p15 , tyrosinase ( P14679 ) at 11q14-q21 , and D2 dopamine receptor ( P14416 ) at 11q22-q23 , as well as with the c-Harvey-ras oncogene ( P01112 ) and insulin gene ( P01308 ) , both located at 11p15 , a region that previously showed linkage to bipolar illness . METHOD : The genetic data were analyzed with both lod score analysis ( parametric ) and affected-sib-pair analysis ( nonparametric ) ; both narrow and broad definitions of the clinical phenotype were used . Further influences of diagnostic uncertainties were accounted for by using diagnostic probability classes weighing the stability of each phenotype . RESULTS : Two-point linkage results excluded close linkage of bipolar illness to each candidate gene ; negative results were also obtained when the narrow definition of the clinical phenotype was used . Moreover , multipoint linkage analysis of P01112 and P01308 excluded the 11p15 region encompassing both P21917 and TH . In agreement with the negative linkage results , affected-sib-pair analysis did not show preferential sharing of marker alleles at any of the candidate genes . CONCLUSIONS : The negative results obtained under different genetic models exclude a frequent role for P21917 , TH , P14679 , and P14416 in the pathogenesis of bipolar illness . P06850 ( CRF ) over-expression down-regulates hippocampal dopamine receptor protein expression and CREB activation in mice . BACKGROUND : Stress results in hyperactivity of the hypothalamic-pituitary-adrenal ( Q9Y251 ) axis , characterized by increased central CRF activity , elevated circulating glucocorticoid levels , impaired glucocorticoid-mediated negative feedback and abnormal hippocampal functions , possibly contributing to the development of behavioral pathologies , such as depression . The hippocampus is critically involved in the control of the Q9Y251 axis as well as in explicit memory , contextual aspects of fear , organization of the behavioral response to environmental novelty and in habituation . We have previously shown that mice that over-express CRF in the brain exhibit impaired novelty detection and altered psychophysiological and behavioral habituation , functions linked to dopamine receptor-dependent hippocampal plasticity . OBJECTIVE AND METHODS : Therefore , the aim of the present study was to measure D1 and D2 dopamine receptor expression and related signaling , such as CREB and P29323 protein levels and phosphorylation , in the hippocampus and other brain regions of mice with post-natal CRF over-expression ( CRF-OE mice ) . RESULTS : We found a region-specific down-regulation of both D1 and D2 protein expression , without altered CRF receptor protein expression , in the hippocampus in CRF-OE mice . This was accompanied by an impaired phosphorylation of hippocampal CREB , but not P27361 and P28482 , in the same animals . CONCLUSIONS : These results suggest that post-natal onset CRF over-expression results in an impairment of dopamine signaling in the hippocampus , which may underlie cognitive and motivational aspects of stress-related , CRF-driven mood disorders . DB01235 -treatment in primates disrupts the expression of A(2A) adenosine-CB(1) cannabinoid- P14416 heteromers in the caudate nucleus . The molecular basis of priming for DB01235 -induced dyskinesias in Parkinson 's disease ( PD ) , which depends on the indirect pathway of motor control , is not known . In rodents , the indirect pathway contains striatopallidal GABAergic neurons that express heterotrimers composed of A(2A) adenosine , CB(1) cannabinoid and D(2) dopamine receptors that regulate dopaminergic neurotransmission . The present study was designed to investigate the expression of these heteromers in the striatum of a primate model of Parkinson 's disease and to determine whether their expression and pharmacological properties are altered upon DB01235 treatment . By using the recently developed in situ proximity ligation assay and by identification of a biochemical fingerprint , we discovered a regional distribution of A(2A)/CB(1) /D(2) receptor heteromers that predicts differential D(2)-mediated neurotransmission in the caudate-putamen of Macaca fascicularis . Whereas heteromers were abundant in the caudate nucleus of both naïve and MPTP-treated monkeys , DB01235 treatment blunted the biochemical fingerprint and led to weak heteromer expression . These findings constitute the first evidence of altered receptor heteromer expression in pathological conditions and suggest that drugs targeting A(2A)-CB(1) -D(2) receptor heteromers may be successful to either normalize basal ganglia output or prevent DB01235 -induced side effects . Task-dependent interactions between dopamine D2 receptor polymorphisms and DB01235 in patients with Parkinson 's disease . Variants in genes regulating dopamine transmission affect performance on tasks including working memory and executive function as well as temporal processing and sequence learning . In the current study , we determined whether a dopamine D2 receptor DNA sequence polymorphism interacts with DB01235 during motor tasks in patients with Parkinson 's disease ( PD ) . Forty-five PD patients were genotyped for the P14416 polymorphism ( rs 1076560 , G > T ) . Patients performed an explicit motor sequence learning task and the grooved pegboard test in both ON and OFF DB01235 states . For motor sequence learning , P14416 genotype mediated DB01235 effects such that DB01235 associated improvements were only observed in the minor T allele carriers ( associated with lower D2 receptor availability , t10=-2.71 , p=0.022 ) , whereas G homozygotes showed no performance change with DB01235 . For the grooved pegboard test , performance improved with DB01235 independent of patients ' P14416 genotype . Collectively these results demonstrate that common P14416 allelic differences found in the human population may explain how dopamine differentially contributes to performance across tasks and individuals . DB00104 is the favorable alternative for cisplatin resistance reversal of ovarian cancer in vitro and in nude mice in vivo . This study aimed to observe the effects of octreotide ( O75051 ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo . MTT method and flow cytometry were used to investigate the effect of cisplatin , O75051 or the combination of these two compounds on the proliferation and apoptosis of SKOV3- O60220 cells . The size and weight of xenograft tumors from the nude mice model were measured . Real-time PCR was used to detect the mRNA expression of P30874 , P08183 , Q92887 , Q86UG4 -pi and P00533 in SKOV3/ O60220 cells following the different treatment . At the concentration of 2.5-20 g/ml , O75051 significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3- O60220 cells ' response to cisplatin . Unchanged expression was found in P30874 on the SKOV3/ O60220 cell in vitro after O75051 treatment , but increased expression in vivo ( p < 0.05 ) . O75051 increased Q86UG4 -pi expression ( p < 0.05 ) and reduced Q92887 and P00533 expression ( p < 0.05 ) in a dose-dependent manner . The similar results were obtained in mice in vivo experiment , except the reduced expression of Q86UG4 -pi . It is suggested that O75051 could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface P30874 , and reverse cisplatin resistance through inhibition of Q92887 , P00533 , and even Q86UG4 -pi expressions . P04150 recruitment of histone deacetylase 2 inhibits interleukin-1beta-induced histone H4 acetylation on lysines 8 and 12 . We have investigated the ability of dexamethasone to regulate interleukin-1beta ( IL-1beta ) -induced gene expression , histone acetyltransferase ( O60235 ) and histone deacetylase ( HDAC ) activity . Low concentrations of dexamethasone ( 10(-10) M ) repress IL-1beta-stimulated granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) expression and fail to stimulate secretory leukocyte proteinase inhibitor expression . Dexamethasone ( 10(-7) M ) and IL-1beta ( 1 ng/ml ) both stimulated O60235 activity but showed a different pattern of histone H4 acetylation . Dexamethasone targeted lysines P13647 and P08779 , whereas IL-1beta targeted K8 and K12 . Low concentrations of dexamethasone ( 10(-10) M ) , which do not transactivate , repressed IL-1beta-stimulated K8 and K12 acetylation . Using chromatin immunoprecipitation assays , we show that dexamethasone inhibits IL-1beta-enhanced acetylated K8-associated GM- P04141 promoter enrichment in a concentration-dependent manner . Neither IL-1beta nor dexamethasone elicited any GM- P04141 promoter association at acetylated P13647 residues . Furthermore , we show that GR acts both as a direct inhibitor of CREB binding protein ( CBP ) -associated O60235 activity and also by recruiting Q92769 to the p65-CBP O60235 complex . This action does not involve de novo synthesis of HDAC protein or altered expression of CBP or p300/CBP-associated factor . This mechanism for glucocorticoid repression is novel and establishes that inhibition of histone acetylation is an additional level of control of inflammatory gene expression . This further suggests that pharmacological manipulation of of specific histone acetylation status is a potentially useful approach for the treatment of inflammatory diseases . P16150 - , but not P16150 + , P22301 -producing CD1dhiCD5+ B cells suppress type 1 immune responses during Chlamydia muridarum genital tract infection . Regulatory B ( Breg ) cells are known to modulate immune responses through predominantly interleukin-10 ( P22301 ) -dependent mechanisms and can be hypothetically divided into innate and adaptive subsets based on the nature of their activating signals . However , the specific role of different Breg subsets in modulating immune responses remains ambiguous . Here we have shown that Chlamydia induces P22301 -producing splenic B-cell populations consisting of P16150 (+) and P16150 (-) subsets of IgM(hi)IgD(lo) innate-like B ( ILB ) cells in vitro . While P16150 (+) P22301 -producing B cells displayed innate type features and were readily induced by Chlamydia via Toll-like-receptor ( TLR ) signaling , P16150 (-) P22301 -producing B cells required additional B-cell activating factor ( Q9Y275 ) -mediated signals from dendritic cells ( DCs ) for their differentiation and activation , thereby classifying them as adaptive type Bregs . Importantly , P16150 (-) , but not P16150 (+) , P22301 -producing ILB cells displayed bona fide Breg activity by potently suppressing interferon-γ ( IFN-γ ) production in vitro in an P22301 -dependent manner . Furthermore , a novel P16150 (-)CD1d(hi) P06127 (+) P22301 -producing Breg population was predominantly induced by Chlamydia genital infection in vivo . Correspondingly , mixed bone marrow chimeric mice with B-cell-specific P22301 deficiency exhibited significantly increased type 1 immune responses , decreased bacterial burden , and reduced oviduct pathology upon infection . Our data demonstrate for the first time a distinct role for P16150 (-)CD1d(hi) P06127 (+)-adaptive Bregs over P16150 (+) innate counterparts in controlling mucosal responses against intracellular bacterial infection . Severe acute respiratory syndrome coronavirus papain-like protease suppressed alpha interferon-induced responses through downregulation of extracellular signal-regulated kinase 1-mediated signalling pathways . Severe acute respiratory syndrome coronavirus ( P49591 -CoV ) papain-like protease ( PLpro ) , a deubiquitinating enzyme , reportedly blocks poly I : C-induced activation of interferon regulatory factor 3 and nuclear factor kappa B , reducing interferon ( IFN ) induction . This study investigated type I IFN antagonist mechanism of PLpro in human promonocytes . PLpro antagonized IFN-α-induced responses such as interferon-stimulated response element- and AP-1-driven promoter activation , protein kinase R , 2'-5'-oligoadenylate synthetase ( OAS ) , interleukin ( IL ) -6 and P10145 expression , and signal transducers and activators of transcription ( P35610 ) 1 ( Tyr701 ) , P42224 ( Ser727 ) and c-Jun phosphorylation . A proteomics approach demonstrated downregulation of extracellular signal-regulated kinase ( P29323 ) 1 and upregulation of ubiquitin-conjugating enzyme ( P0CG48 ) E2-25k as inhibitory mechanism of PLpro on IFN-α-induced responses . IFN-α treatment significantly induced mRNA expression of P0CG48 E2-25k , but not P27361 , causing time-dependent decrease of P27361 , but not P28482 , in PLpro-expressing cells . Poly-ubiquitination of P27361 showed a relationship between P27361 and ubiquitin proteasome signalling pathways associated with IFN antagonism by PLpro . Combination treatment of IFN-α and the proteasome inhibitor MG-132 showed a time-dependent restoration of P27361 protein levels and significant increase of P27361 , P42224 and c-Jun phosphorylation in PLpro-expressing cells . Importantly , PD098059 ( an P27361 /2 inhibitor ) treatment significantly reduced IFN-α-induced P27361 and P42224 phosphorylation , inhibiting IFN-α-induced expression of 2'-5'-OAS in vector control cells and PLpro-expressing cells . Overall results proved downregulation of P27361 by ubiquitin proteasomes and suppression of interaction between P27361 and P42224 as type I IFN antagonist function of P49591 -CoV PLpro . P21728 behavioral responsitivity following selective lesions of the striatal patch compartment during development . The behavioral effects of selective destruction of the dopamine ( DA ) input to the patch compartment of rat striatum early in development was investigated . Rat pups were given bilateral intrastriatal ( i.s. ) injections of the neurotoxin 6-hydroxydopamine ( 6-OHDA ) on day of birth ( P0 ) or postnatal day 1 ( P1 ) , which resulted in selective behavioral alterations following DA agonist treatment in adulthood . Neonatally-lesioned rats exhibited self-biting behavior following treatment with the DA precursor L-dihydroxyphenylalanine ( DB01235 ) . In response to treatment with the selective D1 agonist SKF38393 , there was an increased incidence of abnormal perioral movements . The cataleptogenic effects of the D1 antagonist SCH23390 and the D2 antagonist haloperidol were also studied . Neonatally-lesioned rats were significantly less cataleptic compared to control rats following D1 antagonist treatment , but not following D2 antagonist treatment . Autoradiographs of [3H]mazindol binding to DA uptake sites ( a measure of DA terminal density ) showed a ' patchy ' loss of approx . 40-50 % in striatal tissue sections derived from the i.s. lesioned rats . These data suggest that injections of 6-OHDA into the striatum during this early postnatal period cause a DA lesion that results in long-term effects on a D1 receptor system . Permanent neonatal diabetes mellitus in China . BACKGROUND : Permanent neonatal diabetes mellitus ( PNDM ) is a rare disease , which is defined as the onset of diabetes before the age of 6 months with persistence through life . Infants with Q14654 or Q09428 genetic mutations may respond to oral sulfonylurea therapy . Currently , there are limited studies about the genetic analysis and long-term follow-up of PNDM . CASE PRESENTATION : We report four cases of PNDM . None of the infants or their parents had P01308 , Q14654 , or Q09428 genetic mutations . One infant underwent continuous subcutaneous insulin infusion ( CSII ) and the other infants underwent multiple injections of insulin ( MII ) . In these infants , PNDM persisted from 35 months to 60 months of follow-up . Three infants maintained fairly stable blood sugar levels , and one infant had poor sugar control . CONCLUSIONS : We suggest that all of the infants with PNDM should undergo genetic evaluation . For infants without Q14654 and Q09428 genetic mutations , oral sulfonylurea should not be considered as treatment . CSII is a useful method for overcoming the difficulties of diabetes , and it may also improve the quality of life of both infants and their parents . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Neurogenetics of aggressive behavior : studies in primates . Aggressive behavior can have adaptive value in certain environmental contexts , but when extreme or executed inappropriately , can also lead to maladaptive outcomes . Neurogenetic studies performed in nonhuman primates have shown that genetic variation that impacts reward sensitivity , impulsivity , and anxiety can contribute to individual differences in aggressive behavior . Genetic polymorphisms in the coding or promoter regions of the Mu-Opioid Receptor ( P35372 ) , DB01285 Releasing Hormone ( P06850 ) , Monoamine Oxidase A ( P21397 ) , Dopamine D4 Receptor ( P21917 ) , and Serotonin Transporter ( P31645 ) genes have been shown to be functionally similar in humans and rhesus macaques and have been demonstrated to contribute to individual differences in aggression . This body of literature suggests mechanisms by which genetic variation that promotes aggressivity could simultaneously increase evolutionary success while making modern humans more vulnerable to psychopathology . Therapy with interferon-beta modulates endogenous catecholamines in lymphocytes of patients with multiple sclerosis . OBJECTIVE : To investigate the endogenous dopaminergic/adrenergic system of lymphocytes in multiple sclerosis ( MS ) patients during treatment with interferon ( IFN ) -beta . METHODS : Patients with relapsing-remitting MS undergoing IFN-beta treatment were prospectively studied during the first year of treatment . Circulating lymphocytes were obtained at baseline and after 1 , 3 , 6 and 12 months of treatment and assayed for catecholamine ( CA ) production and mRNA expression of tyrosine hydroxylase ( TH , the rate-limiting enzyme in the synthesis of CA ) , beta(2)-adrenoceptors ( AR ) and D2 , D3 and D5 dopaminergic receptors ( DR ) . RESULTS : In cells from patients treated with IFN-beta for 12 months the production of CA hugely increased and was less sensitive to P01579 -induced inhibition . Expression of mRNA for TH , beta(2)-AR and P21918 was already enhanced after 1 month and further increased up to 6-12 months of treatment . On the contrary , P14416 mRNA progressively decreased and P35462 mRNA did not significantly change over the whole study period . CONCLUSIONS : In MS patients IFN-beta treatment enhances the ability of lymphocytes to produce CA , and induces extensive modifications of both beta(2)-AR and DR-operated pathways . The clinical relevance of these effects deserves consideration . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Involvement of histone deacetylation in ras-induced down-regulation of the metastasis suppressor O95980 . O95980 is a membrane-anchored glycoprotein that may negatively regulate matrix metalloproteinase ( MMP ) activity and inhibit tumor metastasis . Previous study demonstrated that oncogenic ras inhibited O95980 expression via an Sp1 binding site in the O95980 promoter . In this study , we investigated the molecular mechanism by which ras inhibited O95980 expression . Co-transfection assay showed that Sp1 and Sp3 are transactivators , rather than repressors , for O95980 gene . So , we tested whether ras activation induced the binding of histone deacetylases ( HDACs ) to Sp1 to repress O95980 expression . Our data showed Sp1-associated Q13547 in cells was increased after ras induction . By using DNA affinity precipitation assay , we found that induction of oncogenic ras enhanced the binding of Q13547 to the DNA probe corresponding to the Sp1 site in the O95980 promoter . Additionally , a HDAC inhibitor trichostatin A ( P32119 ) potently antagonized the inhibitory action of ras on O95980 . The signaling pathway by which ras suppresses O95980 was also addressed . Induction of oncogenic ras activated extracellular signal-regulated kinase ( P29323 ) , but not c-Jun N-terminal kinase ( JNK ) and p38(HOG) kinase in 2-12 cells . Addition of PD98059 or overexpression of dominant-negative mutant of P28482 indeed reversed ras-mediated inhibition of O95980 promoter activity . Taken together , our results suggest that oncogenic ras represses O95980 expression via a histone deacetylation mechanism . Q92769 attenuates P50591 -induced apoptosis of pancreatic cancer cells . BACKGROUND : Pancreatic ductal adenocarcinoma ( PDAC ) is one of the most malignant tumors with a dismal prognosis and no effective conservative therapeutic strategies . Although it is demonstrated that histone deacetylases ( HDACs ) , especially the class I HDACs Q13547 , 2 and 3 are highly expressed in this disease , little is known about HDAC isoenzyme specific functions . RESULTS : Depletion of Q92769 , but not Q13547 , in the pancreatic cancer cell lines MiaPaCa2 and Panc1 resulted in a marked sensitization towards the tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) . Correspondingly , the more class I selective HDAC inhibitor ( HDACI ) valproic acid ( DB00313 ) synergized with P50591 to induce apoptosis of MiaPaCa2 and Panc1 cells . At the molecular level , an increased expression of the O00220 ( DR5 ) , accelerated processing of caspase 8 , pronounced cleavage of the Q7L3V2 Bid , and increased effector caspase activation was observed in Q92769 -depleted and P50591 -treated MiaPaCa2 cells . CONCLUSIONS : Our data characterize a novel Q92769 function in PDAC cells and point to a strategy to overcome P50591 resistance of PDAC cells , a prerequisite to succeed with a P50591 targeted therapy in clinical settings . The interleukin 10 promoter haplotype ACA and the long-form variant of the P21917 uVNTR polymorphism are associated with vulnerability to schizophrenia . A total of 934 patients with schizophrenia and 433 controls were genotyped for the interleukin-10 ( P22301 ) promoter and P21917 uVNTR polymorphisms . P21917 long-form variants ( namely , those with ≥5 repeats ) , homozygosity for the 4-repeat allele , and the P22301 haplotype ACA were associated with schizophrenia , respectively . No obvious interactions among the potential polymorphisms were found , which suggests that P22301 and P21917 confer vulnerability to schizophrenia independently . 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 . Dopamine-related genes and their relationships to monoamine metabolites in P04141 . Monoamine metabolite ( MM ) levels in lumbar cerebrospinal fluid ( P04141 ) are extensively used as indirect estimates of monoamine turnover in the brain . In this study we investigated genotypes for DNA polymorphisms in the D2 ( P14416 ) , D3 ( P35462 ) , and D4 ( P21917 ) dopamine receptor and tyrosine hydroxylase ( TH ) genes and their relationships to P04141 MM in healthy volunteers ( n = 66 ) . Concentrations of homovanillic acid ( HVA ) , 3-methoxy-4-hydroxyphenylglycol ( MHPG ) , and 5-hydroxyindoleacetic acid ( 5-HIAA ) were corrected for back length , a confounding variable . Corrected MM levels were not related to age , gender , height , weight heredity , season or atmospheric pressure at sampling . Individuals with specific P14416 and TH allele and genotype configurations significantly differed in HVA and MHPG concentrations . P35462 homo- and heterozygotic genotypes had significantly different P04141 5-HIAA levels . P21917 genotypes were not related to MM concentrations . The results suggest that specific P14416 , P35462 , and TH genotypes participate in the regulation of monoamine turnover in the central nervous system . Accordingly monoamine receptors and synthesizing enzyme genotypes appear to be variance factors influencing MM concentrations in P04141 . The relationships found in this study support MM concentrations as markers for monoamine transmission in the human brain . The P38936 codon 31*C- and P14416 codon 313*T-related genotypes/alleles , but not P18887 codon 399 , hOGG1 codon 326 , and P21728 -48 polymorphisms , are correlated with the presence of leiomyoma . OBJECTIVE : To investigate whether the gene polymorphisms for P38936 , X-ray repair cross-complementing group 1 ( P18887 ) , human 8-oxoguanine glycosylase 1 ( hOGG1 ) , and dopamine D1 and D2 receptors ( P21728 , -2 ) are associated with leiomyoma susceptibility . DESIGN : Prospective study . SETTING : Departments of gynecology and genetics in a medical center . PATIENT(S) : Women were divided into two groups : leiomyoma ( n = 120 ) and nonleiomyoma ( n = 112 ) . INTERVENTION(S) : The P38936 codon 31 , P18887 codon 399 , hOGG1 codon 326 , P21728 -48 , and P14416 codon 313 polymorphisms were genotyped by polymerase chain reaction with restriction enzyme digestions ( Blp I , MspI , Fnu4HI , Dde I , and NcoI , respectively ) . MAIN OUTCOME MEASURE(S) : Genotypes and allelic frequencies . RESULT(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyomas . The proportions of P38936 (*)CC/CA/AA and P14416 (*)CC/CT/TT in both groups were 27.5/68.3/4.2 % and 12.5/51.7/35.8 % ( leiomyoma ) ; and 14.3/51.8/33.9 % and 33.9/40.2/25.9 % ( nonleiomyoma ) . P18887 , hOGG1 , and P21728 were not correlated with the presence of leiomyomas . P18887 (*)GG/GA/AA , hOGG1(*)TT/TA/AA , and P21728 (*)GG/GA/AA were 54.2/37.5/8.3 % , 36.7/44.2/19.1 % , and 3.3/25.8/70.8 % ( leiomyoma ) ; and 48.2/47.3/4.5 % , 43.6/41/15.4 % , and 3.6/25/71.4 % ( nonleiomyoma ) . CONCLUSION(S) : The P38936 codon 31(*)C- and P14416 codon 313(*)T-related genotypes/alleles were associated with the presence of leiomyoma . P18887 , hOGG1 , and P21728 were not correlated with leiomyoma development . Rottlerin , a PKC isozyme-selective inhibitor , affects signaling events and cytokine production in human monocytes . The implication of select protein kinase C ( PKC ) isoenzymes in cytokine production by human monocytes was investigated using an isozyme-selective inhibitor of PKC , rottlerin . We found that lipopolysaccharide ( LPS ) triggers cytosol-to-membrane translocation of PKCalpha and delta isoenzymes , whereas phorbol ester ( PMA ) induces translocation of several PKC isoforms . Moreover , we show that in LPS- and PMA-stimulated monocytes rottlerin affects several cellular responses . ( 1 ) At low ( 15 microM ) concentration it blocks translocation of PKCdelta , diminishes DNA binding activity of AP-1 transcription factor , and attenuates cytokine production [ tumor necrosis factor alpha ( P01375 ) > interleukin-1beta ( IL-1beta ) ] . ( 2 ) At high ( 50 microM ) concentration it prevents translocation of PKCalpha , and subsequently inhibits P27361 / P28482 phosphorylation , DNA binding activities of AP-1 and nuclear factor-KB transcription factors , and the production of both tested cytokines . Thus , we propose that cytosol-to-membrane translocation of PKCalpha and PKdelta isoenzymes may represent early steps in the signaling cascades that lead to P01375 and IL-1beta production in human monocytes . Evidence of an Epigenetic Modification in Cell-cycle Arrest Caused by the Use of Ultra-highly-diluted Gonolobus Condurango Extract . OBJECTIVES : Whether the ultra-highly-diluted remedies used in homeopathy can effectively bring about modulations of gene expressions through acetylation/deacetylation of histones has not been explored . Therefore , in this study , we pointedly checked if the homeopathically-diluted anti-cancer remedy Condurango 30C ( ethanolic extract of Gonolobus condurango diluted 10(-60) times ) was capable of arresting the cell cycles in cervical cancer cells HeLa by triggering an epigenetic modification through modulation of the activity of the key enzyme histone deacetylase 2 vis-a-vis the succussed alcohol ( placebo ) control . METHODS : We checked the activity of different signal proteins ( like P38936 (WAF) , p53 , Akt , P40763 ) related to deacetylation , cell growth and differentiation by western blotting and analyzed cell-cycle arrest , if any , by fluorescence activated cell sorting . After viability assays had been performed with Condurango 30C and with a placebo , the activities of histone de-acetylase ( HDAC ) enzymes 1 and 2 were measured colorimetrically . RESULTS : While Condurango 30C induced cytotoxicity in HeLa cells in vitro and reduced Q92769 activity quite strikingly , it apparently did not alter the Q13547 enzyme ; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity . Data on P38936 (WAF) , p53 , Akt , and P40763 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and P55008 -phase cell-cycle arrest when Condurango 30C was used at a 2 % dose . CONCLUSION : Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells , which the placebo was unable to do . Resistance to P50591 is mediated by Q9UD71 in gastric cancer . PURPOSE : Dopamine and DB02527 -regulated phosphoprotein , Mr 32,000 ( Q9UD71 ) , is overexpressed during the gastric carcinogenesis cascade . Here , we investigated the role of Q9UD71 in promoting resistance to treatment with P50591 . EXPERIMENTAL DESIGN : In vitro cell models including stable expression and knockdown of Q9UD71 were used . The role of Q9UD71 in regulating P50591 -dependent apoptosis was evaluated by clonogenic survival assay , P08758 staining , immunofluorescence , quantitative reverse transcriptase PCR , Western blot , and luciferase reporter assays . RESULTS : Stable expression of Q9UD71 in MKN-28 cells enhanced cell survival and suppressed P50591 -induced cytochrome c release and activation of caspase-8 , -9 , and -3 . Conversely , short hairpin RNA-mediated knockdown of endogenous Q9UD71 sensitized the resistant MKN-45 cells to P50591 -induced apoptosis and enhanced P50591 -mediated activation of caspase-8 , -9 , and -3 . Q9UD71 induced BCL-xL expression through activation of Src/ P40763 signaling , and treatment with the Src-specific inhibitor P50391 abrogated Q9UD71 -dependent BCL-xL upregulation and cell survival in MKN-28 cells . The P50591 treatment induced caspase-dependent cleavage of NF-κBp65 protein ; this cleavage was prevented by Q9UD71 , thus maintaining NF-κB activity and the expression of its target , FLIP(S) protein . This suggests that upregulation of BCL-xL could play a possible role in blocking the mitochondria intrinsic apoptosis pathway , whereas the Q9UD71 effect on the NF-κB/FLIP(S) axis could serve as an additional negative feedback loop that blocks P50591 -induced activation of caspase-8 . CONCLUSION : Our findings uncover a novel mechanism of P50591 resistance mediated by Q9UD71 , whereby it inhibits the intrinsic apoptosis pathway through upregulation of BCL-xL , and the extrinsic apoptosis pathway through the NF-κB/FLIP(S) axis . Integration of porcine chromosome 13 maps . In order to expand the comparative map between human chromosome 3 ( Q9NZP2 ) and porcine chromosome 13 ( SSC13 ) , seven genes from Q9NZP2 were mapped on SSC13 by fluorescence in situ hybridisation ( Q5TCZ1 ) , viz . P09110 , P15309 , O60513 , P02788 , Q15746 , P11177 and P10826 . With a view to integrating this expanded comparative map with the existing SSC13 linkage map , we used the INRA-University of Minnesota porcine Radiation Hybrid panel ( IMpRH ) to localize more precisely and to order 15 genes on the SSC13 map , viz . P15309 , O95622 , P05090 , P06276 , P42081 , P35462 , P17677 , P05166 , P04049 , P08100 , SI , TF , P02786 , Q02880 and Q9UQR1 . In this way , we were able to create an integrated map , containing 38 type I and 81 type II markers , by correlating the linkage , radiation hybrid ( RH ) and cytogenetic maps of SSC13 . This integrated map will give us the opportunity to take maximal advantage of the comparative mapping strategy for positional candidate cloning of genes responsible for economically important traits . Sorting nexin 5 and dopamine d1 receptor regulate the expression of the insulin receptor in human renal proximal tubule cells . Sorting nexin 5 ( Q9Y5X3 ) belongs to the P20073 family , which is composed of a diverse group of proteins that mediate trafficking of plasma membrane proteins , receptors , and transporters . Q9Y5X3 is important in the resensitization of the dopamine D1-like receptor ( D1R ) . D1R is uncoupled from its effector proteins in hypertension and diabetes , and treatment of diabetes restores D1R function and insulin receptor ( IR ) expression . We tested the hypothesis that the D1R and Q9Y5X3 regulate IR by studying the expression , distribution , dynamics , and functional consequences of their interaction in human renal proximal tubule cells ( hRPTCs ) . D1R , Q9Y5X3 , and IR were expressed and colocalized in the brush border of RPTs . P01308 promoted the colocalization of Q9Y5X3 and IR at the perinuclear area of hRPTCs . Unlike Q9Y5X3 , the D1R colocalized and coimmunoprecipitated with IR , and this interaction was enhanced by insulin . To evaluate the role of Q9Y5X3 and D1R on IR signaling , we silenced via RNA interference the endogenous expression of Q9Y5X3 or the D1R gene P21728 in hRPTCs . We observed a decrease in IR expression and abundance of phosphorylated IR substrate and phosphorylated protein kinase B , which are crucial components of the IR signal transduction pathway . Our data indicate that Q9Y5X3 and D1R are necessary for normal IR expression and activity . It is conceivable that D1R and Q9Y5X3 may interact to increase the sensitivity to insulin via a positive regulation of IR and insulin signaling . Interaction of early environment , gender and genes of monoamine neurotransmission in the aetiology of depression in a large population-based Finnish birth cohort . Objectives Depression is a worldwide leading cause of morbidity and disability . Genetic studies have recently begun to elucidate its molecular aetiology . The authors investigated candidate genes of monoamine neurotransmission and early environmental risk factors for depressiveness in the genetically isolated population-based Northern Finland Birth Cohort 1966 ( 12 058 live births ) . Design The authors ascertained and subdivided the study sample ( n=5225 ) based on measures of early development and of social environment , and examined candidate genes of monoamine neurotransmission , many of which have shown prior evidence of a gene-environment interaction for affective disorders , namely P31645 , Q8IWU9 , P21964 , P21397 and the dopamine receptor genes P21728 - P21918 . Results and conclusion The authors observed no major genetic effects of the analysed variants on depressiveness . However , when measures of early development and of social environment were considered , some evidence of interaction was observed . Allelic variants of P21964 interacted with high early developmental risk ( p=0.005 for rs2239393 and p=0.02 for rs4680 ) so that the association with depression was detected only in individuals at high developmental risk group ( p=0.0046 and β=0.056 for rs5993883-rs2239393-rs4680 risk haplotype CGG including Val158 ) , particularly in males ( p=0.0053 and β=0.083 for the haplotype CGG ) . Rs4274224 from P14416 interacted with gender ( p=0.017 ) showing a significant association with depressiveness in males ( p=0.0006 and β=0.0023 ; p=0.00005 and β=0.069 for rs4648318-rs4274224 haplotype GG ) . The results support the role of genes of monoamine neurotransmission in the aetiology of depression conditional on environmental risk and sex , but not direct major effects of monoaminergic genes in this unselected population . Dopamine selectively reduces GABA(B) transmission onto dopaminergic neurones by an unconventional presynaptic action . The functioning of midbrain dopaminergic neurones is closely involved in mental processes and movement . In particular the modulation of the inhibitory inputs on these cells might be crucial in controlling firing activity and dopamine ( DA ) release in the brain . Here , we report a concentration-dependent depressant action of dopamine on the GABA(B) IPSPs intracellularly recorded from dopaminergic neurones . Such effect was observed in spite of the presence of D(1)/ P14416 antagonists . A reduction of the GABA(B) IPSPs was also caused by noradrenaline ( norepinephrine ) and by L-beta-3,4-dihydroxyphenylalanine ( DB01235 ) , which is metabolically transformed into DA . The DA-induced depression of the IPSPs was partially antagonised by the alpha2 antagonists yohimbine and phentolamine . DA did not change the postsynaptic effects of the GABA(B) agonist baclofen , suggesting a presynaptic site of action . Furthermore , DA did not modulate the GABA(A)-mediated IPSP . The DA-induced depression of the GABA(B) IPSP occluded the depression produced by serotonin and was not antagonized by serotonin antagonists . The DA- and 5-HT-induced depression of the GABA(B) IPSP persisted when calcium and potassium currents were reduced in to the presynaptic terminals . These results describe an unconventional presynaptic , D(1) and D(2) independent action of DA on the GABA(B) IPSP . This might have a principal role in determining therapeutic/side effects of DB01235 and antipsychotics and could be also involved in drug abuse . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . A common variant in P35462 gene is associated with risperidone-induced extrapyramidal symptoms . We present a pharmacogenetic study of acute antipsychotic ( AP ) -induced extrapyramidal symptoms ( EPS ) using an extensive linkage disequilibrium mapping approach in seven-candidate genes with a well-established link to dopamine ( P14416 , P35462 , P12821 , P21964 , Q01959 , P21397 , P27338 ) . From a cohort of 321 psychiatric inpatients , 81 cases presenting with EPS ( Simpson-Angus > 3 ) and 189 controls presenting without EPS ( Simpson-Angus < or = 3 ) took part . Eighty-four-tag single nucleotide polymorphisms ( SNPs ) in candidate genes were genotyped . After extensive data cleaning , 70 SNPs were analyzed for association of single markers and haplotypes . AP dosage , AP- P14416 blockade potency and age were identified as susceptibility factors for AP-induced EPS . One SNP of the P35462 gene , rs167771 , achieved significant association with EPS risk after Bonferroni correction ( nominal P-value 1.3 x 10(-4) ) in the patients treated with risperidone ( 132 patients ) . AP-induced EPS remains a serious public health problem . Our finding of a common SNP ( rs167771 ) in the P35462 gene provides a strong new candidate gene for risperidone-induced EPS . Integrative gene network analysis provides novel regulatory relationships , genetic contributions and susceptible targets in autism spectrum disorders . Autism spectrum disorders ( ASDs ) are a group of diseases exhibiting impairment in social drive , communication/language skills and stereotyped behaviors . Though an increased number of candidate genes and molecular interactions have been identified by various approaches , the pathogenesis remains elusive . Based on clinical observations , data from accessible GWAS and expression datasets we identified ASDs gene candidates . Integrative gene network and a novel CNV-centric Node Network ( CNN ) analysis method highlighted ASDs-associated key elements and biological processes . Functional analysis identified neurological functions including synaptic cholinergic receptor ( CHRNA ) families , dopamine receptor ( P14416 ) , and correlations between social behavior and oxytocin related pathways . CNN analysis of genome-wide genetic and expression data identified inheritance-related clusters related to P60484 / Q92574 / Q06787 and mTor/PI3K regulation . Integrative analysis identified potential regulators of networks , specifically P01375 and beta-estradiol , suggesting a potential central role in ASDs . Our data provide information on potential disease mechanisms , and key regulators that may generate novel postulations , and diagnostic molecular biomarkers .

MH_train_53

MH_train_53

MH_train_53

interacts_with DB06699?

multiple_choice

DB06699 : a novel gonadotropin-releasing hormone blocker for the treatment of prostate cancer . Androgen deprivation therapy with gonadotropin releasing-hormone ( DB00644 ) receptor agonists provides the mainstay of endocrine treatment for advanced prostate cancer . Although effective , DB00644 agonists induce an initial testosterone surge , which can cause painful and potentially dangerous clinical flare . DB06699 is a novel P30968 blocker that provides immediate , profound and sustained testosterone reduction , without an initial surge . In a Phase III trial , degarelix and leuprolide showed similar long-term efficacy in maintaining testosterone levels of 0.5 ng/ml or less over 1 year , and induced significantly faster testosterone and prostate-specific antigen suppression . DB06699 was well tolerated ; the most common side effects were mild/moderate injection-site reactions and hot flashes . Findings to date suggest that degarelix may make an important contribution to the treatment of prostate cancer . Modulation of the P22301 /IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis . IFN-beta has been shown to be effective in the treatment of multiple sclerosis ( MS ) . However , the primary mechanism by which IFN-beta mediates its therapeutic effect remains unclear . Recent studies indicate that under defined conditions , IFN-beta may downregulate DC expression of IL-12 . We and others have shown that IFN-beta may also downregulate P22301 . In light of the recently proposed paradigm that an P22301 /IL-12 immunoregulatory circuit controls susceptibility to autoimmune disease , we examined the effect of IFN-beta on the development and behavior of the autoreactive T cell repertoire during experimental autoimmune encephalomyelitis ( EAE ) , an animal model sharing many features with MS . SWXJ mice were immunized with the immunodominant p139-151 determinant of myelin proteolipid protein ( PLP ) , and at onset of EAE were treated every other day with IFN-beta . After eight weeks of treatment , we assessed autoreactivity and observed no significant IFN-beta effect on splenocyte proliferation or splenocyte production of P01579 , P60568 , P05112 , or P05113 in response to the priming determinant used to initiate disease . However , in IFN-beta treated mice , the cytokine profile in response to the priming immunogen was significantly skewed toward an increased production of P22301 and a concurrent decreased production of IL-12 . Moreover , the in vivo modulation of the P22301 /IL-12 immunoregulatory circuit in response to the priming immunogen was accompanied by an aborted development of epitope spreading . Our results indicate that IFN-beta induces a reciprocal modulation of the P22301 /IL-12 cytokine circuit in vivo . This skewed autoreactivity establishes an inflammatory microenvironment that effectively prevents endogenous self-priming thereby inhibiting the progression of disease associated with epitope spreading . Expression of type I P01148 receptor and in vivo and in vitro P01148 -I effects in corpora lutea of pseudopregnant rabbits . The expression of type I P01148 receptor ( P30968 -I ) and the direct role of P01148 -I on corpora lutea ( CL ) function were studied in the pseudopregnant rabbit model . Immunohistochemistry evidenced P30968 -I and P01148 -I in luteal cells at early ( day 4 pseudopregnancy ) - , mid ( day 9 ) - , and late ( day 13 ) -luteal stages . Real-time RT-PCR and western blotting revealed P30968 -I mRNA and protein at the three luteal stages . DB06719 in vivo treatment at days 9 and 13 decreased plasma progesterone levels for 48 and 24 h respectively . In in vitro cultured CL , buserelin reduced progesterone secretion , increased prostaglandin F(2α) ( P49763 (2α) ) secretion and cyclo-oxygenase-2 ( P35354 ) and nitric oxide synthase ( NOS ) activities at days 9 and 13 , and decreased PGE₂ at day 13 . Co-incubation with antagonists for P01148 -I ( antide ) , inositol 1,4,5-trisphosphate ( IP₃ , 2-amino-ethoxydiphenylborate ) , and diacylglycerol ( DAG , 1-hexadecyl-2-acetyl glycerol ) or inhibitors for phospholipase C ( P98160 , compound 48/80 ) , and protein kinase C ( PKC , staurosporine ) counteracted the buserelin effects . DB06719 co-incubated with P36551 inhibitor ( acetylsalicylic acid ) increased progesterone and decreased P49763 (2α) and NOS activity at days 9 and 13 , whereas co-incubation with NOS inhibitor ( DB04223 methyl ester ) increased progesterone at the same luteal stages . These results suggest that P30968 -I is constitutively expressed in rabbit CL independently of luteal stage , whereas P01148 -I down-regulates directly CL progesterone production via P49763 (2α) at mid- and late-luteal stages of pseudopregnancy , utilizing its cognate type I receptor with a post-receptorial mechanism that involves P98160 , IP₃ , DAG , PKC , P35354 , and NOS . Ontogeny of gene expression in the gonadotroph of the developing female rat . During the infantile period of the female rat ( 8-21 postnatal days [ P01160 ] of age ) , there is a dramatic increase in plasma DB00094 , which is thought to be important in initiating ovarian activity and , perhaps , the onset of puberty . To begin to understand the regulation of this DB00094 surge , we determined the ontogenetic development of LHbeta , FSHbeta , and P30968 ( P30968 ) mRNA levels in the pituitary gland throughout the infantile period of the female rat . Steady-state mRNA levels were determined by an external standard quantitative reverse transcriptase polymerase chain reaction assay . FSHbeta and P30968 mRNA levels increased to peak on P01160 12 ( p < 0.03 ) . LHbeta mRNA levels remained relatively constant until rising on P01160 18 . A DB00644 antagonist ( 10-100 microg/animal ) was administered daily from P01160 8-11 or P01160 11-13 , and animals were killed on P01160 12 or P01160 14 , respectively . FSHbeta , LHbeta , and P30968 mRNAs were not affected by DB00644 antagonist treatment . Plasma DB00094 was selectively reduced in the first group , whereas both plasma LH and DB00094 were suppressed in the second group . These data indicate that gene expression of LHbeta , FSHbeta , and P30968 are differentially regulated in the infantile female rat pituitary . DB00644 is involved in regulating the secretion of DB00094 and LH during the infantile period but not in regulating FSHbeta , LHbeta , or P30968 mRNA gene expression . DB06699 . DB06699 is a gonadotropin-releasing hormone ( DB00644 ) receptor antagonist that , in common with P30968 agonists ( e.g. leuprolide , goserelin and triptorelin ) , is indicated for use as an androgen-deprivation therapy in patients with advanced prostate cancer . In 1-year , randomized , open-label , phase II or III trials in patients with all stages of prostate cancer , subcutaneous degarelix was associated with rapid , profound and sustained suppression of serum testosterone and prostate-specific antigen ( PSA ) , without evidence of testosterone surges or microsurges . In the phase III trial , degarelix ( 240 mg initially followed by 80 mg every 28 days ) was considered to be effective and noninferior to intramuscular leuprolide ( 7.5 mg every 28 days ) with regard to inducing and maintaining suppression of serum testosterone to castrate levels ( i.e. < or=0.5 ng/mL ) . DB06699 induced testosterone suppression more rapidly than leuprolide . Median serum testosterone levels of < or=0.5 ng/mL were achieved by day 3 in degarelix recipients , but not until day 28 in leuprolide recipients . PSA suppression was also more rapid with degarelix than with leuprolide , with significant between-group differences in serum PSA levels favouring degarelix at 14 and 28 days . DB06699 treatment for 1 year was generally well tolerated ; the adverse events reported were mostly related to subcutaneous drug administration ( i.e. injection-site reactions ) and hormonal androgen deprivation ( e.g. hot flushes ) . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Large-scale association study for structural soundness and leg locomotion traits in the pig . BACKGROUND : Identification and culling of replacement gilts with poor skeletal conformation and feet and leg ( FL ) unsoundness is an approach used to reduce sow culling and mortality rates in breeding stock . Few candidate genes related to soundness traits have been identified in the pig . METHODS : In this study , 2066 commercial females were scored for 17 traits describing body conformation and FL structure , and were used for association analyses . Genotyping of 121 SNPs derived from 95 genes was implemented using Sequenom 's MassARRAY system . RESULTS : Based on the association results from single trait and principal components using mixed linear model analyses and false discovery rate testing , it was observed that P02649 , P34820 , P30988 , P08123 , P20849 , DKFZ , P35555 and VDBP were very highly significantly ( P < 0.001 ) associated with body conformation traits . The genes P09917 , P34820 , P30988 , O00300 , P30559 and Q9UBV4 were very highly significantly ( P < 0.001 ) associated with FL structures , and P02649 , P30988 , P08123 , P30968 , Q14623 , P42898 and Q9UBV4 were highly significantly ( P < 0.01 ) associated with overall leg action . Strong linkage disequilibrium between P30988 and P08123 on SSC9 was detected , and haplotype -ACGACC- was highly significantly ( P < 0.01 ) associated with overall leg action and several important FL soundness traits . CONCLUSION : The present findings provide a comprehensive list of candidate genes for further use in fine mapping and biological functional analyses . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . Synthesis and biological activity of DB00644 antagonists modified at position 3 with 3-(2-methoxy-5-pyridyl)-alanine . DB06699 is a potent very long-acting DB00644 antagonist after subcutaneous administration . In this paper , we describe the synthesis of two analogs of degarelix incorporating racemic 3-(2-methoxy-5-pyridyl)-alanine ( 2-OMe-5Pal , 5 ) at position 3 . The two diastereomers were separated by reverse-phase high-performance liquid chromatography ( RP-HPLC ) and the absolute stereochemistry at position 3 in the peptides was determined by enzymatic digestion with proteinase K . These analogs were tested in vitro for their ability to antagonize the P30968 and in vivo for duration of action in a castrated male rat assay . Analog 7 with D2-OMe-5Pal was potent in vitro ( IC50 = 5.22 nM ) ; however , analog 8 with Q401N2 -OMe-5Pal at position 3 in degarelix lost potency as an antagonist of the human P30968 ( IC50 = 36.95 nM ) . Both the analogs were found to be short-acting in vivo . Microarray Analysis of the Major Depressive Disorder mRNA Profile Data . OBJECTIVE : Major depressive disorder ( MDD ) is a common mood disorder associated with several psychophysiological changes like disturbances of sleep , appetite , or sexual desire , and it affects the patients ' life seriously . We aimed to explore a genetic method to investigate the mechanism of MDD . METHODS : The mRNA expression profile ( GSE53987 ) of MDD was downloaded from Gene Expression Omnibus database , including 105 samples of three brain regions in post-mortem tissue suffered from MDD and unaffected controls . Differentially expressed genes ( DEGs ) in MDD were identified using the Limma package in R . Gene Ontology functions and Kyoto Enrichment of Genes and Genomes pathways of the selected DEGs were enriched using Database for Annotation , Visualization and Integrated Discovery . Protein-protein interactive network of DEGs was constructed using the Cytoscape software . RESULTS : Totally , 241 DEGs in MDD-hip group , 218 DEGs in MDD-pfc group , and 327 DEGs in MDD-str group were identified . Also , different kinds of biological processes of DEGs in each group were enriched . Besides , glycan biosynthesis of DEGs in MDD-str group , O95786 -like receptor signaling and pyrimidine metabolism of DEGs in the MDD-hip group were enriched , respectively . Moreover , several DEGs like Q05397 , Q13569 and P41208 in MDD-str group , P40126 , AR and P30968 in MDD-pfc group , and P31749 and P51617 in MDD-hip group were selected from PPI network . CONCLUSION : Our data suggests that the brain striatum tissue may be greatly affected by MDD , and DEGs like Q05397 , Q10471 and Q10471 in striatum , AR in prefrontal cortex and P51617 and P29459 in hippocampus may provide novel therapeutic basis for MDD treatment . 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 . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . 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 . Reduced folate carrier and dihydrofolate reductase expression in acute lymphocytic leukemia may predict outcome : a Children 's Cancer Group Study . PURPOSE : DB00563 is a major component of current treatment regimens for children with acute lymphocytic leukemia ( ALL ) . Potential mechanisms of methotrexate resistance include impaired drug uptake , decreased drug retention , and dihydrofolate reductase ( P00374 ) amplification . The purpose of this study was to assess whether reduced folate carrier ( P41440 ) and P00374 expression in untreated leukemic blasts correlated with outcome . METHODS : Quantitative real-time RT-PCR was used to measure P41440 and P00374 mRNA expression in leukemic blasts from 40 newly diagnosed patients with ALL obtained in a blinded fashion from Children 's Cancer Group studies . RESULTS : Low P41440 expression at diagnosis correlated significantly with an unfavorable event free survival . Surprisingly , low , not high , P00374 expression correlated significantly with an unfavorable event-free survival . Proliferative cell nuclear antigen ( P12004 ) expression demonstrated a weak inverse relationship between sample P12004 and P00374 or P41440 expression , suggesting that P00374 and P41440 expression may be markers for factors other than drug resistance . CONCLUSIONS : These results suggest that impaired transport may be an important mechanism of intrinsic methotrexate resistance in ALL , and P00374 expression also may be an important prognostic factor in ALL . Additional studies are necessary to clarify the mechanism for the correlation of low P00374 expression with poor outcome . Expression of gonadotropin-releasing hormone receptor and effect of gonadotropin-releasing hormone analogue on proliferation of cultured gastric smooth muscle cells of rats . AIM : To investigate the expression of gonadotropin-releasing hormone ( DB00644 ) receptor and the effects of DB00644 analog ( alarelin ) on proliferation of cultured gastric smooth muscle cells ( GSMC ) of rats . METHODS : Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of P30968 in GSMC , respectively . Techniques of cell culture , OD value of MTT test , measure of (3)H-TdR incorporation , average fluorescent values of proliferating cell nuclear antigen ( P12004 ) and flow cytometric DNA analysis were used in the experiment . RESULTS : The cultured GSMC of rats showed immunoreactivity for P30968 ; positive staining was located in cytoplasm . P30968 mRNA hybridized signals were also detected in cytoplasm . When alarelin ( 10(-9) , 10(-7) , 10(-5) mol/L ) was administered into the medium and incubated for 24 h , OD value of MTT , (3)H-TdR incorporation and average fluorescent values of P12004 all decreased significantly as compared with the control group ( P < 0.05 ) . The maximum inhibitory effect on cell proliferation was achieved a concentration of 10(-5) mol/L and it acted in a dose-dependent manner . Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G(1) phase and decrease ratio of S phase of GSMC of rats ( P < 0.05 ) . The maximum inhibitory effect on ratio of S phase was at the concentration of 10(-5) mol/L and also acted in a dose-dependent manner . CONCLUSION : Our data suggest that P30968 can be expressed by GSMC of rats . DB00644 analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through DB00644 receptors . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . DB00644 analogues reduce the proliferation of endometrial stromal cells but not endometriotic cells . AIMS : We investigated the potential of gonadotropin-releasing hormone ( DB00644 ) agonists and DB00644 antagonists to inhibit cell proliferation in endometriotic and endometrial stromal cells . METHODS : Twenty patients with ovarian endometriomas and 18 patients with uterine fibromas were recruited . Endometriotic and endometrial stromal cells were obtained from the ovarian chocolate cyst linings and the eutopic endometria of premenopausal women with uterine fibromas , respectively . RESULTS : DB00644 agonist or antagonist treatment attenuated tumor necrosis factor ( P01375 ) -α-induced cell proliferation in the endometrial stromal cells , whereas endometriotic stromal cells did not respond to treatment . The endometriotic stromal cells exhibited a decreased expression of the type I P30968 compared with the endometrial stromal cells . DB00644 agonists or antagonists did not repress P01375 -α-induced P10145 production in endometriotic stromal cells . CONCLUSION : DB00644 agonists and antagonists have similar effects in slowing the growth of endometrial stromal cells . Endometriotic stromal cells resist the antiproliferative effect of DB00644 agonists and antagonists . Serotonin and dopamine receptor gene polymorphisms and the risk of extrapyramidal side effects in perphenazine-treated schizophrenic patients . RATIONALE : DB00850 , a classical antipsychotic drug , has the potential to induce extrapyramidal side effects ( EPS ) . Dopaminergic and serotonergic pathways are involved in the therapeutic and adverse effects of the drug . OBJECTIVES : To evaluate the impact of polymorphisms in the dopamine D(2) and D(3) and serotonin 2A and 2C receptor genes ( P14416 , P35462 , P28223 , and P28335 ) on short-term effects of perphenazine monotherapy in schizophrenic patients . MATERIALS AND METHODS : Forty-seven Estonian inpatients were evaluated before and after 4-6 weeks of treatment by Simpson-Angus rating scale , Barnes scale , and Positive and Negative Symptom Scale . Genotyping was performed for common P14416 , P35462 , P28223 , and P28335 gene polymorphisms , previously reported to influence receptor expression and/or function . RESULTS : Most of the patients ( n = 37 ) responded to the treatment and no significant association was observed between the polymorphisms and antipsychotic response . The 102C allele of P28223 and the -697C and 23Ser alleles of P28335 were more frequent among patients with EPS ( n = 25 ) compared to patients without EPS ( n = 22 ) ( p = 0.02 , 0.01 , and 0.02 , respectively ) . The difference between patients with and without EPS in variant allele frequencies remained significant after multiple model analyses including age , gender , and duration of antipsychotic treatment as covariants . There was no significant association between EPS occurrence and polymorphisms in the P14416 and P35462 genes . CONCLUSIONS : An association was observed between polymorphisms in P28223 and P28335 genes and occurrence of acute EPS in schizophrenic patients treated with perphenazine monotherapy . Larger study populations are needed to confirm our findings . Molecular evolution of the oxytocin-oxytocin receptor system in eutherians . DB00107 ( P01178 ) is a nine-amino-acid peptide hormone that is mainly released at the times of uterine contractions during parturition and milk ejection during lactation , whereas a similar peptide hormone , arginine vasopressin , primarily exerts direct antidiuretic action on the kidney and causes vasoconstriction of the peripheral vessels . The genes coding for these peptides are tandemly located on the same chromosome . A tandem duplication occurring in the common ancestor of jawed vertebrates has been proposed as responsible . In contrast to the two peptide hormones , only one oxytocin receptor ( P30559 ) but three arginine vasopressin receptors ( P37288 , P47901 , and P30518 ) are known ; these receptors probably arose from two rounds of genome duplication in the common ancestor of vertebrates . In this study , we addressed the molecular evolution of the P01178 - P30559 system in eutherians . Our analyses suggest that an amino acid change from isoleucine to lysine on the eighth site ( I8L ) of the peptide , which corresponded to a change from mesotocin to P01178 , had occurred during the common ancestral lineage of eutherians . At around the same time that the emergence of P01178 occurred , functional constraints on the P01178 receptor ( pre- P30559 ) might have relaxed , and a series of nonsynonymous substitutions might have accumulated . Only a few of these nonsynonymous substitutions might have contributed to reestablishing the molecular relationship between the P01178 ligand and its receptor , after which functional constraints on the P30559 were reinstated . Since the P01178 - P30559 system plays an important role in eutherians , the evolution of the P01178 - P30559 system was probably an essential component of the genesis of the eutherian signature . Evaluation of degarelix in the management of prostate cancer . Medical castration using gonadotropin-releasing hormone ( DB00644 ) receptor agonists currently provides the mainstay of androgen deprivation therapy for prostate cancer . Although effective , these agents only reduce testosterone levels after a delay of 14 to 21 days ; they also cause an initial surge in testosterone that can stimulate the cancer and lead to exacerbation of symptoms ( " clinical flare " ) in patients with advanced disease . Phase III trial data for the recently approved P30968 blocker , degarelix , demonstrated that it is as effective and well tolerated as DB00644 agonists . However , it has a pharmacological profile more closely matching orchiectomy , with an immediate onset of action and faster testosterone and PSA suppression , without a testosterone surge or microsurges following repeated injections . As a consequence , with this DB00644 blocker , there is no risk of clinical flare and no need for concomitant antiandrogen flare protection . DB06699 therefore provides a useful addition to the hormonal armamentarium for prostate cancer and offers a valuable new treatment option for patients with hormone-sensitive advanced disease . Here , we review key preclinical and clinical data for degarelix , and look at patient-focused perspectives in the management of prostate cancer . Serotonergic mechanisms in human allergic contact dermatitis . Expression of serotonin ( 5-hydroxytryptamine ; 5-HT ) , 5-HT receptors 1A ( 5-HT1AR ) and 2A , and serotonin transporter protein ( P31645 ) was studied in positive epicutaneous reactions to nickel sulphate in nickel-allergic patients , at 72 h post-challenge with the antigen . In addition , the effects of 5-HT2AR agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , and the selective serotonin reuptake inhibitors ( SSRIs ) citalopram and fluoxetine , were tested on nickel-stimulated peripheral blood mononuclear cells from nickel-allergic patients , regarding their proliferation and interleukin ( IL ) -2 production , as well as the effect of these SSRIs on a murine Langerhans ' cell-like line ( XS52 ) , regarding its IL-1beta production . Serotonin-positive platelets were increased in the inflamed skin compared with control skin . A decrease ( p < 0.01 ) in 5-HT1AR-positive mononuclear cells was evident in the eczematous skin compared with control skin , whereas 5-HT2AR- and P31645 -positive cells were increased ( p < 0.001 for both ) in the eczematous skin . Treatment of nickel-stimulated peripheral blood mononuclear cells with 5x10(-5) mol/l of DOI inhibited ( p < 0.01 ) the proliferation of nickel-stimulated peripheral blood mononuclear cells , while no effect was found regarding P60568 production . DB00215 at 10(-6) mol/l tended to inhibit the production of IL-1beta by the XS52 cell line . These results indicate the implication of the serotonergic system in the contact allergic reaction . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . [ The efficacy of degarelix on LUTS ( Lower urinary tract symptoms ) relief in patients with prostate cancer ] . Hormonal therapy is one of the treatment options for prostate cancer patients . There are many hormonal treatments modality to block the testosterone effect on prostate cancer cell proliferation . DB06699 is an innovative molecule able to antagonize the P30968 with comparable oncological results to DB00644 agonist , but with less side effects , avoiding the flare up phase , and better efficacy in LUTS relief . These characteristics of degarelix can impact on the clinical decision making to choose a therapy instead of another . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers .

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P43119 -mediated DB00171 release from erythrocytes requires the voltage-dependent anion channel . Erythrocytes have been implicated as controllers of vascular caliber by virtue of their ability to release the vasodilator DB00171 in response to local physiological and pharmacological stimuli . The regulated release of DB00171 from erythrocytes requires activation of a signaling pathway involving G proteins ( G(i) or G(s) ) , adenylyl cyclase , protein kinase A , and the cystic fibrosis transmembrane conductance regulator as well as a final conduit through which this highly charged anion exits the cell . Although pannexin 1 has been shown to be the final conduit for DB00171 release from human erythrocytes in response to reduced oxygen tension , it does not participate in transport of DB00171 following stimulation of the prostacyclin ( IP ) receptor in these cells , which suggests that an additional protein must be involved . Using antibodies directed against voltage-dependent anion channel ( P21796 )1 , we confirm that this protein is present in human erythrocyte membranes . To address the role of P21796 in DB00171 release , two structurally dissimilar P21796 inhibitors , Bcl-x(L) BH4(4-23) and DB05185 , were used . In response to the IP receptor agonists , iloprost and UT-15C , DB00171 release was inhibited by both P21796 inhibitors although neither iloprost-induced DB02527 accumulation nor total intracellular DB00171 concentration were altered . Together , these findings support the hypothesis that P21796 is the DB00171 conduit in the IP receptor-mediated signaling pathway in human erythrocytes . In addition , neither the pannexin inhibitor carbenoxolone nor Bcl-x(L) BH4(4-23) attenuated DB00171 release in response to incubation of erythrocytes with the β-adrenergic receptor agonist isoproterenol , suggesting the presence of yet another channel for DB00171 release from human erythrocytes . P43119 up-regulates the expression of angiogenic genes in human endometrium via cross talk with epidermal growth factor Receptor and the extracellular signaling receptor kinase 1/2 pathway . DB01240 ( P06744 ) is a member of the prostanoid family of lipid mediators that mediates its effects through a seven-transmembrane G protein-coupled receptor ( IP receptor ) . Recent studies have ascertained a role for prostanoid-receptor signaling in angiogenesis . In this study we examined the temporal-spatial expression of the IP receptor within normal human endometrium and additionally explored the signaling pathways mediating the role of IP receptor in activation of target angiogenic genes . Quantitative RT-PCR analysis demonstrated the highest endometrial expression of the IP receptor during the menstrual phase compared with all other stages of the menstrual cycle . Immunohistochemical analysis localized the site of IP receptor expression to the glandular epithelial compartment with stromal and perivascular cell immunoreactivity . Expression of the immunoreactive IP receptor protein was greatest during the proliferative and early secretory phases of the menstrual cycle . To explore the role of the IP receptor in glandular epithelial cells , we used the Ishikawa endometrial epithelial cell line . Stimulation of Ishikawa cells and human endometrial biopsy explants with 100 nm iloprost ( a P06744 analog ) rapidly activated P27361 /2 signaling and induced the expression of proangiogenic genes , basic fibroblast growth factor , angiopoietin-1 , and angiopoietin-2 , in an epidermal growth factor receptor ( P00533 ) -dependent manner . Furthermore , P00533 colocalized with IP receptor in the glandular epithelial compartment . These data suggest that P06744 -IP interaction within glandular epithelial cells can promote the expression of proangiogenic genes in human endometrium via cross talk with the P00533 . [ Thrombocyte prostacyclin receptors in gestational hypertension and pre-eclampsia ] . OBJECTIVE : To determine prostacyclin ( DB01240 ) receptor characteristics in pregnancies complicated by hypertension and to assess any relation to the clinical outcome . METHODS : Radioligand binding studies with [ 3H ] - DB01088 were performed to measure receptor capacity ( Bmax ) and affinity ( Kd-1 ) using platelet membranes from patients with preeclampsia , gestational hypertension or normal pregnancy . RESULTS : P43119 capacity did not differ between the patient groups . In contrast , P43119 affinity was diminished in gestational hypertension and considerably reduced in preeclampsia compared to normal pregnancy . A similar pattern was found in fetal growth ( normal pregnancy > gestational hypertension > preeclampsia ) . Furthermore , the rate of low Apgar scores and acidosis was increased in preeclampsia . CONCLUSIONS : In preeclampsia reduced platelet P43119 affinity was found as well as poor pregnancy outcome in comparison with normal pregnancy , whereas these differences were less pronounced in gestational hypertension . This suggests a role of DB01240 and its receptor in gestational hypertension and in particular in preeclampsia . Induction of prostacyclin receptor expression in human erythroleukemia cells . We have identified both high-affinity ( KD = 36 +/- 3 nM ) and low-affinity ( KD = 2.1 +/- 0.8 microM ) prostacyclin ( DB01240 ) -receptor sites on human erythroleukemia ( HEL ) cells using the radiolabelled prostacyclin analogue . [3H]iloprost . The addition of the phorbol ester , TPA , to the culture medium caused a 5-10-fold increase in the number of both the low- and the high-affinity sites , without any change in their affinity constants . DB01088 stimulated HEL cell membrane adenylate cyclase activity 5-fold . This stimulation was potentiated in the presence of GTP , indicating a conventional P43119 -G2-adenylate cyclase system . HEL cells represent a source of prostacyclin receptor mRNA which may be of value in expression cloning of this receptor . Human DB05914 provide stromal support for efficient P28906 + transduction . Human DB05914 ( hMSC ) -nonhematopoietic cells within the bone marrow microenvironment that can be culture expanded to a uniform population of fibroblastic cells-have been shown to support long-term hematopoiesis of P28906 + cells . Because direct contact between stromal elements and P28906 + cells enhances long-term engraftment , we postulated that hMSC would be a good alternative to the more heterogeneous stroma currently used in gene transfer studies . We used hMSC to support retroviral gene transfer of the G156A P16455 ( deltaMGMT ) gene encoding an alkyltransferase ( AGT ) , which confers drug resistance to a combination of O6-benzylguanine ( BG ) plus the alkylating agents DB00262 and temozolomide ( DB00853 ) in human hematopoietic progenitors . In the presence of P08700 , P05231 , P21583 , or leukemia inhibitory factor ( P15018 ) and Flt-3 ligand , hMSC facilitated expansion and retroviral transduction of human peripheral blood-mobilized P28906 + cells . Furthermore , the transduced cells expressed AGT in 29 % of hematopoietic cells and were 5-fold more resistant to DB00262 and DB00853 than were untransduced cells . Unirradiated hMSC present as support cells were simultaneously transduced and expressed AGT in 26 % of the cells . Thus , the homogeneous nature of hMSC , and their ability to support gene transfer and be transduced themselves suggest they may be useful in clinical gene transfer protocols and have broad therapeutic applications . [ Human monocyte-derived multinucleated giant cells ] . Multinucleated giant cells ( MGC ) are characteristic cells in granulomatous disorders such as sarcoidosis and leprosy . There are two types of MGC ; foreign body-type and Langhans-type cells . The exact mechanisms of the formation and the functional significance of MGC are not determined , although their morphological features are well understood . MGC are also formed in vitro from peripheral blood mononuclear cells by stimulation with cytokines and lectins . Particularly P01579 is considered to play a pivotal role in monocyte fusion . P08700 , P05112 , P35225 , and GM- P04141 are other reported cytokines involved in MGC formation . In addition to such inflammatory mediators , a factor derived from the pathogens of granulomatous disorders may be necessary for MGC formation . Muramyl dipeptide , a peptidoglycan portion of bacterial cell walls , is one of the candidates and can preferentially induce Langhans-type cells in in vitro MGC formation system . Although the exact mechanisms of in vitro MGC formation remains unknown , cell surface molecules such as Q99572 receptor , integrins , CD98 , and macrophage fusion protein are considered to be involved in fusion process . Monocytes of sarcoidosis patients expressed higher levels of Q99572 and had a higher ability to induce MGC than those of healthy controls . Effective agents for sarcoidosis such as tranilast , alloprinol , and captopril inhibited in vitro MGC formation , suggesting their therapeutic effects through the direct effects on monocytes . Thus , an in vitro MGC formation model would be a useful tool to understand the relevance of MGC in granulomatous disorders . Microsomal transfer protein ( P55157 ) inhibition-a novel approach to the treatment of homozygous hypercholesterolemia . Homozygous familial hypercholesterolemia ( HoFH ) represents the most severe lipoprotein disorder , generally attributable to mutation(s) of the low-density lipoprotein receptor ( LDL-R ) , i.e. autosomal dominant hypercholesterolemia type 1 ( P07327 ) . Much lower percentages are due to alterations of apolipoprotein B ( P00325 ) , or gain-of-function mutations of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) ( P00326 ) . In certain geographical areas a significant number of patients may be affected by an autosomal recessive hypercholesterolemia ( Q5SW96 ) . Mutations may be also combined ( two mutations of the same gene , compound heterozygosity ) , or two in different genes ( double heterozygosity ) . Among the most innovative therapeutic approaches made available recently , inhibitors of the microsomal transfer protein ( P55157 ) system have shown a high clinical potential . P55157 plays a critical role in the assembly/secretion of very-low-density lipoproteins ( VLDL ) , and its absence leads to apo B deficiency . P55157 antagonists dramatically lower LDL-cholesterol ( LDL-C ) in animals , although a reported increase of liver fat delayed their clinical development . DB08827 , the best-studied P55157 inhibitor , reduces LDL-C by 50 % or more in HoFH patients , with modest , reversible , liver steatosis . Recent US approval has confirmed an acceptable tolerability , provided patients adhere to a strictly low-fat regimen . There are no clinical data on atherosclerosis reduction/regression , but animal models provide encouraging results . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . P43119 induces P42224 and P40763 phosphorylations in human erythroleukemia cells : a mechanism requiring PTX-insensitive G proteins , P29323 and JNK . The ability of the human prostacyclin receptor ( hIP ) to regulate the activities of signal transducers and activators of transcription ( STATs ) has not yet been documented . In the present study , we have delineated the mechanism by which hIP induces P40763 phosphorylations in human erythroleukemia ( HEL ) cells . Stimulation of endogenous hIP by its specific agonist , cicaprost , resulted in P40763 Tyr705 and Ser727 phosphorylations in a time- and concentration-dependent manner . Cicaprost-induced P40763 Tyr705 and Ser727 phosphorylations were resistant to pertussis toxin ( PTX ) treatment , suggesting that these responses were mediated through PTX-insensitive G proteins . In addition , extracellular signal-regulated kinase ( P29323 ) and c-Jun N-terminal kinase ( JNK ) , but not p38 MAPK , were shown to be phosphorylated by cicaprost in a time- and concentration-dependent manner via PTX-insensitive G proteins . The levels of the interaction between P40763 , P29323 and JNK were enhanced by cicaprost treatment . The involvement of P04049 , Q02750 /2 and JNK in cicaprost-induced phosphorylations of P40763 was illustrated by the use of their selective inhibitors . In contrast , p38 MAPK did not appear to be required . Similar observations were obtained with P42224 upon stimulation by cicaprost . Taken together , these results demonstrate for the first time that hIP activation by cicaprost can lead to P42224 and P40763 phosphorylations via signaling pathways involving PTX-insensitive G proteins , P29323 and JNK . Catecholamine-producing cells in the synovial tissue during arthritis : modulation of sympathetic neurotransmitters as new therapeutic target . BACKGROUND : The proinflammatory and anti-inflammatory role of the sympathetic nervous system in early and late inflammation is an unresolved paradox . A drastic loss of sympathetic nerve fibres in the synovial tissue of patients with rheumatoid arthritis ( RA ) has previously been demonstrated . The presence of tyrosine hydroxylase ( TH ) -positive cells in RA and osteoarthritis ( OA ) has been determined , but the role of these cells in inflammation is still unclear . OBJECTIVE : To characterise TH-positive cells in inflamed RA and OA synovial tissue and to study their role in inflammation . METHODS : Synovial samples were obtained from 32 patients with OA and 19 patients with RA and from 10 control patients . Synovial tissue samples were used for immunofluorescence staining . Synovial cells were isolated by tissue digestion and immediately used for cell culture . For in vivo experiments , collagen type-II arthritis in DBA/1J mice was induced . RESULTS : TH+ cells were present only in inflamed tissue and not in controls . Catecholamine-storing vesicles and vesicular monoamine transporter 2 ( Q05940 ) were identified in the synovial tissue . Experimental increase of cytoplasmic catecholamines by Q05940 blockade strongly reduced tumour necrosis factor ( P01375 ) independently of canonical extracellular β-adrenergic signalling . In addition , Q05940 blockade increased cyclic AMP ( DB02527 ) and DB02527 responsive element binding protein , responsible for P01375 inhibition . In vivo , appearance of Q05940 positive cells was confirmed . Q05940 blockade ameliorated inflammation also in vivo . CONCLUSIONS : This study demonstrates that local catecholamine-producing cells start to replace sympathetic nerve fibres around the onset of disease , and modulation of locally produced catecholamines has strong anti-inflammatory effects in vivo and in vitro . DB01088 has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 + T cells was measured by ELISA . The expression of the transcription factor FoxP3 after co-culture of DCs with P01730 + CD45RA+ T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 , P05231 , P10145 and IL-12p70 in mDCs , while it enhanced P22301 production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA+ T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP3 expression and P22301 production . Additionally , iloprost inhibited the MIP-3beta-induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans . A protective role of hydrogen sulfide against oxidative stress in rat gastric mucosal epithelium . We investigated effect of hydrogen sulfide ( H(2)S ) on oxidative stress-caused cell death in gastric mucosal epithelial cells . In rat normal gastric epithelial RGM1 cells , NaHS , a H(2)S donor , at 1.5mM strongly suppressed hydrogen peroxide ( H(2)O(2) ) -caused cell death , while it slightly augmented the H(2)O(2) toxicity at 0.5-1mM . The protective effect of NaHS was abolished by inhibitors of MEK or JNK , but not of p38 Q96HU1 kinase . NaHS at 1.5mM actually phosphorylated P29323 and JNK in RGM1 cells . DB01016 , an DB00171 -sensitive K(+) ( K( DB00171 )(+) ) channel inhibitor , did not affect the protective effect of NaHS , although mRNAs for K( DB00171 )(+) channel subunits , Kir6.1 and Q09428 , were detected in RGM1 cells . In anesthetized rats , oral administration of NaHS protected against gastric mucosal lesion caused by ischemia-reperfusion . These results suggest that NaHS/H(2)S may protect gastric mucosal epithelial cells against oxidative stress through stimulation of Q96HU1 kinase pathways , a therapeutic dose range being very narrow . Concise prediction models of anticancer efficacy of 8 drugs using expression data from 12 selected genes . We developed concise , accurate prediction models of the in vitro activity for 8 anticancer drugs ( DB00544 , DB00515 , DB00305 , DOX , CPT-11 , SN-38 , TXL and TXT ) , along with individual clinical responses to DB00544 using expression data of 12 genes . We first performed cDNA microarray analysis and MTT assay of 19 human cancer cell lines to sort out genes which were correlative in expression levels with cytotoxicities of the 8 drugs ; we selected 13 genes with proven functional significance to drug sensitivity from a huge number of potent prediction marker genes . The correlation significance of each was confirmed using expression data quantified by real-time RT-PCR , and finally 12 genes ( P08183 , Q9UNQ0 , P10632 , P08684 , Q12882 , P09211 , P16455 , P15559 , P16435 , P11388 , P07437 and P04818 ) were selected as more reliable predictors of drug response . Using multiple regression analysis , we fixed 8 prediction formulae which embraced the variable expressions of the 12 genes and arranged them in order , to predict the efficacy of the drugs by referring to the value of Akaike 's information criterion for each sample . These formulae appeared to accurately predict the in vitro efficacy of the drugs . For the first clinical application model , we fixed prediction formulae for individual clinical response to DB00544 in the same way using 41 clinical samples obtained from 30 gastric cancer patients and found to be of predictive value in terms of survival , time to treatment failure and tumor growth . None of the 12 selected genes alone could predict such clinical responses . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Red meat and poultry , cooking practices , genetic susceptibility and risk of prostate cancer : results from a multiethnic case-control study . Red meat , processed and unprocessed , has been considered a potential prostate cancer ( DB11245 ) risk factor ; epidemiological evidence , however , is inconclusive . An association between meat intake and DB11245 may be due to potent chemical carcinogens that are generated when meats are cooked at high temperatures . We investigated the association between red meat and poultry intake and localized and advanced DB11245 taking into account cooking practices and polymorphisms in enzymes that metabolize carcinogens that accumulate in cooked meats . We analyzed data for 1096 controls , 717 localized and 1140 advanced cases from the California Collaborative Prostate Cancer Study , a multiethnic , population-based case-control study . We examined nutrient density-adjusted intake of red meat and poultry and tested for effect modification by 12 SNPs and 2 copy number variants in 10 carcinogen metabolism genes : P09211 , P35354 , P05177 , P05181 , P07099 , Q16678 , P19224 , NAT2 , P09488 and P30711 . We observed a positive association between risk of advanced DB11245 and high intake of red meat cooked at high temperatures ( trend P = 0.026 ) , cooked by pan-frying ( trend P = 0.035 ) , and cooked until well-done ( trend P = 0.013 ) . An inverse association was observed for baked poultry and advanced DB11245 risk ( trend P = 0.023 ) . A gene-by-diet interaction was observed between an SNP in the P35354 gene and the estimated levels of meat mutagens ( interaction P = 0.008 ) . Our results support a role for carcinogens that accumulate in meats cooked at high temperatures as potential DB11245 risk factors , and may support a role for heterocyclic amines ( HCAs ) in DB11245 etiology . Altered expression of inflammation-related genes in human carotid atherosclerotic plaques . OBJECTIVE : Inflammation is a pivotal process in atherosclerosis development and progression , but the underlying molecular mechanisms remain largely obscure . We have conducted an extensive expression study of atherosclerotic plaques to identify the inflammatory pathways involved in atherosclerosis . METHODS : We studied 11 human carotid plaques , their respective adjacent regions and 7 control arteries from different subjects . Expression of 92 genes was studied by TaqMan low-density array human inflammation panel . Human aortic endothelial and smooth muscle cells were used for in vitro experiments . RESULTS : The mRNA levels of 44/92 genes ( 48 % ) differed significantly between the tissues examined ( 13 up-regulated and 31 down-regulated ) . Dysregulated genes encode molecules belonging to different functional classes although most of them encode enzymes involved in the eicosanoid synthesis pathway . The expression of Q16647 and P43119 genes was decreased in human aortic endothelial and smooth muscle cells stimulated with oxLDL and P01375 -α . CONCLUSIONS : This study not only reveals several dysregulated genes in human lesions but also focuses the role played by the genes involved in the eicosanoid synthesis pathway during atherosclerotic development . The decrease of Q16647 and P43119 expression after oxLDL treatment mirrors the decreased mRNA levels in atherosclerotic lesions versus control arteries , which suggests that oxidation is important for Q16647 and P43119 regulation in human vessel cells during atherosclerosis development . Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 -induced survival . This inhibition can not be overcome by increasing concentrations of P05113 and is not due to the blocking of Na+ channels by lidocaine . Here we report that one class of K+ channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 . In contrast , increasing concentrations of P08700 or granulocyte-macrophage P04141 overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 -sensitive K+ channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [3H]glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . Inhibitors of DB00171 -binding cassette transporters suppress interleukin-12 p40 production and major histocompatibility complex II up-regulation in macrophages . DB00171 -binding cassette ( DB01048 ) transporters are a large family of proteins whose role is to translocate various substances across biological membranes . They include the Tangier disease protein ABC1 , sulfonylurea receptors ( Q09428 ) , multidrug resistance protein ( MDR ) , and cystic fibrosis transmembrane regulator ( P13569 ) . In the current study , we investigated the involvement of ABC transporters in the regulation of lipopolysaccharide ( LPS ) and/or interferon ( IFN ) -gamma-induced interleukin ( IL ) -12 p40 and tumor necrosis factor ( P01375 ) -alpha production , nitric oxide formation , as well as major histocompatibility complex II up-regulation in macrophages . The general ABC transporter inhibitor glibenclamide suppressed both IL-12 p40 and nitric oxide production . However , glibenclamide failed to affect the production of P01375 . The selective ABC1 inhibitors 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and sulfobromophthalein mimicked the suppressive effect of glibenclamide on IL-12 p40 production . On the other hand , both the MDR inhibitor verapamil and P13569 blocker 2,2'-iminodibenzoic acid failed to suppress the production of IL-12 p40 . Furthermore , selective inhibitors and activators of SURs were without effect . In agreement with the pharmacological data , macrophages expressed mRNA for ABC1 , but not SURs or P13569 . Intracellular levels of IL-12 p40 were decreased by glibenclamide , suggesting that glibenclamide does not affect IL-12 p40 secretion . The effect of glibenclamide did not involve an interference with the activation of the p38 and Q8NFH3 /44 mitogen-activated protein kinases or c-Jun kinase . DB01016 also suppressed P01579 -induced up-regulation of major histocompatibility complex II . Taken together , our results indicate that ABC proteins regulate LPS and/or P01579 -induced macrophage activation . Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5-linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter ---- MLVI-2 ---- cen ---- P07686 ---- P00374 ---- Pi227- --- cp12.6 ---- ( P05113 , P05112 ) ---- P08700 ---- P04141 ---- P05230 ---- ( P07333 , P09619 ) ---- ( treC,ADRBR ) ---- ( Q5SW96 - Q13585 , P09603 ) ---- qter . The suggested order and orientation for the closely linked P08700 / P04141 gene pair is cen ---- 5' P08700 3' ---- 5' P04141 3' ---- qter , on the basis of analysis of the P04141 rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q- analyses , was telomeric to P04141 and centromeric to P07333 / P09619 , near P05230 . Long-range restriction-enzyme analysis of 5q- DNAs did not detect rearrangements of 5q-linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12.6 , P08700 , and P07333 can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions . 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 . The anticoagulant effect of PGI2S and tPA in transgenic umbilical vein endothelial cells is linked to up-regulation of PKA and PKC . The selection of vascular grafts for coronary artery bypass surgery is crucial for a positive outcome . This study aimed to establish a novel line of vascular endothelial cells with a potent anticoagulant effect . A lentiviral vector was used to stably transfect human umbilical vein endothelial cells ( HUVECs ) with PGI2S alone ( HUVEC-PGI2S ) or both PGI2S and tPA ( HUVEC-PGI2S-tPA ) . Both HUVEC-PGI2S and HUVEC-PGI2S-tPA cells over-expressing PGI2S and tPA were compared to mock-transfected cells . The enzyme-linked immuno sorbent assay ( ELISAs ) demonstrated that the anticoagulation components , P01008 and P00747 , were up-regulated and coagulation factor FVIII was down-regulated in both cell lines . QRT-PCR and western blotting demonstrated the vasodilation and platelet disaggregation proteins PKA , PKC , and P43119 were up-regulated in both cell lines , but MAPK expression was not altered in either cell line . However , cell viability and colony formation assays and cell cycle analysis demonstrated that both cell lines had a lower rate of cell growth and induced P55008 phase arrest . HUVEC-PGI2S and HUVEC-PGI2S-tPA cells have a potent anticoagulant effect and their use in vascular heterografts may decrease the risk of thrombosis . Regulation of cyclooxygenase-2 and endogenous cytokine expression by bacterial lipopolysaccharide that acts in synergy with c-kit ligand and Fc epsilon receptor I crosslinking in cultured mast cells . Emerging evidence has suggested the pivotal role of mast cells in a host defense against bacterial infection . In this paper , we report that bacterial lipopolysaccharide ( LPS ) is a potent enhancer of the cytokine- and IgE-dependent delayed responses of P08700 -dependent mouse bone marrow-derived cultured mast cells ( BMMC ) . LPS , although showing minimal effects , significantly augmented the c-kit ligand ( KL ) - or IgE-dependent expression of cyclooxygenase ( P36551 ) -2 and the attendant delayed PGD2 generation , with P22301 and P05112 acting as potentiating and inhibitory cytokines , respectively . The P35354 -inducing activity of LPS was mimicked by exogenous P01584 . Assessment of endogenous cytokine induction revealed that P01584 expression was stimulated by either LPS or exogenous P01584 . P05231 expression occurred in parallel with P35354 expression . P22301 expression , which lagged behind P35354 expression , depended on exogenous P22301 , but not on LPS and P01584 . Thus , LPS and P01584 exhibited similar biological activities in terms of P35354 and endogenous cytokine expression . However , adding an antibody against the type I IL-1 receptor to BMMC , which abrogated the effects of P01584 , failed to neutralize the effects of LPS . These results suggest that LPS activates BMMC through the signal transduction pathway shared with exogenous P01584 , rather than exerting its action indirectly via the production of endogenous P01584 . P35354 -derived prostacyclin confers atheroprotection on female mice . Female gender affords relative protection from cardiovascular disease until the menopause . We report that estrogen acts on estrogen receptor subtype alpha to up-regulate the production of atheroprotective prostacyclin , DB01240 , by activation of cyclooxygenase 2 ( P35354 ) . This mechanism restrained both oxidant stress and platelet activation that contribute to atherogenesis in female mice . Deletion of the P43119 removed the atheroprotective effect of estrogen in ovariectomized female mice . This suggests that chronic treatment of patients with selective inhibitors of P35354 could undermine protection from cardiovascular disease in premenopausal females . The uremic toxin 3-indoxyl sulfate is a potent endogenous agonist for the human aryl hydrocarbon receptor . The aryl hydrocarbon receptor ( P35869 ) is a ligand-activated transcription factor involved in the regulation of multiple cellular pathways , such as xenobiotic metabolism and Th17 cell differentiation . Identification of key physiologically relevant ligands that regulate P35869 function remains to be accomplished . Screening of indole metabolites has identified indoxyl 3-sulfate ( I3S ) as a potent endogenous ligand that selectively activates the human P35869 at nanomolar concentrations in primary human hepatocytes , regulating transcription of multiple genes , including P04798 , P05177 , Q16678 , P22309 , P19224 , P05231 , and P0DJI8 . Furthermore , I3S exhibits an approximately 500-fold greater potency in terms of transcriptional activation of the human P35869 relative to the mouse P35869 in cell lines . Structure-function studies reveal that the sulfate group is an important determinant for efficient P35869 activation . This is the first phase II enzymatic product identified that can significantly activate the P35869 , and ligand competition binding assays indicate that I3S is a direct P35869 ligand . I3S failed to activate either CAR or O75469 . The physiological importance of I3S lies in the fact that it is a key uremic toxin that accumulates to high micromolar concentrations in kidney dialysis patients , but its mechanism of action is unknown . I3S represents the first identified relatively high potency endogenous P35869 ligand that plays a key role in human disease progression . These studies provide evidence that the production of I3S can lead to P35869 activation and altered drug metabolism . Our results also suggest that prolonged activation of the P35869 by I3S may contribute to toxicity observed in kidney dialysis patients and thus represent a possible therapeutic target . What future for combination therapies ? For most patients who require lipid-lowering treatment , statin monotherapy is the appropriate treatment . However , in those patients where statin monotherapy does not produce optimal lipid levels , the combination of a statin with niacin , a bile acid sequestrant , a fibric acid derivative , a cholesterol absorption inhibitor or a fish oil preparation may provide improved control . The choice of combination therapy depends upon the patient 's lipid profile and tolerability of the medication . Combination of a statin with niacin , a bile acid sequestrant or ezetimibe , a cholesterol absorption inhibitor , should be considered for patients with very high low-density lipoprotein cholesterol ( LDL-C ) levels , while combination with either a fibric acid derivative or a fish oil should be considered for patients with high LDL-C and high triglyceride levels . A number of new lipid-lowering agents are currently in development , including cholesteryl ester transfer protein ( P11597 ) inhibitors , acyl coenzyme A : cholesterol acyltransferase ( ACAT ) inhibitors , ileal bile acid transport ( Q12908 ) inhibitors , microsomal triglyceride transfer protein ( P55157 ) inhibitors and dual peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma agonists . Introduction of these novel therapies will provide opportunities for developing different combination strategies that may help to optimise lipid profiles in patients who are currently difficult to treat . The introduction of new combinations will require careful study to ensure that the risks of drug interactions and adverse events are minimised . P01375 polymorphisms as a potential modifier gene in the cystic fibrosis . Modifier genes , as the P01375 -α gene , can modulate the cystic fibrosis ( CF ) severity . Thus , -238G > A and -308G > A polymorphisms of P01375 -α gene were analyzed as modifiers of CF . In this context , the present study enrolled 49 CF patients ( diagnosis performed by sweat test and complete P13569 screening ) . The -238G > A polymorphism analysis was performed by Q9ULH0 -PCR , and -308G > A , by PCR-RFLP . In our data , the -238G > A polymorphism was not associated with clinical variability . The AA genotype for -308G > A polymorphism was a risk factor for early gastrointestinal symptoms ( OR=5.98 , 95 % CI=1.06-49.68 ) and protection for the first Pseudomonas aeruginosa ( OR=0.05 , 95 % CI=0.0003-0.007 ) . For the first P. aeruginosa , GA genotype was a risk factor ( OR=10.2 , 95 % CI=1.86-84.09 ) ; for the same genotype , the diagnosis was made in minor time than the AA genotype ( p=0.031 ) . Considering the -308G > A polymorphism alleles , the G allele was a risk factor for early pulmonary symptoms ( OR=3.81 , 95 % CI=1.13-12.97 ) and P. aeruginosa ( OR=66.77 , 95 % CI=15.18-482.7 ) ; however , the same allele showed better transcutaneous oxygen saturation ( OR=9.24 , 95 % CI=1.53-206.1 ) . The A allele was a protective factor for early pulmonary symptoms ( OR=12.26 , 95 % CI=0.08-0.89 ) and P. aeruginosa ( OR=12.15 , 95 % CI=0002-0007 ) , however , the same allele was a risk factor for worst transcutaneous oxygen saturation ( OR=7.01 , 95 % CI=1.14-157.4 ) . As conclusion , the -308G > A polymorphism of the P01375 -α gene was associated with the CF severity . Developmental regulation of prostacyclin synthase and prostacyclin receptors in the ovine uterus and conceptus during the peri-implantation period . This study documents the expression of prostacyclin ( DB01240 ) synthase ( Q16647 ) and DB01240 receptors in the trophoblast and uterus of the ewe at the time of maternal recognition of pregnancy ( i.e. days 7 , 9 , 12 , 14 and 17 ) . The membrane receptor for DB01240 ( P43119 ) and the nuclear receptors , i.e. peroxisome proliferator-activated receptors ( Q07869 ) and their heterodimer partners the retinoid X receptors ( RXR ) , were analysed . In the endometrium , Q16647 transcript and protein were expressed at day 9 of pregnancy and levels declined from days 12 to 17 . Immunohistochemistry and in situ hybridization indicated that Q16647 was mainly located in the luminal epithelium of the endometrium . Endometrial P43119 , Q07869 , P37231 and P48443 expression was regulated during the peri-implantation period whereas Q03181 , P19793 and P28702 were consistently expressed . In the trophoblast , Q16647 transcript levels rose as development progressed and peaked at day 17 . P43119 and Q07869 transcripts peaked before day 12 and then declined and became nearly undetectable by day 17 , whereas Q03181 and P37231 transcript levels rose steadily from days 12 to 17 . Because the PPARs and the RXRs display different expression profiles , we suggest that different heterodimers may form and support distinct functions as development proceeds . Our results also underline the importance of Q16647 and Q03181 in the trophoblast and P43119 in the uterus , suggesting that DB01240 is of both uterine and trophoblastic origin and is involved in a complex signalling pathway at around the time of implantation in the ewe . [ Detection of a high-affinity prostaglandin I2 binding site in the human thyroid ] . This study is concerned with the identification and the pharmacological properties of P43119 binding sites on human thyroid membrane fractions . Scatchard analysis is not linear , revealing a high- and a low-affinity receptor binding site . ( 3 H ) DB01088 binding experiments were performed under various clinical conditions : in thyroid cancer the low-affinity binding sites disappear totally and the specific high-affinity binding sites are diminished according to the grade of differentiation of the cancer . An alteration in Bmax and Kd is also observed in cold nodules , in Hashimoto 's and Riedl 's thyroiditis and in hyperthyroidism , whereas hot nodules exhibit an increase in both the receptor subpopulations . The data provide evidence for specific DB01240 binding sites and support the suggestion of a direct regulatory key-role of DB01240 in thyroid intermediary metabolism . X-ray cross-complementing group 1 and thymidylate synthase polymorphisms might predict response to chemoradiotherapy in rectal cancer patients . PURPOSE : DB00544 -based chemoradiotherapy before total mesorectal excision is currently the standard treatment of Stage II and III rectal cancer patients . We used known predictive pharmacogenetic biomarkers to identify the responders to preoperative chemoradiotherapy in our series . METHODS AND MATERIALS : A total of 93 Stage II-III rectal cancer patients were genotyped using peripheral blood samples . The genes analyzed were X-ray cross-complementing group 1 ( P18887 ) , P07992 , P42898 , P00533 , Q12882 , and P04818 . The patients were treated with 225 mg/m(2)/d continuous infusion of 5-fluorouracil concomitantly with radiotherapy ( 50.4 Gy ) followed by total mesorectal excision . The outcomes were measured by tumor regression grade ( TRG ) as a major response ( TRG 1 and TRG 2 ) or as a poor response ( Q9Y512 , TRG4 , and TRG5 ) . RESULTS : The major histopathologic response rate was 47.3 % . P18887 G/G carriers had a greater probability of response than G/A carriers ( odds ratio , 4.18 ; 95 % confidence interval , 1.62-10.74 , p = .003 ) Patients with polymorphisms associated with high expression of thymidylate synthase ( 2R/3G , 3C/3G , and 3G/3G ) showed a greater pathologic response rate compared with carriers of low expression ( odds ratio , 2.65 ; 95 % confidence interval , 1.10-6.39 , p = .02 ) No significant differences were seen in the response according to P00533 , P07992 , MTHFR_C677 and MTHFR_A1298 expression . CONCLUSIONS : P18887 G/G and thymidylate synthase ( 2R/3G , 3C/3G , and 3G/3G ) are independent factors of a major response . Germline thymidylate synthase and P18887 polymorphisms might be useful as predictive markers of rectal tumor response to neoadjuvant chemoradiotherapy with 5-fluorouracil . Q99572 receptor-dependent intestinal afferent hypersensitivity in a mouse model of postinfectious irritable bowel syndrome . The DB00171 -gated P2X(7) receptor ( P2X(7)R ) was shown to be an important mediator of inflammation and inflammatory pain through its regulation of IL-1β processing and release . Trichinella spiralis-infected mice develop a postinflammatory visceral hypersensitivity that is reminiscent of the clinical features associated with postinfectious irritable bowel syndrome . In this study , we used P2X(7)R knockout mice ( P2X(7)R(-/-) ) to investigate the role of P2X(7)R activation in the in vivo production of IL-1β and the development of postinflammatory visceral hypersensitivity in the T. spiralis-infected mouse . During acute nematode infection , IL-1β-containing cells and P2X(7)R expression were increased in the jejunum of wild-type ( WT ) mice . Peritoneal and serum IL-1β levels were also increased , which was indicative of elevated IL-1β release . However , in the P2X(7)R(-/-) animals , we found that infection had no effect upon intracellular , plasma , or peritoneal IL-1β levels . Conversely , infection augmented peritoneal P01375 -α levels in both WT and P2X(7)R(-/-) animals . Infection was also associated with a P2X(7)R-dependent increase in extracellular peritoneal lactate dehydrogenase , and it triggered immunological changes in both strains . Jejunal afferent fiber mechanosensitivity was assessed in uninfected and postinfected WT and P2X(7)R(-/-) animals . Postinfected WT animals developed an augmented afferent fiber response to mechanical stimuli ; however , this did not develop in postinfected P2X(7)R(-/-) animals . Therefore , our results demonstrated that P2X(7)Rs play a pivotal role in intestinal inflammation and are a trigger for the development of visceral hypersensitivity . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass . P05181 epigenetic regulation in chronic , low-level toluene exposure : Relationship with oxidative stress and smoking habit . BACKGROUND : P05181 is a versatile phase I drug-metabolizing enzyme responsible for the biotransformation of most volatile organic compounds , including toluene . Human toluene exposure increases P05181 mRNA and modifies its activity in leucocytes ; however , epigenetic implications of this interaction have not been investigated . GOAL : To determine promoter methylation of P05181 and other genes known to be affected by toluene exposure . METHODS : We obtained venous blood from 24 tannery workers exposed to toluene ( mean levels : 10.86+/-7mg/m(3) ) and 24 administrative workers ( reference group , mean levels 0.21+/-0.02mg/m(3) ) all of them from the city of León , Guanajuato , México . After DNA extraction and bisulfite treatment , we performed PCR-pyrosequencing in order to measure methylation levels at promoter region of 13 genes . RESULTS : In exposed group we found significant correlations between toluene airborne levels and P05181 promoter methylation ( r=-.36 , p < 0.05 ) , as well as for P05231 promoter methylation levels ( r=.44 , p < 0.05 ) . Moreover , P05181 promoter methylation levels where higher in toluene-exposed smokers compared to nonsmokers ( p=0.009 ) . We also observed significant correlations for P05181 promoter methylation with P09211 and P00441 promoter methylation levels ( r=-.37 , p < 0.05 and r=-.34 , p < 0.05 respectively ) . CONCLUSION : These results highlight the importance of considering P05181 epigenetic modifications , as well as its interactions with other genes , as key factors for unraveling the sub cellular mechanisms of toxicity exerted by oxidative stress , which can initiate disease process in chronic , low-level toluene exposure . People co-exposed to toluene and tobacco smoke are in higher risk due to a possible P05181 repression . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation . Biophysical and pharmacological characterization of hypotonically activated chloride currents in cortical astrocytes . Rat cortical astrocytes regulate their cell volume in response to hypotonic challenge . This regulation is believed to depend largely on the release of chloride or organic osmolytes through anion channels . Using whole-cell recordings , we identified weakly outwardly rectifying chloride currents that could be activated in response to hypotonic challenge . These currents exhibited the following permeability sequence upon replacement of chloride in the bathing solution with various anions : I- > NO3- > Cl- > Gluc- > or = MeS- > Ise- . Interestingly , extracellular I- , albeit showing the greatest permeability , blocked the currents with an IC50 of approximately 50 mM . Currents were almost completely inhibited by 123 microM P16860 and partially inhibited by 200 microM niflumic acid or 200 microM DIDS . Additionally , the total number of Cl- ions effluxed through the hypotonically activated channels was markedly similar to the total solute efflux during volume regulation . We therefore propose the hypotonically activated chloride channel as a major contributor to volume regulation of astrocytes . To examine potential candidate chloride channel genes expressed by astrocytes , we employed RT-PCR to demonstrate the presence of transcripts for P51788 , 3 , 4 , 5 , and 7 , as well as for P21796 and P13569 in cultured astrocytes . Moreover , we performed immunostaining with antibodies against each of these channels and showed the strongest expression of P51788 and P51790 , strong expression of P51795 and P21796 , weak expression of P51798 and very weak expression of P51793 and P13569 . Intriguingly , although we found at least seven Cl- channel proteins from three different gene families in astrocytes , none appeared to be active in resting cells . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Splenic autonomic denervation increases inflammatory status but does not aggravate atherosclerotic lesion development . The brain plays a prominent role in the regulation of inflammation . Immune cells are under control of the so-called cholinergic anti-inflammatory reflex , mainly acting via autonomic innervation of the spleen . Activation of this reflex inhibits the secretion of proinflammatory cytokines and may reduce the development of atherosclerosis . Therefore , the aim of this study was to evaluate the effects of selective parasympathetic ( Px ) and sympathetic ( Sx ) denervation of the spleen on inflammatory status and atherosclerotic lesion development . Female P02649 *3-Leiden. P11597 mice , a well-established model for human-like lipid metabolism and atherosclerosis , were fed a cholesterol-containing Western-type diet for 4 wk after which they were subdivided into three groups receiving either splenic Px , splenic Sx , or sham surgery . The mice were subsequently challenged with the same diet for an additional 15 wk . Selective Px increased leukocyte counts ( i.e. , dendritic cells , B cells , and T cells ) in the spleen and increased gene expression of proinflammatory cytokines in the liver and peritoneal leukocytes compared with Sx and sham surgery . Both Px and Sx increased circulating proinflammatory cytokines IL-1β and P05231 . However , the increased proinflammatory status in denervated mice did not affect atherosclerotic lesion size or lesion composition . CONCLUSION : Predominantly selective Px of the spleen enhances the inflammatory status , which , however , does not aggravate diet-induced atherosclerotic lesion development . Human cytokines interleukin ( IL ) -3 and P05231 affect the growth and insulin binding of the unicellular organism Tetrahymena . Interleukin ( IL ) -3 and P05231 significantly increase the growth rate of the unicellular organism , Tetrahymena . The effect elicited by P08700 is long lasting as it was also detectable after 20 generations . Effect of P05231 was detectable as long as the substance was present in the cell culture . Pretreatment with P08700 did not enhance the proliferative response to subsequent P08700 treatment , but the second exposure to P08700 considerably depressed the active proliferation of Tetrahymena cells . However , a positive ' priming effect ' elicited by P05231 resulted in an increased growth rate following repeated P05231 stimulation . P01308 binding to the plasma membrane of Tetrahymena was increased by P05231 but not by P08700 after 24 hours , and this enhancement appeared even after one hour incubation . If the cells were pretreated with insulin , P05231 did not influence insulin binding , while an inhibition by P08700 was observed . These results direct attention to the similarities of actions induced by P08700 and P05231 at different levels of phylogeny probably due to the presence of cytokine receptor-like structures on this unicellular organism . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . Two types of prostacyclin receptor coupling to stimulation of adenylate cyclase and phosphatidylinositol hydrolysis in a cultured mast cell line , BNu-2cl3 cells . DB01240 ( DB01240 ) -mediated signal transduction was examined in interleukin 3 ( P08700 ) -dependent BNu-2cl3 mast cells . DB01088 , a stable DB01240 analogue , induced the accumulation of intracellular DB02527 and IP3 , and an increase in the intracellular Ca2+ concentration . Pretreatment of the cells with a protein kinase C activator , 12-O-tetradecanoyl phorbol 13-acetate , suppressed the iloprost-induced IP3 accumulation and Ca2+ mobilization , but inversely potentiated the DB02527 accumulation , suggesting that neither of these signal transduction pathways of iloprosts is the result of a secondary effect of activation of the other . Removal of P08700 from the culture medium reduced the iloprost-induced IP3 accumulation and Ca2+ mobilization , while it had no effect on the iloprost-induced DB02527 accumulation at all . These results taken together suggest that BNu-2cl3 cells express two types of P43119 ; one couples to stimulation of adenylate cyclase , its expression being independent of P08700 , while the other couples to phosphatidylinositol hydrolysis , its expression being dependent on P08700 .

MH_train_55

MH_train_55

MH_train_55

interacts_with DB06825?

multiple_choice

Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . A novel aptamer functionalized CuInS2 quantum dots probe for daunorubicin sensing and near infrared imaging of prostate cancer cells . In this paper , a novel daunorubicin ( DNR ) -loaded P15941 aptamer-near infrared ( Q9Y3T9 ) CuInS2 quantum dot ( DNR- P15941 -QDs ) conjugates were developed , which can be used as a targeted cancer imaging and sensing system . After the Q9Y3T9 CuInS2 QDs conjugated with the P15941 aptamer-( P01215 )7 , DNR can intercalate into the double-stranded CG sequence of the P15941 -QDs . The incorporation of multiple CG sequences within the stem of the aptamers may further increase the loading efficiency of DNR on these conjugates . DNR- P15941 -QDs can be used to target prostate cancer cells . We evaluated the capacity of P15941 -CuInS2 QDs for delivering DNR to cancer cells in vitro , and its binding affinity to P15941 -positive and P15941 -negative cells . This novel aptamer functionalized QDs bio-nano-system can not only deliver DNR to the targeted prostate cancer cells , but also can sense DNR by the change of photoluminescence intensity of CuInS2 QDs , which concurrently images the cancer cells . The quenched fluorescence intensity of P15941 -QDs was proportional to the concentration of DNR in the concentration ranges of 33-88 nmol L(-1) . The detection limit ( LOD ) for DNR was 19 nmol L(-1) . We demonstrate the specificity and sensitivity of this DNR- P15941 -QDs probe as a cancer cell imaging , therapy and sensing system in vitro . 25-Hydroxycholesterol is not a ligand for the orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) . The orphan nuclear receptor steroidogenic factor-1 ( Q13285 ) is involved in the transcriptional regulation of all the steroid hydroxylase genes , and also regulates the transcription of the genes for Müllerian Inhibitory substance ( P03971 ) , alpha subunit of glycoprotein hormone , LHbeta , oxytocin , P30968 , Q01718 , prolactin receptor , DAX-1 , and steroidogenic acute regulatory protein . Other members of the nuclear receptor gene family , including steroid hormone , thyroid hormone , retinoic acid , Q07869 , and vitamin D receptors must bind ligand to activate transcription , but Q13285 has been considered to be an orphan nuclear receptor because , when identified , it had no known ligand . A recent publication suggested that transcriptional regulation by Q13285 , expressed in a non-steroidogenic CV-1 cells , could be activated by oxysterols suggesting that these compounds could serve as natural ligands for Q13285 . We now demonstrate that 25-hydroxycholesterol , either added exogenously or synthesized endogenously in steroidogenic mouse Leydig MA-10 cells , did not act as a ligand for Q13285 , as it did not increase transcription from six different Q13285 -dependent DNA sequences . Furthermore , the abundance of these oxysterols in MA-10 cells was much less than concentrations needed for activation of Q13285 in CV-1 cells , indicating that Q13285 is not constitutively bound by ligand in MA-10 cells . Thus , in steroidogenic cells , transcriptional regulation of the steroid hydroxylase genes by Q13285 does not depend upon the presence of 25-hydroxycholesterol , and is not modified by its presence . DB00644 induction of extracellular-signal regulated kinase is blocked by inhibition of calmodulin . Our previous studies demonstrate that DB00644 -induced P29323 activation required influx of extracellular Ca2+ in alphaT3-1 and rat pituitary cells . In the present studies , we examined the hypothesis that calmodulin ( Cam ) plays a fundamental role in mediating the effects of Ca2+ on P29323 activation . Cam inhibition using W7 was sufficient to block DB00644 -induced reporter gene activity for the c-Fos , murine glycoprotein hormone alpha-subunit , and MAPK phosphatase ( MKP ) -2 promoters , all shown to require P29323 activation . Inhibition of Cam ( using a dominant negative ) was sufficient to block DB00644 -induced P29323 but not c-Jun N-terminal kinase activity activation . The Cam-dependent protein kinase ( CamK ) II inhibitor KN62 did not recapitulate these findings . DB00644 -induced phosphorylation of Q02750 and c-Raf kinase was blocked by Cam inhibition , whereas activity of phospholipase C was unaffected , suggesting that Ca2+/Cam modulation of the P29323 cascade potentially at the level of c-Raf kinase . Enrichment of Cam-interacting proteins using a Cam agarose column revealed that c-Raf kinase forms a complex with Cam . Reconstitution studies reveal that recombinant c-Raf kinase can associate directly with Cam in a Ca2+-dependent manner and this interaction is reduced in vitro by addition of W7 . Cam was localized in lipid rafts consistent with the formation of a Ca2+-sensitive signaling platform including the P30968 and c-Raf kinase . These data support the conclusion that Cam may have a critical role as a Ca2+ sensor in specifically linking Ca2+ flux with P29323 activation within the DB00644 signaling pathway . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Characterization of the human gonadotropin-releasing hormone receptor heterologously produced using the baculovirus/insect cell and the Semliki Forest virus systems . 1. Two eukaryotic viral systems , the baculovirus/insect cell and the Semliki Forest virus systems , were tested for heterologous expression of human gonadotropin-releasing hormone receptor ( GnRHR ) cDNA . 2 . An unmodified as well as a c-myc epitope-tagged human P30968 was produced in two insect cell lines ( Spodoptera frugiperda , Trichoplusia ni ) after infection with the respective recombinant baculoviruses . In both insect cell lines , the receptor was identified by immunoblot analysis as a triplet of bands between 35 and 40 kDa . After deglycosylation of the receptor the molecular mass decreased to 35 kDa . The P30968 was localized in membrane compartments within the infected insect cells . However , only in membranes of infected Trichoplusia ni insect cells could approximately 2000 receptors per cell be detected . 3 . Production of the P30968 in BHK cells using the Semliki Forest virus system resulted in approximately 50,000 receptors per cell . A maximal yield of 0.42 pmol/mg membrane protein was obtained 24 hr after electroporation of BHK cells with in vitro synthesized RNA . Binding of the antagonist [125I] DB00050 was saturable with a KD of 1.3 nM . The receptor produced in the BHK cells was further characterized by ligand displacement studies . The rank order of agonist and antagonist affinities was DB00050 > DB06825 > Antide > DB00644 . Human embryonic stem cells carrying an unbalanced translocation demonstrate impaired differentiation into trophoblasts : an in vitro model of human implantation failure . Carriers of the balanced translocation t(11;22) , the most common reciprocal translocation in humans , are at high risk of creating gametes with unbalanced translocation , leading to repeated miscarriages . Current research models for studying translocated embryos and the biological basis for their implantation failure are limited . The aim of this study was to elucidate whether human embryonic stem cells ( hESCs ) carrying the unbalanced chromosomal translocation t(11;22) can provide an explanation for repeated miscarriages of unbalanced translocated embryos . Fluorescent in situ hybridization and karyotype analysis were performed to analyze the t(11;22) in embryos during P52209 and in the derived hESC line . The hESC line was characterized by RT-PCR and FACS analysis for pluripotent markers . Directed differentiation to trophoblasts was carried out by bone morphogenetic protein 4 ( P12644 ) . Trophoblast development was analyzed by measuring β-hCG secretion , by β-hCG immunostaining and by gene expression of trophoblastic markers . We derived the first hESC line carrying unbalanced t(11;22) , which showed the typical morphological and molecular characteristics of a hESC line . Control hESCs differentiated into trophoblasts secreted increasing levels of β-hCG and concomitantly expressed the trophoblast genes , Q99626 , Q9H3D4 , P08729 , ERVW1 , P01215 , Q9NP62 , O43474 and P37231 . In contrast , differentiated translocated hESCs displayed reduced and delayed secretion of β-hCG concomitant with impaired expression of the trophoblastic genes . The reduced activation of trophoblastic genes may be responsible for the impaired trophoblastic differentiation in t(11;22)-hESCs , associated with implantation failure in unbalanced t(11;22) embryos . Our t(11;22) hESCs are presented as a valuable human model for studying the mechanisms underlying implantation failure . Association of germline p53 mutation with Q03164 segmental jumping translocation in treatment-related leukemia . Segmental jumping translocations are chromosomal abnormalities in treatment-related leukemias characterized by multiple copies of the P00519 and/or Q03164 oncogenes dispersed throughout the genome and extrachromosomally . Because gene amplification potential accompanies loss of wild-type p53 , we examined the p53 gene in a case of treatment-related acute myeloid leukemia ( t-AML ) with Q03164 segmental jumping translocation . The child was diagnosed with ganglioneuroma and embryonal rhabdomyosarcoma ( ERMS ) at 2 years of age . Therapy for ERMS included alkylating agents , P11387 and DNA topoisomerase II inhibitors , and local radiation . t-AML was diagnosed at 4 years of age . The complex karyotype of the t-AML showed structural and numerical abnormalities . Fluorescence in situ hybridization analysis showed multiple copies of the Q03164 gene , consistent with segmental jumping translocation . A genomic region including CD3 , Q03164 , and a segment of band 11q24 was unrearranged and amplified by Southern blot analysis . There was no family history of a cancer predisposing syndrome , but single-strand conformation polymorphism ( SSCP ) analysis detected identical band shifts in the leukemia , ganglioneuroma , ERMS , and normal tissues , consistent with a germline p53 mutation , and there was loss of heterozygosity in the ERMS and the t-AML . Sequencing showed a P01215 --> TGA nonsense mutation at codon 306 in exon 8 . The results of this analysis indicate that loss of wild-type p53 may be associated with genomic instability after DNA-damaging chemotherapy and radiation , manifest as a complex karyotype and gene amplification in some cases of t-AML . 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 . Polymorphisms associated with egg number at 300 days of age in chickens . We looked for variations that could be associated with chicken egg number at 300 days of age ( EN300 ) in seven genes of the hypothalamic-pituitary-gonadal axis , including gonadotrophin-releasing hormone-I ( DB00644 ) , P30968 ( GnRHR ) , neuropeptide Y ( P01303 ) , dopamine D2 receptor ( P14416 ) , vasoactive intestinal polypeptide ( P01282 ) , P01282 receptor-1 ( VIPR-1 ) , prolactin ( PRL ) , and the QTL region between 87 and 105 cM of the Z chromosome . Ten mutations in the seven genes were chosen to do marker-trait association analyses in a population comprising 1310 chickens , which were obtained from a company located in Guangdong Province of China . The C1704887T of VIPR-1 was found to have a highly significant association with EN300 . The T5841629C of P14416 and the C1715301T of VIPR-1 were significantly associated with EN300 . A highly significant association was also found between the C1704887T-C1715301T haplotypes of VIPR-1 and EN300 . H1H3 had the highest EN300 . Four PCR-RFLP variations in the candidate QTL region were selected to investigate their genetic effects on EN300 . The haplotypes of T32742468C-G32742603A in this region showed a highly significant association with EN300 . Bioinformatics analyses showed that both T32742468C and G32742603A were located in intron 1 of the SH3-domain P62993 -like 2 ( Q99962 ) gene . We conclude that five SNPs , including C1704887T and C1715301T of VIPR-1 , T5841629C of P14416 , and T32742468C and G32742603A of Q99962 , would be useful as markers for breeding to increase chicken EN300 . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Improvement of chloride transport defect by gonadotropin-releasing hormone ( DB00644 ) in cystic fibrosis epithelial cells . Cystic fibrosis ( CF ) , the most common autosomal recessive disease in Caucasians , is due to mutations in the P13569 gene . F508del , the most frequent mutation in patients , impairs P13569 protein folding and biosynthesis . The F508del- P13569 protein is retained in the endoplasmic reticulum ( ER ) and its traffic to the plasma membrane is altered . Nevertheless , if it reaches the cell surface , it exhibits a Cl(-) channel function despite a short half-life . Pharmacological treatments may target the F508del- P13569 defect directly by binding to the mutant protein or indirectly by altering cellular proteostasis , and promote its plasma membrane targeting and stability . We previously showed that annexine A5 ( AnxA5 ) directly binds to F508del- P13569 and , when overexpressed , promotes its membrane stability , leading to the restoration of some Cl(-) channel function in cells . Because Gonadotropin-Releasing Hormone ( DB00644 ) increases AnxA5 expression in some cells , we tested it in CF cells . We showed that human epithelial cells express DB00644 -receptors ( P30968 ) and that DB00644 induces an AnxA5 overexpression and an increased Cl(-) channel function in F508del- P13569 cells , due to an increased stability of the protein in the membranes . Beside the numerous physiological implications of the P30968 expression in epithelial cells , we propose that a topical use of DB00644 is a potential treatment in CF . Transcript and protein profiling identifies signaling , growth arrest , apoptosis , and NF-κB survival signatures following P01148 receptor activation . P01148 significantly inhibits proliferation of a proportion of cancer cell lines by activating P01148 receptor ( P30968 ) -G protein signaling . Therefore , manipulation of P30968 signaling may have an under-utilized role in treating certain breast and ovarian cancers . However , the precise signaling pathways necessary for the effect and the features of cellular responses remain poorly defined . We used transcriptomic and proteomic profiling approaches to characterize the effects of P30968 activation in sensitive cells ( HEK293- P30968 , SCL60 ) in vitro and in vivo , compared to unresponsive HEK293 . Analyses of gene expression demonstrated a dynamic response to the P01148 superagonist DB06825 . Early and mid-phase changes ( 0.5-1.0 h ) comprised mainly transcription factors . Later changes ( 8-24 h ) included a P01148 target gene , P01215 , and up- or downregulation of transcripts encoding signaling and cell division machinery . Pathway analysis identified altered MAPK and cell cycle pathways , consistent with occurrence of G(2)/M arrest and apoptosis . Nuclear factor kappa B ( NF-κB ) pathway gene transcripts were differentially expressed between control and DB06825 -treated SCL60 cultures . Reverse-phase protein and phospho-proteomic array analyses profiled responses in cultured cells and SCL60 xenografts in vivo during DB06825 anti-proliferation . Increased phosphorylated NF-κB ( p65 ) occurred in SCL60 in vitro , and p-NF-κB and IκBε were higher in treated xenografts than controls after 4 days DB06825 . NF-κB inhibition enhanced the anti-proliferative effect of DB06825 in SCL60 cultures . This study reveals details of pathways interacting with intense P30968 signaling , identifies potential anti-proliferative target genes , and implicates the NF-κB survival pathway as a node for enhancing P01148 agonist-induced anti-proliferation . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . P30968 and peritoneal plasmin activity . Most surgical procedures performed by obstetrician-gynecologists are associated with pelvic adhesions that cause subsequent serious sequelae , including small bowel obstruction , infertility , chronic pelvic pain , and difficulty in postoperative treatment , including complexity during subsequent surgical procedures . This study was conducted to determine if gonadotropin-releasing hormone analogues ( GnRHa ) affect the expressing tissue-type plasminogen activator ( t-PA ) and its inhibitor-1 ( P05121 ) in peritoneal cells in culture . Human peritoneal Met5A cells were used to examine the effects of GnRHa leuprolide , buserelin and goserelin on the levels of t-PA and PA-1 . Antigen concentrations were measured in conditioned media and cell lysates by real-time PCR and ELISA . P30968 ( GnRHR ) mRNA was determined by RT-PCR . GnRHR mRNA was detected in Met5A cells . Exposure of Met5A cells to GnRHa induced a rapid decrease of P05121 level in cultured medium but not in cell lysate ( protein and mRNA ) . These effects of GnRHa on P05121 were not associated with any changes in t-PA level . These results suggest that GnRHa may be an effective stimulator of local peritoneal fibrinolytic activity , as it decreases P05121 secretion in peritoneal Met5A cells by a mechanism linked to GnRHR . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Activin A augments DB00644 -mediated transcriptional activation of the mouse P30968 gene . The response of pituitary gonadotropes to DB00644 correlates directly with the concentration of DB00644 receptors ( GnRHRs ) on the cell surface , which is mediated in part at the level of GnRHR gene expression . We have previously localized DB00644 responsiveness in the mouse GnRHR ( mGnRHR ) gene promoter to two elements : activating protein-1 and sequence underlying responsiveness to DB00644 -1 . This study was designed to investigate potential synergy between DB00644 and activin A in transcriptional activation of the mGnRHR gene . In functional transfection studies using alphaT3-1 cells , DB00644 agonist stimulation of the mGnRHR gene promoter ( -765/+62 ) resulted in a 10.9-fold increase in activity , which was further increased 2-fold ( to 21.3-fold ) following activin pretreatment . Activin pretreatment alone had no effect . Deletion of region -387/-308 or mutation of a putative SMAD-binding element at -331/-324 ( 5'-GTCTAG[T]C-3' ) abrogated the augmented response to DB00644 in the presence of activin but not the response to DB00644 alone . Overexpression of Q15796 and P84022 along with Q13485 increased transcriptional activity of the mGnRHR gene , which was further increased by DB00644 agonist stimulation . These data demonstrate that activin augments DB00644 -mediated transcriptional activation of the mGnRHR gene and suggest that this effect may be mediated through SMAD transcription factors . Direct oral anticoagulants in acute coronary syndrome . Patients with acute coronary syndromes ( ACS ) require a specific antithrombotic therapy in the immediate and the post ACS phase . The current antithrombotic therapy in the acute phase of an ACS combines antiplatelet and anticoagulant drugs in order to reduce ischemic cardiovascular events . In the post ACS phase , dual antiplatelet therapy ( DAPT ; aspirin and a Q9H244 receptor antagonist ) is the current mainstay of antithrombotic treatment and is recommended in the guidelines of the major North American and European clinical cardiology associations ( DB00551 , ACC , and ESC ) . Recently , the addition of rivaroxaban , a low dose oral direct factor Xa inhibitor ( 2.5 mg twice daily ) , to DAPT ( aspirin plus second-generation Q9H244 inhibitor ) showed a significant reduction of cardiovascular and overall mortality in the major phase III clinical trial ATLAS ACS 2 TIMI 51 . This led to the approval of low-dose rivaroxaban in addition to aspirin and clopidogrel by the European Medicines Agency ( P15941 ) in 2013 . Other direct oral anticoagulants ( apixaban , dabigatran etexilate ) have also been assessed in phase II ( dabigatran etexilate ) and phase III ( apixaban ) post ACS clinical trials . In the studied dosing regimens , these drugs failed to show a net clinical benefit in addition to dual antiplatelet therapy . The major clinical phase II and III post ACS studies of direct oral anticoagulants are summarized and discussed in this article along with the concept of long-term anticoagulation for the secondary prevention of ischemic events after ACS and implications for the future of antithrombotic therapy in the current era of third-generation Q9H244 receptor inhibitors ( Prasugrel and DB08816 ) . P11387 is a cofactor for c-Jun in the regulation of epidermal growth factor receptor expression and cancer cell proliferation . P11387 ( Topo I ) is a molecular target for the anticancer agent topotecan in the treatment of small cell lung cancer and ovarian carcinomas . However , the molecular mechanisms by which topotecan treatment inhibits cancer cell proliferation are unclear . We describe here the identification of Topo I as a novel endogenous interaction partner for transcription factor c-Jun . Reciprocal coimmunoprecipitation analysis showed that Topo I and c-Jun interact in transformed human cells in a manner that is dependent on JNK activity . c-Jun target gene epidermal growth factor receptor ( P00533 ) was identified as a novel gene whose expression was specifically inhibited by topotecan . Moreover , Topo I overexpression supported c-Jun-mediated reporter gene activation and both genetic and chemical inhibition of c-Jun converted cells resistant to topotecan-elicited P00533 downregulation . DB01030 -elicited suppression of proliferation was rescued by exogenously expressed P00533 . Furthermore , we demonstrate the cooperation of the JNK-c-Jun pathway , Topo I , and P00533 in the positive regulation of O75794 cell proliferation . Together , these results have identified transcriptional coactivator Topo I as a first endogenous cofactor for c-Jun in the regulation of cell proliferation . In addition , the results of the present study strongly suggest that inhibition of P00533 expression is a novel mechanism by which topotecan inhibits cell proliferation in cancer therapy . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . The coffee constituent chlorogenic acid induces cellular DNA damage and formation of topoisomerase I- and II-DNA complexes in cells . Chlorogenic acid ( P01215 ) is a plant polyphenol with known antioxidant properties . Although some studies suggest that P01215 has anticancer properties , others indicate that this dietary constituent may cause DNA damage and induce carcinogenic effects . Because P01215 is widely consumed in the form of coffee , it is important to further evaluate the putative DNA-damaging activity of P01215 . Here we have employed two standard techniques commonly used for DNA damage detection ( the comet assay and the γ- P16104 focus assay ) and observed that P01215 ( 0.5-5 mM ) induces DNA damage in normal and cancer cells . We report for the first time that P01215 induces high levels of topoisomerase I- and topoisomerase II-DNA complexes in cells ( TARDIS assay ) . P04040 pretreatment abolished the formation of these topoisomerase-DNA complexes and reduced the cytotoxic activity of P01215 , therefore indicating that hydrogen peroxide plays an important role in these activities . Lung cancer cells ( A549 ) were more sensitive than normal lung fibroblasts ( MRC5 ) to the cytotoxic activity of P01215 , supporting previous findings that P01215 may induce selective killing of cancer cells . Taking into consideration our results and the pharmacokinetic profile of P01215 , the possible cancer preventive , carcinogenic and therapeutic potential of this dietary agent are discussed . Population pharmacokinetic study of memantine : effects of clinical and genetic factors . BACKGROUND AND OBJECTIVE : Memantine , a frequently prescribed anti-dementia drug , is mainly eliminated unchanged by the kidneys , partly via tubular secretion . Considerable inter-individual variability in plasma concentrations has been reported . We aimed to investigate clinical and genetic factors influencing memantine disposition . METHODS : A population pharmacokinetic study was performed including data from 108 patients recruited in a naturalistic setting . Patients were genotyped for common polymorphisms in renal cation transporters ( O15245 /2/5 , Q96FL8 , P08183 ) and nuclear receptors ( O75469 , Q14994 , RXR , Q07869 ) involved in transporter expression . RESULTS : The average clearance was 5.2 L/h with a 27 % inter-individual variability ( percentage coefficient of variation ) . Glomerular filtration rate ( p = 0.007 ) and sex ( p = 0.001 ) markedly influenced memantine clearance . O75469 rs1523130 was identified as the unique significant genetic covariate for memantine clearance ( p = 0.006 ) , with carriers of the O75469 rs1523130 CT/TT genotypes presenting a 16 % slower memantine elimination than carriers of the CC genotype . CONCLUSION : The better understanding of inter-individual variability of memantine disposition might be beneficial in the context of individual dose optimization . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . Effects of gonadoliberin analogue triptorelin on the pituitary-testicular complex in neonatal rats . DB06825 , a synthetic analogue of neurohormone gonadoliberin ( gonadotropin-releasing hormone , DB00644 ) administered daily to rats on postnatal days 5-7 suppressed the expression of P30968 in the pituitary gland , but did not change functioning of the pituitary-testicular complex . Administration of triptorelin on postnatal days 12-14 ( i.e. during the formation of pulsatile pattern of DB00644 secretion and increasing levels of its mRNA receptor in the pituitary gland ) had no effect on receptor expression , but increased the levels of luteinizing hormone mRNA in the pituitary gland and the weight of testes . At that time , blood levels of testosterone were lowered , which indicated disturbed pulsatile pattern of DB00644 secretion . Luteinizing Hormone-Releasing Hormone ( P01148 ) -I antagonist cetrorelix inhibits myeloma cell growth in vitro and in vivo . The objective of this study was to determine the effects of an luteinizing hormone-releasing hormone ( P01148 ) -I antagonist , DB00050 , on human multiple myeloma ( MM ) cells and to elucidate the mechanisms of action . We showed that P01148 -I and P22888 -I genes were expressed in MM cell lines and primary MM cells . Treatment with DB00050 inhibited growth and colony-forming ability of myeloma cells , including cell lines resistant to arsenic trioxide , bortezomib , or lenalidomide . DB00050 induced apoptosis in myeloma cells including primary myeloma cells . In addition , DB00050 inhibited the growth of human myeloma cells xenografted into mice without any apparent side effects . DB00050 downregulated the nuclear factor-kappa B ( NF-κB ) pathway activity and the expression of cytokines , including interleukin 6 , insulin-like growth factor 1 , P15692 , and stromal-derived factor 1 , important for myeloma cell growth and survival in myeloma cells and/or marrow stromal cells from myeloma patients . DB00050 decreased the phosphorylation of extracellular signal regulated kinase 1/2 and P40763 in myeloma cells , two crucial pathways for myeloma cells growth and survival . Moreover , the expression of P38936 and p53 was increased , whereas that of antiapoptotic proteins Bcl-2 and Bcl-x(L) was reduced by DB00050 . Our findings indicate that DB00050 induces cytotoxicity in myeloma cells through various mechanisms and provide a rationale for investigating DB00050 for the treatment of MM . Selective cleavage of ErbB4 by G-protein-coupled gonadotropin-releasing hormone receptor in cultured hypothalamic neurons . DB00644 ( DB00644 ) is secreted from hypothalamic neurons ( DB00644 neurons ) . DB00644 neurons have a P30968 belonging to the G-protein-coupled receptors . The stimulation of this receptor activates extracellular signal-regulated kinase ( P29323 ) . In the present study , we found that epidermal growth factor receptor ( P00533 ) and ErbB4 were expressed in immortalized DB00644 neurons ( GT1-7 cells ) . AG1478 , a relatively specific inhibitor of the ErbB family , and small interfering RNA ( siRNA ) for ErbB4 inhibited the DB00644 -induced activation of P29323 in GT1-7 cells , suggesting that P00533 and ErbB4 were necessary for the activation . In addition , DB00644 induced the cleavage of ErbB4 and accumulation of an 80-kDa fragment . After treatment of the cells with 50 nM DB00644 for 5 min , about 80 % of ErbB4 was cleaved . Biotinylation of cell surface proteins revealed that more than 70 % of the cell surface ErbB4 was cleaved by DB00644 treatment . A higher concentration and longer treatment were necessary for DB00644 to induce ErbB4 cleavage than P29323 activation . TAPI-2 , an inhibitor of tumor necrosis factor-α-converting enzyme ( P78536 ) , and siRNA for P78536 inhibited the cleavage of ErbB4 , suggesting that P78536 was involved . After ErbB4 cleavage , the activation of P29323 by neuregulin 1 was almost completely inhibited . These results suggest that the down-regulation of ErbB4 expression is induced by G-protein-coupled receptor stimulation . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . DB01281 inhibits effector T cells through regulatory T cells and TGF-β . The P10747 costimulatory receptor is a critical regulator of T cell function , making it an attractive therapeutic target for the treatment of immune-mediated diseases . DB01281 , now approved for use in humans , prevents naive T cell activation by binding to P33681 proteins and blocking engagement of P10747 . However , DB01281 suppresses inflammation even if administered when disease is established , suggesting alternative mechanisms . We identified a novel , P10747 -independent mechanism by which DB01281 inhibits activated T cells . We show that in vitro , DB01281 synergizes with NO from bone marrow-derived macrophages to inhibit T cell proliferation . Depletion of regulatory T cells ( Tregs ) or interference with TGF-β signaling abrogated the inhibitory effect of DB01281 . Parallel in vivo experiments using an allergic airway inflammation model demonstrated that this novel mechanism required both macrophages and regulatory T cells . Furthermore , DB01281 was ineffective in P84022 -deficient mice , supporting a requirement for TGF-β signaling . Thus , in addition to preventing naive T cells from being fully activated , DB01281 can turn off already activated effector T cells by an NO/regulatory T cell/TGF-β-dependent pathway . This mechanism is similar to cell-extrinsic effects of endogenous P16410 and may be particularly important in the ability of DB01281 to treat chronic inflammatory disease . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Impact of aromatic residues within transmembrane helix 6 of the human gonadotropin-releasing hormone receptor upon agonist and antagonist binding . To investigate the impact of aromatic residues within transmembrane helix 6 ( TMH6 ) of the human gonadotropin-releasing hormone receptor ( P30968 ) on agonist and antagonist binding , residues Y(283) , Y(284) , W(289) , Y(290) , W(291) , and F(292) were exchanged to alanine and analyzed comprehensively in functional reporter gene and ligand binding assays . Whereas receptor mutants Y(283)A , Y(284)A , and W(291)A were capable of neither ligand binding nor signal transduction , mutants W(289)A , Y(290)A , and F(292)A were functional : the F(292)A mutant behaved like wild-type receptor , while mutants W(289)A and Y(290)A differentiated between agonistic and antagonistic ligands . On the basis of the high-resolution X-ray structure of bovine rhodopsin as well as available data on P30968 mutants , models for ligand-receptor interactions are proposed . The model for D- DB00150 (6)- DB00644 ( DB06825 ) binding , representing a superagonistic ligand , is in full accordance to available data . Furthermore , new interactions are proposed : pGlu(1) interacts with N(212) in transmembrane helix 5 , DB00135 (5) with Y(290) , and D- DB00150 (6) with W(289) . The binding behavior of mutants W(289)A and Y(290)A corresponds to the proposed binding model for the antagonist DB00050 . In summary , our data as presented indicate that Y(290) plays a key function in agonist but not antagonist binding .

MH_train_56

MH_train_56

MH_train_56

interacts_with DB00921?

multiple_choice

[ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Preliminary evidence of ethnic divergence in associations of putative genetic variants for methamphetamine dependence . Research into the biological processes that increase susceptibility to methamphetamine dependence has been conducted primarily in Asian populations . Using a case-control design this study 's purpose was to explore , among a population of methamphetamine-dependent Caucasians , six putative single nucleotide polymorphisms previously found to be associated with methamphetamine dependence in Asian populations . A total of 193 non-psychotic males ( 117 methamphetamine-dependent and 76 controls ) were genotyped for variants located in six genes ( P31749 , P32121 , P23560 , P21964 , P09211 , P35372 ) . Genotypic and allelic frequencies , odds ratios , and 95 % confidence intervals were calculated . None of the putative gene associations was significantly replicated in our sample of Caucasian men . Effect size comparisons suggest a trend toward allelic divergence for arrestin beta 2 ( P32121 ) and glutathione S-transferase P1 ( P09211 ) and allelic convergence for brain-derived neurotrophic factor ( P23560 ) . Results provide preliminary support for further exploration and validation of candidate single nucleotide polymorphisms ( SNPs ) for methamphetamine ( METH ) dependence reported among Asian populations across other ethnic/ancestral groups . Novel camptothecin analogues that circumvent Q9UNQ0 -associated drug resistance in human tumor cells . DB00762 ( 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin ; CPT-11 ) is a widely used potent antitumor drug that inhibits mammalian P11387 ( Topo I ) ; however , overexpression of Q9UNQ0 ( Q9UNQ0 / Q9UNQ0 / Q9UNQ0 ) can confer cancer cell resistance to SN-38 , the active form of CPT-11 . We have recently demonstrated that plasma membrane vesicles prepared from Q9UNQ0 -overexpressing PC-6/ Q8WUX1 -5H cells transported SN-38 and its glucuronide conjugate in an DB00171 -dependent manner ( Nakatomi et al. , Biochem Biophys Res Commun 2001;288:827-32 ) . In the present study , we have characterized a total of 14 new camptothecin ( CPT ) analogues with respect to both the inhibition of Topo I and the substrate specificity of Q9UNQ0 . All of the tested CPT analogues , which have different substitutions at positions 10 and 11 , strongly inhibited the Topo I activity in a cell-free system , as did SN-38 . Their antitumor activities in the SN-38-resistant PC-6/ Q8WUX1 -5H2 cell line greatly varied , however , being correlated with intracellular accumulation levels . We have examined DB00171 -dependent transport of those CPT analogues by using plasma membrane vesicles prepared from both PC-6/ Q8WUX1 -5H2 cells and Q9UNQ0 -transfected P29320 -293 cells . Based on the substrate specificity of Q9UNQ0 thus evaluated , it is strongly suggested that CPT analogues with high polarity are good substrates for Q9UNQ0 and are therefore effectively extruded from cancer cells . In this context , to circumvent Q9UNQ0 -associated drug resistance , low-polarity CPT analogues are considered to be potent lead compounds . The present study provides a practical approach to discover new CPT-based drugs for the chemotherapy of drug-resistant human cancer . DB01393 restores the inhibition of DB00094 -induced follicular development and steroidogenesis by tumor necrosis factor-alpha through peroxisome proliferator-activated receptor-gamma pathway in an in vitro mouse preantral follicle culture . We recently reported that bezafibrate , a lipid-lowering drug of the fibrate class , administered in addition to clomiphene citrate ( CC ) successfully induced ovulation in CC-resistant polycystic ovary syndrome ( PCOS ) patients . We hypothesized that bezafibrate may directly affect ovarian follicle development . P01308 resistance and compensatory hyperinsulinemia are important for the pathogenesis of PCOS . In this study , we first examined the effects of tumor necrosis factor-alpha ( P01375 ) , which plays a role in insulin resistance , on follicle development by using the follicle culture system . P01375 significantly inhibited follicle-stimulating hormone ( DB00094 ) -induced follicle development , 17beta-estradiol ( E2 ) secretion , and ovulation rate in a dose-dependent manner . We then examined whether bezafibrate treatment could rescue the inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 . DB01393 treatment rescued inhibition of follicle development , secretion of E2 , and ovulation rate by P01375 . We examined the expression of peroxisome proliferator-activated receptor ( Q07869 ) subtypes in mouse preantral follicles . As the protein expression of only P37231 was observed in mouse preantral follicles , we examined whether bezafibrate could affect follicle development and steroidogenesis through P37231 pathways . Treatment with GW1929 , a selective P37231 agonist , restored inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 , whereas treatment with GW9662 , a selective P37231 antagonist , canceled the restorative effects of bezafibrate . Collectively , the results in this study suggest that bezafibrate may directly exhibit a restorative effect on the inhibition of ovarian follicle development and steroidogenesis by P01375 through the P37231 pathway . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . The P35372 promotes opioid and growth factor-induced proliferation , migration and Epithelial Mesenchymal Transition ( EMT ) in human lung cancer . Recent epidemiologic studies implying differences in cancer recurrence based on anesthetic regimens raise the possibility that the mu opioid receptor ( MOR ) can influence cancer progression . Based on our previous observations that overexpression of MOR in human non-small cell lung cancer ( NSCLC ) cells increased tumor growth and metastasis , this study examined whether MOR regulates growth factor receptor signaling and epithelial mesenchymal transition ( EMT ) in human NSCLC cells . We utilized specific siRNA , shRNA , chemical inhibitors and overexpression vectors in human H358 NSCLC cells that were either untreated or treated with various concentrations of DAMGO , morphine , fentanyl , P01133 or IGF . Cell function assays , immunoblot and immunoprecipitation assays were then performed . Our results indicate MOR regulates opioid and growth factor-induced P01133 receptor signaling ( Src , Gab-1 , PI3K , Akt and P40763 activation ) which is crucial for consequent human NSCLC cell proliferation and migration . In addition , human NSCLC cells treated with opioids , growth factors or MOR overexpression exhibited an increase in snail , slug and vimentin and decrease ZO-1 and claudin-1 protein levels , results consistent with an EMT phenotype . Further , these effects were reversed with silencing ( shRNA ) or chemical inhibition of MOR , Src , Gab-1 , PI3K , Akt and P40763 ( p < 0.05 ) . Our data suggest a possible direct effect of MOR on opioid and growth factor-signaling and consequent proliferation , migration and EMT transition during lung cancer progression . Such an effect provides a plausible explanation for the epidemiologic findings . Autoantibodies against four kinds of neurotransmitter receptors in psychiatric disorders . There is a hypothesis that autoimmune abnormalities in neurotransmitter receptors might cause some psychiatric disorders . Using a sensitive radioligand assay , we detected serum autoantibodies to recombinant human muscarinic cholinergic receptor 1 ( P11229 , 34.4 % ) , mu-opioid receptor ( P35372 , 13.1 % ) , P08908 ( P08908 , 7.4 % ) , and dopamine receptor D2 ( P14416 , 4.9 % ) in 122 psychiatric patients . Positive antibodies to P11229 were found in 34.1 % , 34.9 % , 33.3 % , and 9.1 % of patients with schizophrenic disorders ( n=44 ) , mood disorders ( n=63 ) , other psychiatric disorders ( n=15 ) and autoimmune diseases ( n=33 ) , respectively . All three patients with neuroleptic maliganant syndrome had high activities of autoantibodies to P11229 , P35372 , and/or P08908 . Our data suggest that autoimmunity to neurotransmitter receptors might be associated with the induction of psychiatric symptoms and have some relation to neuroleptic malignant syndrome . Microglial activation , increased P01375 and P31645 expression in the prefrontal cortex define stress-altered behaviour in mice susceptible to anhedonia . A chronic stress paradigm comprising exposure to predation , tail suspension and restraint induces a depressive syndrome in C57BL/6J mice that occurs in some , but not all , animals . Here , we sought to extend our behavioural studies to investigate how susceptibility ( sucrose preference < 65 % ) or resilience ( sucrose preference > 65 % ) to stress-induced anhedonia affects the 5HT system and the expression of inflammation-related genes . All chronically stressed animals , displayed increased level of anxiety , but susceptible mice exhibited an increased propensity to float in the forced swim test and demonstrate hyperactivity under stressful lighting conditions . These changes were not present in resilient or acutely stressed animals . Compared to resilient animals , susceptible mice showed elevated expression of tumour necrosis factor alpha ( P01375 ) and the 5-HT transporter ( P31645 ) in the pre-frontal area . Enhanced expression of 5HT(2A) and P23219 in the pre-frontal area was observed in all stressed animals . In turn , indoleamine-2,3-dioxygenase ( P14902 ) was significantly unregulated in the raphe of susceptible animals . At the cellular level , increased numbers of Iba-1-positive microglial cells were also present in the prefrontal area of susceptible animals compared to resilient animals . Consequently , the susceptible animals display a unique molecular profile when compared to resilient , but anxious , animals . Unexpectedly , this altered profile provides a rationale for exploring anti-inflammatory , and possibly , P01375 -targeted therapy for major depression . Linkage assignment of eleven genes to the porcine genome . We report comparative linkage mapping of eleven genes in the swine genome by RFLP analysis . These genes include : Acid phosphatase type 5 ( P13686 ) , Cholecystokinin Type B Receptor ( P32239 ) , Antibiotic Peptide ( P49913 ) , P01308 -like Growth Factor 1 Receptor ( P08069 ) , Integrin Alpha M ( P11215 ) , Integrin Beta 2 ( ITGbeta2 ) , Opioid Receptor Mu-1 ( P35372 ) , Pro-hormone Converter ( PC1/3 ) , DB00162 Binding Q12988 ( P10745 ) , Ribosomal DNA ( RNR1 ) , and Zona Pellucida Glycoprotein 1 ( P60852 ) . The P32239 and ITGbeta2 loci define the ends of the linkage groups on Chromosomes ( Chro ) ( SSC ) 9p and 13qter , respectively . P35372 phosphorylation , desensitization , and ligand efficacy . Mu opioid receptors are subject to phosphorylation and desensitization through actions of at least two distinct biochemical pathways : agonist-dependent mu receptor phosphorylation and desensitization induced by a biochemically distinct second pathway dependent on protein kinase C activation ( 1 ) . To better understand the nature of the agonist-induced mu receptor phosphorylation events , we have investigated the effects of a variety of opiate ligands of varying potencies and intrinsic activities on mu receptor phosphorylation and desensitization . Exposure to the potent full agonists sufentanil , dihydroetorphine , etorphine , etonitazine , and [ D-Ala2 , MePhe4 , Glyol5 ] enkephalin ( DAMGO ) led to strong receptor phosphorylation , while methadone , l-alpha-acetylmethadone ( DB01227 ) , morphine , meperidine , DADL , beta-endorphin(1-31) , enkephalins , and dynorphin A(1-17) produced intermediate effects . The partial agonist buprenorphine minimally enhanced receptor phosphorylation while antagonists failed to alter phosphorylation . DB00921 and full antagonists each antagonized the enhanced mu receptor phosphorylation induced by morphine or DAMGO . The rank order of opiate ligand efficacies in producing mu receptor-mediated functional desensitization generally paralleled their rank order of efficacies in producing receptor phosphorylation . Interestingly , the desensitization and phosphorylation mediated by methadone and DB01227 were disproportionate to their efficacies in two distinct test systems . This generally good fit between the efficacies of opiates in mu receptor activation , phosphorylation , and desensitization supports the idea that activated receptor/agonist/G-protein complexes and/or receptor conformational changes induced by agonists are required for agonist-induced mu receptor phosphorylation . Data for methadone and DB01227 suggest possible contribution from their enhanced desensitizing abilities to their therapeutic efficacies . DB00619 inhibition effect on KITAsn822Lys-mediated signal transduction cascade . OBJECTIVE : Alterations in growth factor signaling pathways may be a frequent collaborating event in Q01196 - Q06455 -mediated leukemogenesis . Gain-of-function P10721 receptor mutations have been reported in adult AML patients , especially those with core binding factor leukemia ( CBFL ) . We have previously reported a new gain-of-function P10721 (Asn822Lys) mutation that is constitutively expressed in the Kasumi-1 CBFL cell line , and has recently been described in two childhood AML patients . To explore the molecular basis of the effects of this mutation in the appropriate context of hemopoietic dysregulation , we investigated P10721 downstream signaling in the Kasumi-1 cell line by means of DB00619 ( Imatinib , Gleevec ) pharmacological inhibition . MATERIALS AND METHODS : We investigated P10721 (Asn822Lys) mutant-initiated signaling in Kasumi-1 cell line , and characterized the inhibitory effect of the DB00619 protein tyrosine kinase inhibitor on downstream signaling . RESULTS : The use of DB00619 -mediated inhibition impaired the tyrosine phosphorylation of P10721 (Asn822Lys) and its association with the p85 subunit of phosphatidylinositol 3'-kinase ( p85PI3K ) . The downstream constitutive phosphorylation of P45983 /2 and P40763 was also significantly inhibited , but DB00619 had no effect on the constitutive activation of Akt , thus suggesting that it is due to other signaling in Kasumi-1 cells . DB00619 inhibited the P10721 -mediated proliferation of Kasumi-1 cells in a dose-dependent manner . CONCLUSIONS : These findings show the role of PI3K in P10721 (Asn822Lys)-mediated constitutive activation through the Akt-independent downstream signaling pathway of JNK , and also demonstrate the mutant 's susceptibility to DB00619 , which may therefore have therapeutic potential in CBFL patients with susceptible P10721 mutations . Inhibition of Akt/ P31749 by a P35354 inhibitor induces apoptosis in gastric cancer cells . BACKGROUND/AIM : Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs . This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway . METHODS : Two gastric cancer cell lines , AGS and MKN28 , were treated with SC236 and assessed for cell growth and apoptosis . The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B ( Akt/ P31749 ) pathways and their downstream signalings were studied in the AGS cell line . RESULTS : SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase , but down-regulated Akt/ P31749 . The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed , while the constitutively active form of Akt/ P31749 was able to block SC236-induced apoptosis . SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases . CONCLUSION : One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c . Predictive model for risk of severe gastrointestinal toxicity following chemotherapy using patient immune genetics and type of cancer : a pilot study . PURPOSE : Severe chemotherapy-induced gastrointestinal toxicity ( CIGT ) is common and results in treatment delays , dose reductions , and potential premature treatment discontinuation . Currently , there is no diagnostic marker to predict CIGT . Proinflammatory cytokines , produced via toll-like receptor signaling , are key mediators of this toxicity . Hence , this pilot study investigated the association between immune genetic variability and severe CIGT risk . METHODS : Genomic DNA from 34 patients ( 10 with severe CIGT ) who had received 5-fluoruracil-based chemotherapy regimens was analyzed for variants of IL-1B , P60568 , P05231 , IL-6R , P22301 , P01375 , TGF-b , O60603 , O00206 , Q9Y6Y9 , Q99836 , P23560 , CRP , ICE , and P35372 . Multivariate logistic regression created a prediction model of severe CIGT risk . RESULTS : There were no significant differences between patients with and without severe CIGT with regards to age , sex , type of cancer , or chemotherapy treatment regimens . The prediction model of severe CIGT risk included O60603 and P01375 genetic variability and cancer type ( colorectal and gastric ) . This prediction model was both specific and sensitive , with a receiver operator characteristic area under the curve of 87.3 % . CONCLUSIONS : This is the first report of immune genetic variability , together with cancer type , being predictive of severe CIGT risk . These outcomes are being validated in a larger patient population . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . The P04035 inhibitor pravastatin stimulates insulin secretion through organic anion transporter polypeptides . The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor pravastatin has been reported to have a beneficial effect on reducing the new onset of diabetes as well as lowering plasma lipids . Because pravastatin is a water-soluble organic anion , it can not easily penetrate the lipid bilayer of the cell membrane . As the precise mechanisms of the effect of pravastatin on glucose metabolism and diabetes have not been clarified , we examined the roles of the organic anion transporter family on pravastatin-treated islet and adipocyte functions . Rat oatp1/slco1a1 , oatp2/slco1a4 and oatp3/slco1a5 were expressed in the pancreas , and rat oatp3/slco1a5 was also detected in rat insulinoma cell line P01308 -1e . DB00175 was transported not only by oatp1/slco1a1 and oatp2/slco1a4 , but also by rat oatp3/slco1a5 . DB00175 uptake into P01308 -1e cells was detected and this transport was inhibited by sulfobromophthalein and rifampicin , both of which are known to inhibit oatp family-mediated uptake . In addition , pravastatin enhanced the glucose-stimulated insulin secretion from P01308 -1e cells . When fat-loaded db/db mice were treated with pravastatin , glucose intolerance and insulin resistance were prevented . In addition , insulin secretion from isolated islets was enhanced by pravastatin . These data suggest that pravastatin has pleiotropic effects on islets through membrane transport under high fat/glucose conditions .

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The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . PMA-induced dissociation of P13010 from the promoter causes transcriptional up-regulation of histamine H(1) receptor . DB11320 H(1) receptor ( P35367 ) gene is up-regulated in patients with allergic rhinitis , and its expression level strongly correlates with the severity of symptoms . However , the mechanism underlying this remains unknown . Here we report the mechanism of P35367 gene up-regulation . The luciferase assay revealed the existence of two promoter regions , A and B1 . Two AP-1 and one Ets-1 bound to region A , while P13010 , P12956 , and P09874 bound to region B1 . P13010 was responsible for DNA binding and poly(ADP-ribosyl)ated in response to phorbol-12-myristate-13-acetate stimulation , inducing its dissociation from region B1 that is crucial for promoter activity . Knockdown of P13010 gene enhanced up-regulation of P35367 gene expression . Experiments using inhibitors for MEK and P09874 indicate that regions A and B1 are downstream regulatory elements of the PKCδ/ P29323 / P09874 signaling pathway . Data suggest a novel mechanism for the up-regulation of P35367 gene expression . Clinical and genetic factors associated with nausea and vomiting in cancer patients receiving opioids . BACKGROUND : This study investigates whether demographical , disease-related and genetic factors contribute to inter-individual differences in nausea and vomiting among patients receiving opioids for cancer pain . METHODS : Cancer patients receiving opioids were included from 17 centres in 11 European countries . Intensities of nausea and vomiting were reported by 1579 patients on four-point categorical scales . In stratified regression models including demographical and disease-related factors as covariates , 96 single nucleotide polymorphisms ( SNPs ) in 16 candidate genes related to opioid- or nausea/vomiting signalling pathways ( P08183 , P35372 , P41145 , P32121 , P42226 , P21964 , P20309 , P08912 , P35367 , P14416 , P35462 , P25103 , P46098 , O95264 , Q8WXA8 , P21554 ) were analysed for association with nausea and vomiting . FINDINGS : Age , body mass index , Karnofsky Performance Status , gender , use of antiemetics , type of opioid , type of cancer and eight SNPs were associated with the inter-individual differences in nausea and vomiting among cancer patients treated with opioids ( p < 0.01 ) . The SNPs were rs1176744 , rs3782025 and rs1672717 in O95264 ; rs165722 , rs4680 and rs4633 in P21964 ; rs10802789 and rs685550 in P20309 . Only the SNP rs1672717 in O95264 passed the Benjamini-Hochberg criterion for a 10 % false discovery rate . INTERPRETATION : Clinical characteristics and SNPs within the O95264 , P21964 and P20309 genes may be associated with the variability in nausea and vomiting among cancer patients receiving opioids . This knowledge may help to identify patients at particular risk for nausea and vomiting during treatment with opioids for cancer pain . FcεRI stimulation promotes the differentiation of histamine receptor 1-expressing inflammatory macrophages . BACKGROUND : Monocyte differentiation into dendritic cells or macrophages and recruitment to peripheral organs in chronic inflammatory diseases are directed by allergen challenge via FcεRI as well as the nature of soluble factors in the microenvironment . High-affinity receptor for IgE stimulation of effector cells results in the release of histamine , which acts on various histamine receptors ( HR ) 1-4 , expressed by immune cells . METHODS : We examined the effect of FcεRI stimulation of human monocytes on P35367 expression and function of differentiating cells . The mRNA levels of P35367 , P25021 and histidine decarboxylase of differentiating cells were detected by quantitative real-time PCR . Expression of CD1c , CD11c , P34810 and Q86VB7 was detected by flow cytometry . Amount of histamine , P05231 and IL-12p70 in the cell culture was measured with the help of cytometric bead arrays or ELISA assays . Numbers of P35367 -expressing macrophages were evaluated by immunofluorescence double staining of P34810 and P35367 on human skin sections . RESULTS : We demonstrated that FcεRI stimulation promotes the generation of P35367 -expressing macrophage-like cells with enhanced histamine biosynthesis and P35367 -mediated proinflammatory properties . Supporting our in vitro findings , high numbers of P35367 -expressing P34810 (pos) macrophages were detected in the dermis of atopic dermatitis ( AD ) skin lesions . CONCLUSION : Our observations point to a close histamine-/HR-mediated activation of dermal macrophages , leading to modified cell differentiation and responsiveness via P35367 , which might contribute to the aggravation of allergic skin inflammation in AD . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . Granulocyte macrophage-colony stimulating factor increases the expression of histamine and histamine receptors in monocytes/macrophages in relation to arteriosclerosis . OBJECTIVE : To study the effect of granulocyte macrophage-colony-stimulating factor ( GM- P04141 ) on histamine metabolism in arteriosclerosis , the expression of histidine decarboxylase ( HDC ; histamine-producing enzyme ) , histamine receptors 1 and 2 ( P35367 and P25021 ) , and GM- P04141 was investigated in human and mouse arteriosclerotic carotid arteries . Furthermore , the molecular mechanisms of GM- P04141 -induced HDC and P35367 expression in monocytic U937 cells were investigated . METHODS AND RESULTS : Immunohistochemistry showed that atherosclerotic human coronary and mouse ligated carotid arteries contained HDC-expressing macrophages . Gene expression of HDC , P35367 , P25021 , and GM- P04141 was also detected in the lesions . In U937 cells , GM- P04141 enhanced histamine secretion and gene expression of HDC and P35367 . A promoter assay showed that GM- P04141 enhanced gene transcription of HDC and P35367 but not P25021 . CONCLUSIONS : The present results indicate that HDC and HHR are expressed in arteriosclerotic lesion , and that GM- P04141 induces HDC and P35367 expression in monocytes . Locally produced histamine might participate in atherogenesis by affecting the expression of atherosclerosis-related genes in monocytes and smooth muscle cells . The presence of histamine-producing macrophages and gene expression of histamine receptors and GM- P04141 was demonstrated in arteriosclerotic lesions . In monocytic U937 cells , GM- P04141 upregulated the expression of histamine and P35367 . Coordinated expression of histamine and its receptors by GM- P04141 would participate in atherogenesis by affecting monocytic and SMC gene expression . Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . DB01184 treatment for gastroparesis : demographic and pharmacogenetic characterization of clinical efficacy and side-effects . BACKGROUND : DB01184 is a useful alternative to metoclopramide for treatment of gastroparesis due to better tolerability . Effectiveness and side-effects from domperidone may be influenced by patient-related factors including polymorphisms in genes encoding drug-metabolizing enzymes , drug transporters , and domperidone targets . AIMS : The aim of this study was to determine if demographic and pharmacogenetic parameters of patients receiving domperidone are associated with response to treatment or side-effects . METHODS : Patients treated with domperidone for gastroparesis provided saliva samples from which DNA was extracted . Fourteen single-nucleotide polymorphisms ( SNPs ) in seven candidate genes ( P08183 , P10635 , P14416 , P15382 , Q9Y6J6 , Q12809 , P51787 ) were used for genotyping . SNP microarrays were used to assess single-nucleotide polymorphisms in the ADRA1A , P35368 , and P25100 loci . RESULTS : Forty-eight patients treated with domperidone participated in the study . DNA was successfully obtained from each patient . Age was associated with effectiveness of domperidone ( p=0.0088 ) . Genetic polymorphism in Q12809 was associated with effectiveness of domperidone ( p=0.041 ) . The efficacious dose was associated with polymorphism in P08183 gene ( p=0.0277 ) . The side-effects of domperidone were significantly associated with the SNPs in the promoter region of P25100 gene . CONCLUSIONS : Genetic characteristics associated with response to domperidone therapy included polymorphisms in the drug transporter gene P08183 , the potassium channel Q12809 gene , and α1D -- adrenoceptor P25100 gene . Age was associated with a beneficial response to domperidone . If verified in a larger population , this information might be used to help determine which patients with gastroparesis might respond to domperidone and avoid treatment in those who might develop side-effects . Endogenous expression of histamine H1 receptors functionally coupled to phosphoinositide hydrolysis in P13671 glioma cells : regulation by cyclic AMP . 1. The effects of histamine receptor agonists and antagonists on phospholipid hydrolysis in rat-derived P13671 glioma cells have been investigated . 2 . P35367 -stimulation caused a concentration-dependent increase in the accumulation of total [ 3H ] -inositol phosphates in cells prelabelled with [ 3H ] -myo-inositol . The rank order of agonist potencies was histamine ( EC50 = 24 microM ) > N alpha-methylhistamine ( EC50 = 31 microM ) > 2-thiazolylethylamine ( EC50 = 91 microM ) . 3 . The response to 0.1 mM histamine was antagonized in a concentration-dependent manner by the H1-antagonists , mepyramine ( apparent Kd = 1 nM ) and (+)-chlorpheniramine ( apparent Kd = 4 nM ) . In addition , (-)-chlorpheniramine was more than two orders of magnitude less potent than its (+)-stereoisomer . 4 . Elevation of intracellular cyclic AMP accumulation with forskolin ( 10 microM , EC50 = 0.3 microM ) , isoprenaline ( 1 microM , EC50 = 4 nM ) or rolipram ( 0.5 mM ) , significantly reduced the histamine-mediated ( 0.1 mM ) inositol phosphate response by 37 % , 43 % and 26 % respectively . In contrast , 1,9-dideoxyforskolin did not increase cyclic AMP accumulation and had no effect on the phosphoinositide response to histamine . 5 . These data indicate the presence of functionally coupled , endogenous histamine H1 receptors in P13671 glioma cells . Furthermore , the results also indicate that H1 receptor-mediated phospholipid hydrolysis is inhibited by the elevation of cyclic AMP levels in these cells . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Pharmacological profile of astemizole-derived compounds at the histamine H1 and H4 receptor -- H1/H4 receptor selectivity . DB00637 , a P35367 antagonist shows high affinity to the histamine H1 receptor but only a moderate affinity to the histamine H4 receptor . This study aims to modify the astemizole to keep high affinity to the histamine H1 receptor and to increase affinity to the histamine H4 receptor . Therefore , 13 astemizole-derived compounds and astemizole-JNJ7777120-derived hybrid compounds were synthesized and pharmacologically characterized at the histamine H1 and H4 receptors . The new compounds show affinity to the histamine H1 receptor in the pK i range from 5.3 to 8.8 , whereas the affinity of these compounds to the histamine H4 receptor was surprisingly rather low ( pK i from 4.4 to 5.6 ) . Three representative compounds were docked into the histamine H1 receptor and molecular dynamic studies were performed to explain the binding mode and the experimental results on a molecular level . Furthermore , taking into account the binding mode of compounds with high affinity to the histamine H4 receptor , a H1/H4-pharmacophore hypothesis was developed . Different cholinesterase inhibitor effects on P04141 cholinesterases in Alzheimer patients . BACKGROUND : The current study aimed to compare the effects of different cholinesterase inhibitors on acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) activities and protein levels , in the cerebrospinal fluid ( P04141 ) of Alzheimer disease ( AD ) patients . METHODS AND FINDINGS : AD patients aged 50-85 years were randomized to open-label treatment with oral rivastigmine , donepezil or galantamine for 13 weeks . P22303 and BuChE activities were assayed by Ellman 's colorimetric method . Protein levels were assessed by enzyme-linked immunosorbent assay ( ELISA ) . Primary analyses were based on the Completer population ( randomized patients who completed Week 13 assessments ) . 63 patients were randomized to treatment . DB00989 was associated with decreased P22303 activity by 42.6 % and decreased P22303 protein levels by 9.3 % , and decreased BuChE activity by 45.6 % and decreased BuChE protein levels by 21.8 % . DB00674 decreased P22303 activity by 2.1 % and BuChE activity by 0.5 % , but increased P22303 protein levels by 51.2 % and BuChE protein levels by 10.5 % . Donepezil increased P22303 and BuChE activities by 11.8 % and 2.8 % , respectively . Donepezil caused a 215.2 % increase in P22303 and 0.4 % increase in BuChE protein levels . Changes in mean P22303 -Readthrough/Synaptic ratios , which might reflect underlying neurodegenerative processes , were 1.4 , 0.6 , and 0.4 for rivastigmine , donepezil and galantamine , respectively . CONCLUSION : The findings suggest pharmacologically-induced differences between rivastigmine , donepezil and galantamine . DB00989 provides sustained inhibition of P22303 and BuChE , while donepezil and galantamine do not inhibit BuChE and are associated with increases in P04141 P22303 protein levels . The clinical implications require evaluation . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . DB11320 Promotes the Release of P05231 via the P35367 /p38 and NF-κB Pathways in Nasal Fibroblasts . PURPOSE : Based on the close relationship between histamine and interleukin 6 ( P05231 ) , we hypothesized that histamine may regulate the production of cytokines , such as P05231 , during allergic inflammation . Here , we examined the role of histamine in P05231 production and histamine receptor activity in nasal fibroblasts , along with the mechanisms underlying these effects . METHODS : Experiments were performed using nasal fibroblasts from 8 normal patients . RT-PCR was used to identify the major histamine receptors expressed in nasal fibroblasts . Fibroblasts were then treated with histamine with or without histamine-receptor antagonists , and monitored for P05231 production using an ELISA . Four potential downstream signaling molecules , p38 , extracellular signal-regulated kinase ( P29323 ) , c-Jun N-terminal kinase ( JNK ) , and NF-κB , were evaluated by Western blot , and a luciferase reporter assay . RESULTS : Elevated expression was seen for all histamine receptors , with P05231 protein levels increasing significantly following histamine stimulation . Among the histamine-receptor specific antagonists , only the P35367 antagonist significantly decreased P05231 production in histamine-stimulated nasal fibroblasts . DB11320 increased the expression level of phosphorylated p38 ( pp38 ) , pERK , and pJNK , as well as NF-κB induction . The P35367 antagonist actively suppressed pp38 and NF-κB expression in histamine-induced nasal fibroblasts , but not pERK and pJNK . The p38 inhibitor strongly attenuated P05231 production in histamine-stimulated nasal fibroblasts . CONCLUSIONS : The data presented here suggest that antihistamines may be involved in the regulation of cytokines , such as P05231 , due to the role of histamine as an inflammatory mediator in nasal fibroblasts . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively .

MH_train_58

MH_train_58

MH_train_58

interacts_with DB01367?

multiple_choice

Q16552 stimulates the production of inflammatory mediators via Erk1/2 , p38 MAPK , PI3K/Akt , and NF-κB pathways in ARPE-19 cells . PURPOSE : To investigate the signaling pathways involved in interleukin ( IL ) -17A -mediated production of interleukin 8 ( P10145 ) , chemokine ( C-C motif ) ligand 2 ( P13500 ) , and interleukin 6 ( P05231 ) by ARPE-19 cells , a spontaneously arisen cell line of retinal pigment epithelium ( Q96AT9 ) . METHODS : Flow cytometry analysis and western blot were used to detect the phosphorylation of extracellular signal-regulated kinases 1/2 ( Erk1/2 ) , p38 mitogen activated protein kinase ( MAPK ) and protein kinase B ( P31749 ; Akt ) in ARPE-19 cells stimulated with Q16552 . These cells were further pretreated with a series of kinase inhibitors and followed by incubation with Q16552 . P10145 , P13500 , and P05231 in the supernatant were quantified by enzyme-linked immunosorbent assay ( ELISA ) . RESULTS : Coculture of ARPE-19 cells with Q16552 resulted in significant increases in Erk1/2 , p38 MAPK , and Akt phosphorylation . Inhibition of p38MAPK , phosphoinositide 3-kinase ( PI3K ) -Akt and nuclear factor-kappaB ( NF-κB ) , with the inhibitors SB203580 , LY294002 and pyrrolydine dithiocarbamate ( PDTC ) respectively , reduced Q16552 ( 100 ng/ml ) mediated production of P10145 , P13500 , and P05231 in a concentration dependent manner . Inhibition of Erk1/2 with PD98059 decreased the expression of the tested three inflammatory mediators when using low doses of Q16552 ( 0-10 ng/ml ) but not at higher concentrations . CONCLUSIONS : Q16552 -induced production of inflammatory mediators by ARPE-19 cells involves Erk1/2 , p38MAPK , PI3K-Akt and NF-κB pathways . Molecular pathways : the basis for rational combination using MEK inhibitors in P01116 -mutant cancers . Mutations in DB01367 oncogenes are frequently observed in human cancers , and the mutations result in activation of the DB01367 -RAF-MEK- P29323 pathway , leading to cell proliferation and survival . The pathway is , therefore , a potent therapeutic target in the DB01367 -mutant cancers . MEK inhibitors can specifically block the pathway and are one of the key types of drugs for the treatment of the DB01367 -mutant cancers . As DB01367 proteins activate other downstream signaling proteins in addition to the DB01367 -RAF-MEK- P29323 pathway , combination therapeutic approaches with MEK inhibitors are also being evaluated . Moreover , MEK inhibitors can arrest cancer cells in P55008 phase and repress prosurvival Bcl2 family proteins such as Q07820 and P10415 /BCLXL , and increase expression of Bim , a proapoptotic BH3-only family protein . This mechanism may explain the efficacy of the combination of MEK inhibitors with cytotoxic agents or other targeted inhibitors . A better understanding of the pathway will help us with development of rational combinations for the treatment of the DB01367 -mutant cancers . [ DB00988 -beta-hydroxylaseaktivität im Plasma von Dialysepatienten ( author 's transl ) ] . Plasma dopamin-b-hydroxylase ( P09172 ) was studied in 70 healthy control persons and in 37 hemodialysed patients . Basal P09172 in controls corresponded to 50.0 +/- 29.3 IU . There was was no significant difference between males ( 53.9 +/1 33.8 IU ) and females ( 47.4 +/- 25 IU ) ; no correlation could be found between age and plasma P09172 . In hemodialysed patients basal P09172 levels were significantly ( p less than 0.01 ) decreased ( 32.5 % /- 17.6 IU ) , suggesting lowered sympathetic activity and/or abnormalities in release , distribution space , or metabolism of P09172 . During hemodialysis plasma P09172 activity rose during ultrafiltration . This finding indicates a directionally appropriate sympathetic reflex response to volume depletion in dialysed patients . Candidate genes for the hypoxic tumor phenotype . In this study , we have analyzed changes induced by hypoxia at the transcriptional level of genes that could be responsible for a more aggressive phenotype . Using a series of DNA array membranes , we identified a group of hypoxia-induced genes that included plasminogen activator inhibitor-1 ( P05121 ) , insulin-like growth factor-binding protein 3 ( P17936 ) , endothelin-2 , low-density lipoprotein receptor-related protein ( Q14764 ) , P10415 -interacting killer ( Q13323 ) , migration-inhibitory factor ( MIF ) , matrix metalloproteinase-13 ( P45452 ) , fibroblast growth factor-3 ( P11487 ) , P24522 , and vascular endothelial growth factor ( P15692 ) . The induction of each gene was confirmed by Northern blot analysis in two different squamous cell carcinoma-derived cell lines . We also analyzed the kinetics of P05121 induction by hypoxia in more detail because it is a secreted protein that may serve as a useful molecular marker of hypoxia . On exposure to hypoxia , there was a gradual increase in P05121 mRNA between 2 and 24 h of hypoxia followed by a rapid decay after 2 h of reoxygenation . P05121 levels were also measured in the serum of a small group of head and neck cancer patients and were found to correlate with the degree of tumor hypoxia found in these patients . G protein-coupled receptor kinase-3-deficient mice exhibit WHIM syndrome features and attenuated inflammatory responses . Chemokine receptor interactions coordinate leukocyte migration in inflammation . Chemokine receptors are GPCRs that when activated , are phosphorylated by GRKs to turn off G protein-mediated signaling yet recruit additional signaling machinery . Recently , P35626 was identified as a negative regulator of P48061 / P61073 signaling that is defective in human WHIM syndrome . Here , we report that P35626 -/- mice exhibit numerous features of human WHIM , such as impaired P48061 -mediated desensitization , enhanced P61073 signaling to P29323 activation , altered granulocyte migration , and a mild myelokathexis . Moreover , P35626 -/- protects mice from two acute models of inflammatory arthritis ( K/BxN serum transfer and CAIA ) . In these granulocyte-dependent disease models , protection of P35626 -/- mice is mediated by retention of cells in the marrow , fewer circulating granulocytes in the peripheral blood , and reduced granulocytes in the joints during active inflammation . In contrast to WHIM , P35626 -/- mice have minimal hypogammaglobulinemia and a peripheral leukocytosis with increased lymphocytes and absent neutropenia . Thus , we conclude that the loss of P35626 -mediated regulation of P48061 / P61073 signaling contributes to some , but not all , of the complete WHIM phenotype and that P35626 inhibition may be beneficial in the treatment of inflammatory arthritis . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal . A surface plasmon resonance-based solution affinity assay for heparan sulfate-binding proteins . A surface plasmon resonance-based solution affinity assay is described for measuring the K(d) of binding of heparin/heparan sulfate-binding proteins with a variety of ligands . The assay involves the passage of a pre-equilibrated solution of protein and ligand over a sensor chip onto which heparin has been immobilised . DB01109 sensor chips prepared by four different methods , including biotin-streptavidin affinity capture and direct covalent attachment to the chip surface , were successfully used in the assay and gave similar K(d) values . The assay is applicable to a wide variety of heparin/HS-binding proteins of diverse structure and function ( e.g. , P05230 , P09038 , P15692 , P10145 , P80075 , P01008 , P02776 ) and to ligands of varying molecular weight and degree of sulfation ( e.g. , heparin , DB05808 , sucrose octasulfate , naphthalene trisulfonate ) and is thus well suited for the rapid screening of ligands in drug discovery applications . Genetic pathway-based hierarchical clustering analysis of older adults with cognitive complaints and amnestic mild cognitive impairment using clinical and neuroimaging phenotypes . Hierarchical clustering is frequently used for grouping results in expression or haplotype analyses . These methods can elucidate patterns between measures that can then be applied to discerning their validity in discriminating between experimental conditions . Here a hierarchical clustering method is used to analyze the results of an imaging genetics study using multiple brain morphology and cognitive testing endpoints for older adults with amnestic mild cognitive impairment ( D6RGH6 ) or cognitive complaints ( CC ) compared to healthy controls ( HC ) . The single nucleotide polymorphisms ( SNPs ) are a subset of those included on a larger array that are found in a reported Alzheimer 's disease ( AD ) and neurodegeneration pathway . The results indicate that genetic models within the endpoints cluster together , while there are 4 distinct sets of SNPs that differentiate between the endpoints , with most significant results associated with morphology endpoints rather than cognitive testing of patients ' reported symptoms . The genes found in at least one cluster are P08183 , Q02410 , P56817 , Q9Y5Z0 , P10415 , Q07817 , P55210 , P28329 , P01034 , P35462 , P21918 , P05231 , Q07954 , NAT1 , and P49810 . The greater associations with morphology endpoints suggests that changes in brain structure can be influenced by an individual 's genetic background in the absence of dementia and in some cases ( Cognitive Complaints group ) even without those effects necessarily being detectable on commonly used clinical tests of cognition . The results are consistent with polygenic influences on early neurodegenerative changes and demonstrate the effectiveness of hierarchical clustering in identifying genetic associations among multiple related phenotypic endpoints . Targeting P00441 reduces experimental non–small-cell lung cancer . Approximately 85 % of lung cancers are non–small-cell lung cancers ( NSCLCs ) , which are often diagnosed at an advanced stage and associated with poor prognosis . Currently , there are very few therapies available for NSCLCs due to the recalcitrant nature of this cancer . Mutations that activate the small GTPase P01116 are found in 20 % to 30 % of NSCLCs . Here , we report that inhibition of superoxide dismutase 1 ( P00441 ) by the small molecule DB05088 induced cell death in various NSCLC cells , including those harboring P01116 mutations . ATN-224–dependent P00441 inhibition increased superoxide , which diminished enzyme activity of the antioxidant glutathione peroxidase , leading to an increase in intracellular hydrogen peroxide ( H(2)O(2) ) levels . We found that ATN-224–induced cell death was mediated through H(2)O(2)-dependent activation of O75791 MAPK and that O75791 activation led to a decrease in the antiapoptotic factor Q07820 , which is often upregulated in NSCLC . Treatment with both DB05088 and DB05764 , an inhibitor of the apoptosis regulators P10415 /BCLXL , augmented cell death . Furthermore , we demonstrate that DB05088 reduced tumor burden in a mouse model of NSCLC . Our results indicate that antioxidant inhibition by DB05088 has potential clinical applications as a single agent , or in combination with other drugs , for the treatment of patients with various forms of NSCLC , including P01116 -driven cancers . Alternative inclusion of fibroblast growth factor receptor 2 exon IIIc in Dunning prostate tumors reveals unexpected epithelial mesenchymal plasticity . In epithelial cells , alternative splicing of fibroblast growth factor receptor 2 ( P21802 ) transcripts leads to the expression of the P21802 (IIIb) isoform , whereas in mesenchymal cells , the same process results in the synthesis of P21802 (IIIc) . Expression of the P21802 (IIIc) isoform during prostate tumor progression suggests a disruption of the epithelial character of these tumors . To visualize the use of P21802 exon IIIc in prostate P01008 tumors in syngeneic rats , we constructed minigene constructs that report on alternative splicing . Imaging these alternative splicing decisions revealed unexpected mesenchymal-epithelial transitions in these primary tumors . These transitions were observed more frequently where tumor cells were in contact with stroma . Indeed , these transitions were frequently observed among lung micrometastases in the organ parenchyma and immediately adjacent to blood vessels . Our data suggest an unforeseen relationship between epithelial mesenchymal plasticity and malignant fitness . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Simultaneous inhibition of MEK and P11802 leads to potent apoptosis in human melanoma cells . ABSTRACT Deregulation of DB01367 -RAF-MEK- P29323 and P42771 -cycylin D: P11802 /6-RB pathways is important for melanoma development . Chemotherapeutic agents targeting both pathways were developed but results of clinical studies with monotherapies were disappointing . We examined the effect of co-targeting both pathways with MEK inhibitor PD98059 and P11802 inhibitor 219476 on human melanoma cells lines , and found that combinatorial treatment dramatically increased apoptosis compared to the single agent treatment . The apoptosis was associated with downregulation of P10415 , Q07817 , O15392 , and upregulation of O43521 . Our results indicate that simultaneously targeting P29323 and RB pathways is a promising strategy for melanoma treatment and should encourage further in-depth investigations . Altered decorin leads to disrupted endothelial cell function : a possible mechanism in the pathogenesis of fetal growth restriction ? OBJECTIVE : Fetal growth restriction ( P09769 ) is a key cause of adverse pregnancy outcome where maternal and fetal factors are identified as contributing to this condition . Idiopathic P09769 is associated with altered vascular endothelial cell functions . P07585 ( P07585 ) has important roles in the regulation of endothelial cell functions in vascular environments . P07585 expression is reduced in P09769 . The objectives were to determine the functional consequences of reduced P07585 in a human microvascular endothelial cell line model ( HMVEC ) , and to determine downstream targets of P07585 and their expression in primary placental microvascular endothelial cells ( PLECs ) from control and P09769 -affected placentae . APPROACH : Short-interference RNA was used to reduce P07585 expression in HMVECs and the effect on proliferation , angiogenesis and thrombin generation was determined . A Growth Factor PCR Array was used to identify downstream targets of P07585 . The expression of target genes in control and P09769 PLECs was performed . RESULTS : P07585 reduction decreased proliferation and angiogenesis but increased thrombin generation with no effect on apoptosis . The array identified three targets of P07585 : O60258 , Q14116 and O14793 . Validation of target genes confirmed decreased expression of P15692 , P14780 , EGFR1 , P12314 and P49763 in HMVECs and PLECs from control and P09769 pregnancies . CONCLUSIONS : Reduction of P07585 in vascular endothelial cells leads to disrupted cell functions . The targets of P07585 include genes that play important roles in angiogenesis and cellular growth . Therefore , differential expression of these may contribute to the pathogenesis of P09769 and disease states in other microvascular circulations . Unbiased screen for interactors of leucine-rich repeat kinase 2 supports a common pathway for sporadic and familial Parkinson disease . Mutations in leucine-rich repeat kinase 2 ( Q5S007 ) cause inherited Parkinson disease ( PD ) , and common variants around Q5S007 are a risk factor for sporadic PD . Using protein-protein interaction arrays , we identified P10415 -associated athanogene 5 , Rab7L1 ( P51149 , member DB01367 oncogene family-like 1 ) , and O14976 as binding partners of Q5S007 . The latter two genes are candidate genes for risk for sporadic PD identified by genome-wide association studies . These proteins form a complex that promotes clearance of Golgi-derived vesicles through the autophagy-lysosome system both in vitro and in vivo . We propose that three different genes for PD have a common biological function . More generally , data integration from multiple unbiased screens can provide insight into human disease mechanisms . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . Phorbol ester-mediated inhibition of growth and regulation of proto-oncogene expression in the human T cell leukemia line JURKAT . The expression and function of several proto-oncogenes were examined in a human acute T cell leukemia line , JURKAT , during phorbol ester-induced terminal differentiation . Treating JURKAT cells with the phorbol ester tetradecanoyl phorbol acetate ( TPA ) inhibited their proliferation and induced expression of the gene for the interleukin 2 receptor alpha chain ( IL2R-alpha ) , consistent with previous reports . In unstimulated proliferating JURKAT cells , high levels of C-MYC , N- DB01367 , and P10415 mRNAs were found that diminished rapidly following TPA-induced cessation of growth . In contrast , accumulation of mRNAs for the C- P01100 , C- P05412 , and P18146 genes increased markedly in TPA-treated cells and preceded the induction of IL2R-alpha mRNA . Expression of C- P10242 , C-RAF-1 , C- P06239 , C- P06241 , and C- P09769 proto-oncogenes was relatively unchanged . To explore directly the function of two of these protooncogenes in regulating the growth of JURKAT T cells , we stably transferred C-MYC and P10415 expression plasmids into these cells . Despite sustained expression of C-MYC , P10415 , or the combination of these protooncogenes , TPA continued to inhibit JURKAT cell growth and to induce IL2R expression . Thus , although C-MYC and P10415 proto-oncogene expression correlated with proliferation in TPA-treated JURKAT cells , continuous over-expression of even the combination of these oncogenes was insufficient for abrogating the effects of TPA in these leukemic T cells . Because human lymphoid malignancies frequently contain chromosomal translocations that deregulate the expression of C-MYC and P10415 , our findings could have relevance for attempts to induce terminal differentiation of leukemic cells by in vitro exposure of patients ' bone marrow cells to phorbol esters . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . P09455 -1 expression in ovarian cancer : a potential therapeutic target . BACKGROUND/AIM : Cellular retinol binding protein-1 regulates retinol bioavailability and contributes to cell differentiation maintenance , but its role in ovarian carcinogenesis remains uncertain . We investigated P09455 -1 expression in ovarian tumors and P09455 -1 signaling-regulated pathways . MATERIALS AND METHODS : We performed immunohistochemistry , methylation-specific PCR , gene copy number analysis in ovarian tumors and proliferation/apoptosis evaluation , gene array , blot and real-time PCR in P09455 -1-transfected A2780 ovarian cancer cells . RESULTS : P09455 -1 expression was reduced or absent in G2 and P46379 ovarian carcinomas . P09455 -1 silencing in 60 % of G2 and 66.7 % of P46379 carcinomas was due to P09455 -1 promoter methylation . A2780 P09455 -1-transfected cells showed increased retinol-induced apoptosis , retinoid-induced reduced clonogenicity and down-regulation of proliferation and transcription genes , including P31749 , Q9Y243 , P00533 , P01100 , P05412 , P42224 and P42229 . CONCLUSION : P09455 -1 loss in G2/ P46379 ovarian carcinomas and increased apoptotic susceptibility to retinoids in P09455 -1-transfected-A2780 cells suggest P09455 -1 screening as a target to ensure efficacy of an adjuvant retinoid therapy . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Genetic and epigenetic markers in the evaluation of pancreatic masses . BACKGROUND : Methylation markers have shown promise in the early diagnosis of pancreatic carcinoma . The aim of this study was to assess the diagnostic utility of hypermethylation status of candidate genes in combination with P01116 mutation detection in the evaluation of pancreatic masses . EXPERIMENTAL DESIGN : Sixty-one fine needle aspirates of pancreatic masses ( 43 pancreatic adenocarcinomas and 18 chronic pancreatitis ) were studied . Methylation status of P25021 , Q05925 , P09486 , P55290 and P25054 were analysed using melting curve analysis after DNA bisulfite treatment . P01116 mutations were also analysed . RESULTS : The methylation panel had a sensitivity of 73 % ( 27 of 37 , CI 95 % 56 to 86 % ) and a specificity of 100 % whenever two or more promoters were found hypermethylated . P01116 mutations showed a sensitivity of 77 % ( 33 of 43 , CI 95 % 62 to 88 % ) and a specificity of 100 % . Both molecular analyses added useful information to cytology by increasing the number of informative cases . When genetic and epigenetic analyses were combined sensitivity was 84 % ( 36 of 43 CI 95 % 69 to 93 % ) maintaining a 100 % specificity . CONCLUSIONS : Analysis of hypermethylation status of a panel of genes and P01116 mutation detection offer a similar diagnostic yield in the evaluation of pancreatic masses . The combined molecular analysis increases the number of informative cases without diminishing specificity . Linkage analysis of multiple sclerosis with candidate region markers in Sardinian and Continental Italian families . Previous genome screens in multiple sclerosis have shown some evidence of linkage in scattered chromosomal regions . Although in no case the evidence of each single study was compelling and although in general the linkage ' peaks ' of the different studies did not coincide , some regions can be considered likely candidates for the presence of MS risk genes because of the clustering of MLS scores and homology with eae loci . We performed a linkage analysis of markers in these regions and of intragenic markers of some individual candidate genes ( HLA- Q8IUH3 , P16410 , P15248 , P02649 , P10415 , P20333 ) . For the first time , Southern European populations were targeted , namely Continental Italians and Sardinians . A total of 69 multiplex families were typed for 67 markers by a semi-automatic fluorescence-based assay . Results were analysed for linkage by two non-parametric tests : GENEHUNTER and SimIBD . In general , the linkage scores obtained were low , confirming the conclusion that no gene is playing a major role in the disease . However , some markers , in 2p11 , 3q21.1 , 7p15.2 and 22q13.1 stood out as promising since they showed higher scores with one or the other test . This stimulates further association analysis of a large number of simplex families from the same populations . Reactive oxygen species , DNA damage , and error-prone repair : a model for genomic instability with progression in myeloid leukemia ? Myelodysplastic syndromes ( P43034 ) comprise a heterogeneous group of disorders characterized by ineffective hematopoiesis , with an increased propensity to develop acute myelogenous leukemia ( AML ) . The molecular basis for P43034 progression is unknown , but a key element in P43034 disease progression is loss of chromosomal material ( genomic instability ) . Using our two-step mouse model for myeloid leukemic disease progression involving overexpression of human mutant P01111 and P10415 genes , we show that there is a stepwise increase in the frequency of DNA damage leading to an increased frequency of error-prone repair of double-strand breaks ( DSB ) by nonhomologous end-joining . There is a concomitant increase in reactive oxygen species ( ROS ) in these transgenic mice with disease progression . Importantly , P63000 , an essential component of the ROS-producing NADPH oxidase , is downstream of DB01367 , and we show that ROS production in P01111 / P10415 mice is in part dependent on P63000 activity . DNA damage and error-prone repair can be decreased or reversed in vivo by N-acetyl cysteine antioxidant treatment . Our data link gene abnormalities to constitutive DNA damage and increased DSB repair errors in vivo and provide a mechanism for an increase in the error rate of DNA repair with P43034 disease progression . These data suggest treatment strategies that target DB01367 / P31749 pathways and ROS production in human P43034 /AML . P60484 gene silencing prevents HIV-1 gp120(IIIB)-induced degeneration of striatal neurons . To assess the role of the phosphatase and tensin homologue on chromosome 10 ( P60484 ) in mediating envelope glycoprotein 120 (gp120)-induced neurotoxicity in the striatum , P60484 was silenced using short interfering RNA ( siRNA ) vectors . P60484 activity directs multiple downstream pathways implicated in gp120-induced neuronal injury and death . P60484 is a negative regulator of Akt ( protein kinase B ) phosphorylation , but has also been shown to directly activate extrasynaptic DB01221 receptors and dephosphorylate focal adhesion kinase . Rodent striatal neurons were nucleofected with green fluorescent protein ( GFP ) -expressing siRNA constructs to silence P60484 ( PTENsi-GFP ) or with negative-control ( NCsi-GFP ) vectors , and exposed to HIV-1 gp120(IIIB) using rigorously controlled , cell culture conditions including computerized time-lapse microscopy to track the fate of individual neurons following gp120 exposure . Immunofluorescence labeling showed that subpopulations of striatal neurons possess P61073 and P51681 co-receptor immunoreactivity and that gp120(IIIB) was intrinsically neurotoxic to isolated striatal neurons . Importantly , PTENsi-GFP , but not control NCsi-GFP , constructs markedly decreased P60484 mRNA and protein levels and significantly attenuated gp120-induced death . These findings implicate P60484 as a critical factor in mediating the direct neurotoxic effects of HIV-1 gp120 , and suggest that effectors downstream of P60484 such as Akt or other targets are potentially affected . The selective abatement of P60484 activity in neurons may represent a potential therapeutic strategy for the CNS complications of HIV-1 . Complementation by P10415 and C-HA- DB01367 oncogenes in malignant transformation of rat embryo fibroblasts . The P10415 ( B cell lymphoma/leukemia-2 ) and C-HA- DB01367 oncogenes encode membrane-associated proteins of 26 and 21 kilodaltons , respectively . Although DB01367 proteins have long been known for their ability to bind and hydrolyze GTP , recent investigations suggest that P10415 encodes a novel GTP-binding protein ( S. Haldar , C. Beatty , Y. Tsujimoto , and C. M. Croce , Nature [ London ] 342:195-198 , 1989 ) . Cotransfection of P10415 and HA- DB01367 oncogenes resulted in morphological transformation of early-passage rodent fibroblasts , rendering these cells tumorigenic in animals and enabling them to grow in semisolid medium . In contrast , cotransfection of P10415 with oncogenes that encode nuclear proteins ( E1A and C-MYC ) did not produce malignant transformation , whereas HA- DB01367 did complement with these genes . These findings suggest that proteins encoded by oncogenes such as P10415 and HA- DB01367 , although having similar subcellular locations and perhaps similar biochemical properties , can regulate distinct complementary pathways involved in cellular transformation . A new platinum complex of triazine demonstrates P55008 arrest with novel biological profile in human breast cancer cell line , MDA-MB-468 . A novel class of platinum(II) complexes of pyridine sulfide derivatives of triazine was synthesized , characterized , and investigated using the human breast cancer cell line , MDA-MB-468 . S-30 was one of the most potent derivatives of its class ( IC(50) , 0.39 microM ) eliciting the greatest biological response . S-30 induced arrest in the P55008 phase and apoptosis ( TUNEL assay ) in a p53/ P38936 ( P38936 /CIP1)-consistent manner . Modeling and docking experiments were performed for three known targets for cisplatin , d(GpG) , d(ApG) , and a protein ( Cu/Zn superoxide dismutase , SOD ) from bovine origin . A Blast search of bovine SOD was performed to identify analogous human protein targets resulting in about 22 human proteins . A multi-sequence alignment of those targets showed > 80 % sequence identity and > 88 % similarity . One of them is P00441 that is differentially expressed ( based on global gene expression pattern ) in various forms of cancer and other diseases . P00441 controls apoptosis via p53/ Q92934 / Q07812 / P10415 in the amyotrophic lateral sclerosis ( P35858 ) pathway and is also involved in various other KEGG 's pathways . Results suggest that the S-30 is a potential cytotoxic agent . Genetic basis of delay discounting in frequent gamblers : examination of a priori candidates and exploration of a panel of dopamine-related loci . INTRODUCTION : Delay discounting is a behavioral economic index of impulsivity that reflects preferences for small immediate rewards relative to larger delayed rewards . It has been consistently linked to pathological gambling and other forms of addictive behavior , and has been proposed to be a behavioral characteristic that may link genetic variation and risk of developing addictive disorders ( i.e. , an endophenotype ) . Studies to date have revealed significant associations with polymorphisms associated with dopamine neurotransmission . The current study examined associations between delay discounting and both previously linked variants and a novel panel of dopamine-related variants in a sample of frequent gamblers . METHODS : Participants were 175 weekly gamblers of European ancestry who completed the Monetary Choice Questionnaire to assess delay discounting preferences and provided a DNA via saliva . RESULTS : In a priori tests , two loci previously associated with delayed reward discounting ( rs1800497 and rs4680 ) were not replicated , however , the long form of P21917 VNTR was significantly associated with lower discounting of delayed rewards . Exploratory analysis of the dopamine-related panel revealed 11 additional significant associations in genes associated with dopamine synthesis , breakdown , reuptake , and receptor function ( P35462 , Q01959 , DDC , P09172 , and Q05940 ) . An aggregate genetic risk score from the nominally significant loci accounted for 17 % of the variance in discounting . Mediational analyses largely supported the presence of indirect effects between the associated loci , delay discounting , and pathological gambling severity . CONCLUSIONS : These findings do not replicate previously reported associations but identify several novel candidates and provide preliminary support for a systems biology approach to understand the genetic basis of delay discounting . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Traps to catch unwary oncogenes . The MYC proto-oncogene has long been implicated in the control of normal cell growth and its deregulation is associated with the development of neoplasia . The MYC protein has a well-established role as a component of signal-transduction pathways promoting both proliferation and apoptosis . Because signalling pathways that drive cell death and cell proliferation are so tightly coupled , a synergy between genetic lesions leading to suppression of cell death and those promoting cell proliferation is observed during carcinogenesis . We discuss such synergy with respect to the cooperating oncogenes MYC , DB01367 and P10415 . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . NSA9 , a human prothrombin kringle-2-derived peptide , acts as an inhibitor of kringle-2-induced activation in EOC2 microglia . In neurodegenerative diseases , such as Alzheimer 's and Parkinson 's , microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds . P00734 and kringle-2 increase levels of NO and the mRNA expression of P35228 , IL-1beta , and P01375 in microglial cells . In contrast , the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1beta , P01375 , and P05231 in LPS-activated EOC2 microglia . In this study , we investigated the anti-inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia . Treatment with 20-100 muM of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation . NO production induced by MAPKs and NF-kappaB was similarly reduced by inhibitors of P29323 ( PD98059 ) , p38 ( SB203580 ) , NF-kappaB ( DB06151 ) , and NSA9 . These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2 . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Induction of apoptosis by a dominant negative H- DB01367 mutant ( 116Y ) in K562 cells . Recent extensive work on apoptosis has begun to reveal its molecular mechanisms . Several genes that regulate apoptosis have been identified . Among them , the P10415 gene is considered to be an important gene that inhibits apoptosis . However , there must be other genes , yet to be identified , which suppress apoptosis . It has been suggested that the activation of DB01367 function by P11274 - P00519 fusion protein in chronic myelogenous leukemia may be an important mechanism in the P11274 - P00519 mediated transformation . Therefore , in this study we have investigated whether the suppression of endogenous H- DB01367 function inhibits the P11274 - P00519 mediated transforming activity in a K562 human chronic myelogenous leukemia cell line . The induced expression of a dominant negative v-H- DB01367 mutant ( 116Y ) in K562 cells has resulted in cell death . The morphological characteristics and the detection of fragmented DNA by gel electrophoresis in the dead cells have revealed that this cell death is apoptosis . These results directly indicate that the DB01367 gene as well as the P10415 gene has an ability to suppress apoptosis . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . Follicular lymphoma cell lines , an in vitro model for antigenic selection and cytokine-mediated growth regulation of germinal centre B cells . In the periphery , B cells differentiate in germinal centres ( GCs ) of secondary lymphoid organs . Isolated GC cells die quickly in vitro by apoptosis . Therefore , cell lines originating from follicular lymphomas , which are the malignant counterparts of GC B cells , would provide a stable in vitro model to study the immunobiology of GC B cells . We have established three novel human follicular lymphoma cell lines that were characterized with special reference to immunophenotypic features , response to B-cell receptor ( P11274 ) triggering , response to cytokines and cytokine mRNA expression . One of the cell lines , HF-1A3 , has a phenotype of a centrocyte . It expresses surface immunoglobulin G ( sIgG ) and dies by apoptosis following P11274 cross-linking . Co-stimulation with interleukin-6 ( P05231 ) , P40933 or interferon-gamma ( P01579 ) rescues HF-1A3 cells from P11274 -induced apoptosis . The second cell line , HF-28 , also represents phenotypically an IgG+ centrocyte . Ligation of its P11274 leads to the cell-cycle arrest at P55008 instead of apoptosis . HF-28 cells express both CD45RA and RO isoforms , which is unusual in B lymphocytes apart from plasma cells , thus suggesting a transition to plasma cell phenotype . The third cell line , HF-4.9 , which phenotypically represents an sIgM+ centroblast , responds by proliferation to P11274 cross-linking . These cell lines offer a unique in vitro model to study antigenic selection and cytokine-mediated growth regulation of human GC B cells . Risk stratification of intermediate-risk acute myeloid leukemia : integrative analysis of a multitude of gene mutation and gene expression markers . Numerous molecular markers have been recently discovered as potential prognostic factors in acute myeloid leukemia ( AML ) . It has become of critical importance to thoroughly evaluate their interrelationships and relative prognostic importance . Gene expression profiling was conducted in a well-characterized cohort of 439 AML patients ( age < 60 years ) to determine expression levels of EVI1 , P19544 , P10415 , P08183 , Q8WXS3 , P36888 , P28906 , P14902 , ERG and Q10571 . A variety of AML-specific mutations were evaluated , that is , P36888 , P06748 , N- DB01367 , K- DB01367 , O75874 , P48735 , and P49715 (DM/SM) ( double/single ) . Univariable survival analysis shows that ( 1 ) patients with P36888 (ITD) mutations have inferior overall survival ( OS ) and event-free survival ( O43281 ) , whereas P49715 (DM) and P06748 mutations indicate favorable OS and O43281 in intermediate-risk AML , and ( 2 ) high transcript levels of Q8WXS3 , P28906 , Q10571 , EVl1 , and ERG predict inferior OS and O43281 . In multivariable survival analysis , P28906 , ERG , and P49715 (DM) remain significant . Using survival tree and regression methodologies , we show that P49715 (DM) , P28906 , and P48735 mutations are capable of separating the intermediate group into 2 AML subgroups with highly distinctive survival characteristics ( OS at 60 months : 51.9 % vs 14.9 % ) . The integrated statistical approach demonstrates that from the multitude of biomarkers a greatly condensed subset can be selected for improved stratification of intermediate-risk AML . Central role for hydrogen peroxide in P47900 ADP receptor-mediated cellular responses in vascular endothelium . ADP activates a family of cell surface receptors that modulate signaling pathways in a broad range of cells . ADP receptor antagonists are widely used to treat cardiovascular disease states . These studies identify a critical role for the stable reactive oxygen species hydrogen peroxide ( H2O2 ) in mediating cellular responses activated by the G protein-coupled P47900 receptor for ADP . We found that ADP-dependent phosphorylation of key endothelial signaling proteins -- including endothelial nitric oxide synthase , AMP-activated protein kinase , and the actin-binding P29966 protein -- was blocked by preincubation with PEG-catalase , which degrades H2O2 . ADP treatment promoted the H2O2-dependent phosphorylation of c-Abl , a nonreceptor tyrosine kinase that modulates the actin cytoskeleton . Cellular imaging experiments using fluorescence resonance energy transfer-based biosensors revealed that ADP-stimulated activation of the cytoskeleton-associated small GTPase Rac1 was independent of H2O2 . However , Rac1-dependent activation of AMP-activated protein kinase , the signaling phospholipid phosphatidylinositol- ( 4 , 5 ) -bisphosphate , and the c-Abl-interacting protein CrkII are mediated by H2O2 . We transfected endothelial cells with differentially targeted HyPer2 H2O2 biosensors and found that ADP promoted a marked increase in H2O2 levels in the cytosol and caveolae , and a smaller increase in mitochondria . We performed a screen for P47900 receptor-mediated receptor tyrosine kinase transactivation and discovered that ADP transactivates P36888 ( Flt3 ) , a receptor tyrosine kinase expressed in these cells . Our observation that P47900 receptor-mediated responses involve Flt3 transactivation may identify a unique mechanism whereby cancer chemotherapy with receptor tyrosine kinase inhibitors promotes vascular dysfunction . Taken together , these findings establish a critical role for endogenous H2O2 in control of ADP-mediated signaling responses in the vascular wall . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . cDNA array analysis of cytobrush-collected normal and malignant cervical epithelial cells : a feasibility study . Analysis of gene expression pattern is a useful approach to evaluating the biological behavior and clinical outcome of several human malignancies . Differentially expressed genes in malignant squamous cervical cells and the feasibility of gene expression profiling on squamous cervical cells obtained from cervical swabs were investigated . Cervical squamous cells from three women with high-risk human papilloma virus ( HR-HPV ) positive invasive squamous cervical carcinoma and from three HPV-negative women with normal ectocervical smears were analyzed with cDNA array . Immunoblot analysis was performed to detect the proteins corresponding to the highest upregulated genes with cDNA array . mRNA expression of P04626 , P10721 , P17948 , P04198 , DB01367 , CDKN2A , P24385 , P15531 , P22392 , MET , P21781 , P21802 , and P42224 was increased in malignant samples . Several expressed genes associated with antiapoptosis ( such as P10415 ) , cell structuring , or cell attachment were also upregulated in carcinoma cells . Decreased gene expression was observed for members of the transforming growth factor receptor superfamily ( TGF ) and integrin family , interleukin 1 ( IL1 ) , and insulin-like growth factor binding proteins ( IGFBPs ) . This study shows the feasibility of gene expression profiling of cervical squamous cells obtained with cytobrushes by identifying a characteristic gene expression pattern that clearly distinguishes between malignant and normal cervical epithelia of squamous type . We hypothesize that this noninvasive technique could be used in the evaluation of ambiguous Papanicolaou ( PAP ) smears . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Aberrant microRNA expression likely controls DB01367 oncogene activation during malignant transformation of human prostate epithelial and stem cells by arsenic . Inorganic arsenic ( iAs ) , a human carcinogen , potentially targets the prostate . iAs malignantly transforms the RWPE-1 human prostate epithelial line to CAsE-PE cells , and a derivative normal stem cell ( SC ) line , WPE-stem , to As-Cancer SC ( As-CSC ) line . MicroRNAs ( miRNA ) are noncoding but exert negative control on expression by degradation or translational repression of target mRNAs . Aberrant miRNA expression is important in carcinogenesis . A miRNA array of CAsE-PE and As-CSC revealed common altered expression in both for pathways concerning oncogenesis , miRNA biogenesis , cell signaling , proliferation , and tumor metastasis and invasion . The P01116 oncogene is overexpressed in CAsE-PE cells but not by mutation or promoter hypomethylation , and is intensely overexpressed in As-CSC cells . In both transformants , decreased miRNAs targeting P01116 and DB01367 superfamily members occurred . Reduced miR-134 , miR-373 , miR-155 , miR-138 , miR-205 , miR-181d , miR-181c , and let-7 in CAsE-PE cells correlated with increased target DB01367 oncogenes , RAN , P51159 , Q9UL26 mRNAs , and P01116 protein . Reduced miR-143 , miR-34c-5p , and miR-205 in As-CSC correlated with increased target RAN mRNA , and P01116 , P01111 , and P10301 proteins . The DB01367 / P29323 and PI3K/ P60484 /AKT pathways control cell survival , differentiation , and proliferation , and when dysregulated promote a cancer phenotype . iAs transformation increased expression of activated P29323 kinase in both transformants and altered components of the PI3K/ P60484 /AKT pathway including decreased P60484 and increases in P10415 , BCL-XL , and P15692 in the absence of AKT activation . Thus , dysregulated miRNA expression may be linked to DB01367 activation in both transformants . Stress response genes are suppressed in mouse preimplantation embryos by granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) . BACKGROUND : P04141 ( GM- P04141 ) is known to promote the development and survival of human and mouse preimplantation embryos ; however , the mechanism of action of GM- P04141 in embryos is not defined . METHODS : Mouse blastocysts were cultured from zygote stage in vitro with and without recombinant mouse GM- P04141 ( rmGM- P04141 ) , and in vivo developed blastocysts were flushed from Csf2 null mutant and wild-type mice . The effect of GM- P04141 on blastocyst expression of stress response and apoptosis genes was evaluated by microarray , qPCR and immunochemistry . RESULTS : Microarray analysis of the gene transcription profile showed suppression of stress response and apoptosis gene pathways in blastocysts exposed to rmGM- P04141 in vitro . qPCR analysis confirmed that rmGM- P04141 inhibited expression of heat shock protein ( HSP ) and apoptosis pathway genes Cbl , Hspa5 , Hsp90aa1 , Hsp90ab1 and Gas5 in in vitro blastocysts . Immunocytochemical analysis of HSP 1 ( P0DMV8 /1B ; HSP70 ) , Q07812 , P10415 and TRP53 ( p53 ) in in vitro blastocysts showed that P0DMV8 /1B and P10415 proteins were less abundant when embryos were cultured with rmGM- P04141 . Q07812 and TRP53 were unchanged at the protein level , but Bax mRNA expression was reduced after GM- P04141 treatment . In in vivo developed blastocysts , Csf2 null mutation caused elevated expression of Hsph1 but not other stress response genes . CONCLUSIONS : We conclude that GM- P04141 inhibits the cellular stress response and apoptosis pathways to facilitate embryo growth and survival , and the protective effects of GM- P04141 are particularly evident in in vitro culture media , whereas in vivo other cytokines can partly compensate for absence of GM- P04141 . P10997 -leptin coadministration stimulates central histaminergic signaling in rats . Combined amylin+leptin ( Q9BXJ7 + P41159 ) can reduce diet induced obesity and is very effective in combating P41159 resistance . The purpose of this study was to evaluate the effect of Q9BXJ7 + P41159 on central histaminergic signaling in lean and obese rats . Male rats were administered P41159 ( 300 μg/kg/d ) , Q9BXJ7 ( 100 μg/kg/d ) , Q9BXJ7 + P41159 or vehicle ( SAL , 0.9 % normal saline ) , via a subcutaneous mini-osmotic pump or single injection ( P41159 , 300 μg/kg and Q9BXJ7 , 100 μg/kg ) for acute studies . Q9BXJ7 + P41159 administration increased expression of histamine H1 receptor ( HIR ) and histidine decarboxylase ( HDC ) mRNA in the hypothalamus . Increased levels of P35367 were seen in arcuate ( Arc ) and ventromedial hypothalamus ( VMH ) as well as the area postrema ( APOS ) and nucleus of solitary tract ( P30990 ) following Q9BXJ7 + P41159 administration . APOS and P30990 also showed expression of immediate early gene c- P01100 in the hindbrain in Q9BXJ7 + P41159 -treated rats . We confirmed previous evidence indicating that Q9BXJ7 + P41159 increased P35610 -3 protein phosphorylation in Arc and VMH . Finally , by in vivo microdialysis , we observed an increase in methyl P42357 levels in the VMH of Q9BXJ7 , P41159 and Q9BXJ7 + P41159 -treated rats . Taken together , these observations are consistent with an important role that neuronal P42357 may play in mediating the potent effects of Q9BXJ7 + P41159 on food intake and body weight . Chemokines regulate hippocampal neuronal signaling and gp120 neurotoxicity . The HIV-1 envelope protein gp120 induces apoptosis in hippocampal neurons . Because chemokine receptors act as cellular receptors for HIV-1 , we examined rat hippocampal neurons for the presence of functional chemokine receptors . Fura-2-based Ca imaging showed that numerous chemokines , including SDF-1alpha , RANTES , and fractalkine , affect neuronal Ca signaling , suggesting that hippocampal neurons possess a wide variety of chemokine receptors . Chemokines also blocked the frequency of spontaneous glutamatergic excitatory postsynaptic currents recorded from these neurons and reduced voltage-dependent Ca currents in the same neurons . Reverse transcription-PCR demonstrated the expression of P32246 , CCR4 , P51681 , P51686 /10 , P25025 , P61073 , and P49238 , as well as the chemokine fractalkine in these neurons . Both fractalkine and macrophage-derived chemokine ( MDC ) produced a time-dependent activation of extracellular response kinases ( P29323 ) -1/2 , whereas no activation of c- P05412 NH2-terminal protein kinase ( JNK ) /stress-activated protein kinase , or p38 was evident . Furthermore , these two chemokines , as well as SDF-1alpha , activated the Ca- and DB02527 -dependent transcription factor CREB . Several chemokines were able also to block gp120-induced apoptosis of hippocampal neurons , both in the presence and absence of the glial feeder layer . These data suggest that chemokine receptors may directly mediate gp120 neurotoxicity . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . Chemopreventive effects of the dietary histone deacetylase inhibitor tributyrin alone or in combination with vitamin A during the promotion phase of rat hepatocarcinogenesis . The chemopreventive effects of tributyrin ( TB ) and vitamin A ( VA ) , alone or in combination , were investigated during the promotion phase of rat hepatocarcinogenesis . Compared to diethylnitrosamine control rats , TB and TB+VA-treated rats , but not VA-treated rats , presented a lower incidence and mean number of hepatocyte nodules and a smaller size of persistent preneoplastic lesions ( pPNLs ) . In addition , TB and TB+VA-treated rats exhibited a higher apoptotic body index in pPNL and remodeling PNL , whereas VA-treated rats presented only a higher apoptotic body index in remodeling PNL . None of the treatments inhibited cell proliferation in PNL . TB and TB+VA-treated rats , but not VA-treated rats , exhibited higher levels of H3K9 acetylation and P38936 protein expression . TB and VA-treated rats exhibited increased hepatic concentrations of butyric acid and retinoids , respectively . Compared to normal rats , diethylnitrosamine control animals exhibited lower retinyl palmitate hepatic concentrations . All groups had similar expression levels and exhibited similar unmethylated P09455 promoter region in microdissected pPNL , indicating that epigenetic silencing of this gene was not involved in alteration of retinol metabolism in early hepatocarcinogenesis . Data support the effectiveness of TB as a dietary histone deacetylase inhibitor during the promotion phase of hepatocarcinogenesis , which should be considered for chemoprevention combination strategies . Structure of the human Q9H244 receptor in complex with an antithrombotic drug . P2Y receptors ( P2YRs ) , a family of purinergic G-protein-coupled receptors ( GPCRs ) , are activated by extracellular nucleotides . There are a total of eight distinct functional P2YRs expressed in human , which are subdivided into P47900 -like receptors and Q9H244 -like receptors . Their ligands are generally charged molecules with relatively low bioavailability and stability in vivo , which limits our understanding of this receptor family . P2Y12R regulates platelet activation and thrombus formation , and several antithrombotic drugs targeting P2Y12R -- including the prodrugs clopidogrel ( Plavix ) and prasugrel ( DB06209 ) that are metabolized and bind covalently , and the nucleoside analogue ticagrelor ( DB08816 ) that acts directly on the receptor -- have been approved for the prevention of stroke and myocardial infarction . However , limitations of these drugs ( for example , a very long half-life of clopidogrel action and a characteristic adverse effect profile of ticagrelor ) suggest that there is an unfulfilled medical need for developing a new generation of P2Y12R inhibitors . Here we report the 2.6 Å resolution crystal structure of human P2Y12R in complex with a non-nucleotide reversible antagonist , AZD1283 . The structure reveals a distinct straight conformation of helix V , which sets P2Y12R apart from all other known class A GPCR structures . With AZD1283 bound , the highly conserved disulphide bridge in GPCRs between helix III and extracellular loop 2 is not observed and appears to be dynamic . Along with the details of the AZD1283-binding site , analysis of the extracellular interface reveals an adjacent ligand-binding region and suggests that both pockets could be required for dinucleotide binding . The structure provides essential insights for the development of improved P2Y12R ligands and allosteric modulators as drug candidates . Gq-mediated Akt translocation to the membrane : a novel PIP3-independent mechanism in platelets . Akt is an important signaling molecule regulating platelet aggregation . Akt is phosphorylated after translocation to the membrane through Gi signaling pathways by a phosphatidylinositol-3,4,5-trisphosphate ( PIP3 ) -dependent mechanism . However , Akt is more robustly phosphorylated by thrombin compared with adenosine 5'-diphosphate in platelets . This study investigated the mechanisms of Akt translocation as a possible explanation for this difference . Stimulation of washed human platelets with protease-activated receptor agonists caused translocation of Akt to the membrane rapidly , whereas phosphorylation occurred later . The translocation of Akt was abolished in the presence of a Gq-selective inhibitor or in Gq-deficient murine platelets , indicating that Akt translocation is regulated downstream of Gq pathways . Interestingly , phosphatidylinositol 3-kinase ( PI3K ) inhibitors or Q9H244 antagonist abolished Akt phosphorylation without affecting Akt translocation to the membrane , suggesting that Akt translocation occurs through a PI3K/PIP3/Gi-independent mechanism . An Akt scaffolding protein , P38936 -activated kinase ( PAK ) , translocates to the membrane after stimulation with protease-activated receptor agonists in a Gq-dependent manner , with the kinetics of translocation similar to that of Akt . Coimmunoprecipitation studies showed constitutive association of PAK and Akt , suggesting a possible role of PAK in Akt translocation . These results show , for the first time , an important role of the Gq pathway in mediating Akt translocation to the membrane in a novel Gi/PI3K/PIP3-independent mechanism . Transformed 3T3 cells have reduced levels and altered subcellular distribution of the major PKC substrate protein P29966 . The P29966 ( myristylated alanine-rich C-kinase substrate ) protein is an abundant calmodulin-binding protein that is a major and specific endogenous substrate of protein kinase C ( PKC ) . Stimulation of cells with phorbol esters or other activators of PKC has been shown previously to result in rapid phosphorylation of P29966 proteins and redistribution of these myristylated C-kinase substrates from membrane to cytosol . Here we show that NIH3T3 murine fibroblasts transformed by P38936 -HA-C- DB01367 or pp60-V- P12931 oncoproteins have markedly reduced levels of p68- P29966 and that most of the remaining P29966 protein is found in the cytosol . 3T3 cells containing a nontransforming oncoprotein Q9Y3Q3 - P10415 , in contrast , exhibited normal levels and distribution of p68- P29966 . When taken together with recent evidence that P29966 proteins are involved in regulating organization of the membrane cytoskeleton , our findings suggest that oncoprotein-mediated alterations in P29966 protein levels and subcellular distribution may contribute to the development or maintenance of the transformed phenotype . Genetic analysis of expression profile involved in retinoid metabolism in non-alcoholic fatty liver disease . AIM : The patients with non-alcoholic fatty liver disease ( NAFLD ) have been reported to be at greater risk for progression to chronic liver disease including liver cirrhosis ( LC ) . To examine the mechanisms for the progression of NAFLD , a genetic analysis of hepatic expression profile in retinoid metabolism in NAFLD was performed since the loss of retinoid signaling is associated with the progression of liver disease via reactive oxygen species ( ROS ) generation . METHODS : Fifty-one genes , which are associated with retinoid metabolism and action , were examined in thirty six subjects including 17 patients with simple steatosis , 11 with non-alcoholic steatohepatitis ( NASH ) and eight controls were examined by real-time reverse transcriptase polymerase chain reaction . Immunohistochemical study was also done by 3 kinds of antibodies . RESULTS : Higher expression of P09455 O95237 , DGT1/2 and CES1 in NAFLD suggests that mutual conversion between retinyl ester and retinal occurs actively . Expression of P07327 /2/3 , Q92781 /10/11 , O75911 and RALDH1/3 was increased in NAFLD , suggesting that oxidation process from retinol to all-trans retinoic acid ( DB00755 ) was enhanced . Importantly , greater expression of O43174 indicated that degradation of DB00755 was enhanced in NAFLD . Further , expression of P00441 /2 , catalase , thioredoxin and uncoupling protein 2 was also enhanced . CONCLUSION : Hyperdynamic state of retinoid metabolism is present in the liver tissues with NAFLD , which may be a putative mechanism by which NAFLD progresses to chronic liver disease including LC . Echicetin coated polystyrene beads : a novel tool to investigate GPIb-specific platelet activation and aggregation. P04275 /ristocetin ( P04275 /R ) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin , which also binds P04275 . These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets . Here , we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads , which specifically activate GPIb . We compared platelet activation induced by echicetin beads to P04275 /R . Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling . Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets , while under the same conditions P04275 /R treatment led only to αIIbβ3-independent platelet agglutination . The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation , while the total amount of echicetin used for activation is not critical . Echicetin beads induced strong phosphorylation of several proteins including p38 , P29323 and P31749 . Synergistic signaling via Q9H244 and thromboxane receptor through secreted ADP and TxA2 , respectively , were important for echicetin bead triggered platelet activation . Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation . Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways . The BH3 mimetic DB05764 synergizes with the Q02750 /2 inhibitor selumetinib/AZD6244 to promote O43521 -dependent tumour cell death and inhibit acquired resistance . Tumour cells typically exhibit a G(1) cell cycle arrest in response to the Q02750 /2 [ mitogen-activated protein kinase/ P29323 ( extracellular-signal-regulated kinase ) kinase 1/2 ] inhibitor selumetinib , but do not die , and thus they acquire resistance . In the present study we examined the effect of combining selumetinib with the BH3 [ P10415 ( B-cell lymphoma 2 ) homology domain 3 ] -mimetic P10415 inhibitor DB05764 . Although either drug alone caused little tumour cell death , the two agents combined to cause substantial caspase-dependent cell death and inhibit long-term clonogenic survival of colorectal cancer and melanoma cell lines with P15056 (V600E) or DB01367 mutations . This cell death absolutely required Q07812 ( P10415 -associated X protein ) and was inhibited by RNAi ( RNA interference ) -mediated knockdown of O43521 ( P10415 -interacting mediator of cell death ) in the P15056 (V600E)-positive COLO205 cell line . When colorectal cancer cell lines were treated with selumetinib plus DB05764 we observed a striking reduction in the incidence of cells emerging with acquired resistance to selumetinib . Similar results were observed when we combined DB05764 with the P15056 (V600E)-selective inhibitor PLX4720 , but only in cells expressing P15056 (V600E) . Finally , cancer cells in which acquired resistance to selumetinib arises through P15056 (V600E) amplification remained sensitive to DB05764 , whereas selumetinib-resistant HCT116 cells ( P01116 (G13D) amplification ) were cross-resistant to DB05764 . Thus the combination of a P10415 inhibitor and an P27361 /2 pathway inhibitor is synthetic lethal in P27361 /2-addicted tumour cells , delays the onset of acquired resistance and in some cases overcomes acquired resistance to selumetinib .

MH_train_59

MH_train_59

MH_train_59

interacts_with DB01229?

multiple_choice

Patient age and biological aggressiveness of endometrial carcinoma . BACKGROUND : Advanced age is associated with a significantly worse prognosis of endometrial carcinoma patients . The aim of this study was to test whether age is a poor-risk factor in endometrial carcinoma because tumors arising in older patients are biologically different from those diagnosed in patients of an earlier age . MATERIALS AND METHODS : DB03843 -fixed , paraffin-embedded samples from 136 previously untreated patients with endometrial carcinoma were studied by means of immunohistochemistry . The expression of molecular markers associated with hormone responsiveness ( estrogen and progesterone receptors ) , proliferation ( Ki67 , C-ERB-B2 , p53 ) , invasiveness ( P12830 ) and apoptosis ( P10415 and p53 ) was analyzed . The obtained expression levels , together with all available clinical and pathological features were tested for correlations with the patients age and survival . RESULTS : Advanced patient age showed a direct correlation with tumor stage ( r=0.29 , p=0.0008 ) and mutant p53 expression ( r=0.25 , p=0.004 ) , and an inverse correlation with P12830 expression ( r=-0.28 , p=0.001 ) . Patient age above the 25th percentile ( 57 years ) of the age distribution was significantly associated with a worse prognosis ( p=0.018 ) . CONCLUSION : It appears that with advancing age , endometrial carcinoma exhibits a more aggressive tumor phenotype , characterized by mutant p53 expression and down-regulation of P12830 expression , and that this , in its turn , results in tumors being diagnosed at a more advanced stage in older patients . Production of paired helical filament , tau-like proteins by PC12 cells : a model of neurofibrillary degeneration . Neuron-like cells derived from a rat pheochromocytoma cell line ( PC12 ) and differentiated with nerve growth factor produce a paired helical filament ( PHF ) -like antigen when they are subjected to heat shock ( Wallace et al. : Mol Brain Res 19:149-155 , 1993 ) . It accumulates in a localized region of the perinuclear cytoplasm and reacts with monoclonal antitau antibodies , which identify epitopes in the N- and C-terminal halves and the microtubule-binding domain of tau protein . The observed profile of immunoreactivity suggests the presence of full-length and C-terminally truncated tau in a region of perinuclear cytoplasm in which no structurally intact PHFs could be demonstrated by conventional transmission electron microscopy . The accumulated tau protein colocalized with antibodies raised against mitochondrial outer membrane proteins and was associated with the presence of numerous mitochondrial profiles that were demonstrated with electron microscopy . Because differentiated PC12 cells pretreated with colcemid or DB01229 prior to heat shock fail to exhibit perinuclear PHF-like immunoreactivity , the reported response to heat shock appears to require an intact system of intracellular microtubules . This PC12 system provides a model in which the metabolic and molecular biological underpinnings of neuronal degeneration in Alzheimer 's disease can be manipulated . The system may eventually be applicable to the development of pharmaceutical agents that interfere with formation and/or degeneration of P10636 in Alzheimer 's disease . Monocyte stimulation by reactive oxygen species : role of superoxide and intracellular Ca2+ . OBJECTIVE : Production of reactive oxygen species has generally been linked to inflammatory processes . Whether the presence of superoxide and hydrogen peroxide affects mononuclear cell function was investigated with an in vitro model using isolated human mononuclear cells . MATERIALS AND METHODS : Human mononuclear cells were isolated from buffy coat . Toxicity was measured with DB09158 exclusion , secreted P01375 was measured with an ELISA , reactive oxygen species within mononuclear cells were quantified with dichlorodihydrofluorescein diacetate fluorescence , intracellular P01375 was measured with flow cytometry and fluorescence microscopy . RESULTS : The enzymatic production of superoxide caused a dose-dependent increase in P01375 synthesis , whereas hydrogen peroxide was ineffective . Flow cytometric measurements and immunofluorescence microscopy demonstrated that monocytes were the main P01375 producing population . This proinflammatory reaction was further characterized by pharmacologically investigating Ca2+ signalling pathways . Superoxide stimulated P01375 secretion was inhibited by intracellular Ca2+-buffers ( P10636 -AM and BAPT-AM ) or VOC operating antagonists ( diltiazem and verapamil ) and only to a small extent by pharmacological inhibitors of ligand-gated pathways ( TMB-8 and SKF 96368 ) . CONCLUSION : We propose that superoxide activates human mononuclear cells in a Ca2+-dependent manner . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . O75144 costimulation is required for T-cell encephalitogenicity . The interaction of Q9Y6W8 with its ligand on P25054 provides a costimulatory signal to previously activated T-cells . In these studies , we blocked the Q9Y6W8 : O75144 interaction with Q9Y6W8 -Ig during the in vitro activation of MBP-reactive transgenic P01730 (+) T-cells . The presence of Q9Y6W8 -Ig in these cultures inhibited the ability of the transgenic T-cells to transfer EAE , although they entered the brains of the recipient mice . Q9Y6W8 -Ig increased apoptosis in the transgenic T-cells , especially in the memory population . This enhanced apoptosis was accompanied by an increase in the Q07812 /BCL-2 mRNA ratio . Q9Y6W8 -Ig did not prevent P60568 production , demonstrating that P60568 production is O75144 independent . P01579 and P22301 production by the transgenic T-cells , however , was suppressed . Finally , Q9Y6W8 -Ig injection into mice after the first signs of EAE ameliorated clinical disease . Therefore , O75144 provides a signal distinct from P10747 costimulation that is required for the activation and viability of encephalitogenic T-cells . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Cluster analysis of risk factor genetic polymorphisms in Alzheimer 's disease . Multiple genetic variants may contribute to the risk of developing Alzheimer 's disease . We have analyzed polymorphisms in 9 genes to determine whether particular combinations would contribute to this risk . The genes were P02649 , LDLr , P01034 , P07339 , P01375 , P56817 , P10636 , Q8IWL8 , P29474 , and Q12800 . Three risk groups for the disease were identified . Risk group I was younger , was heterozygous for the P01034 ( GA ) , CTSD2936 ( AG ) , P01375 -308 ( AG ) genetic variants . Risk group II was older , was homozygous for the -427 P02649 promoter polymorphism ( TT ) , and heterozygous for the P10636 deletion and for the Q8IWL8 variant ( QR ) . Group III had both the youngest and oldest subjects , were heterozygous for the -863 ( AC ) and -1031 ( CT ) P01375 promoter polymorphisms . All three groups carried the P02649 4 allele and were heterozygous for both P56817 polymorphisms . The control groups were carriers of the P02649 3 allele and were homozygous for the P56817 genetic variants . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Q03135 tyrosine phosphorylation enhances paclitaxel-mediated cytotoxicity . Q03135 ( Q03135 ) , a highly conserved membrane-associated protein , is a putative regulator of cellular transformation . Q03135 is localized in the plasmalemma , secretory vesicles , Golgi , mitochondria , and endoplasmic reticulum membrane and associates with the microtubule cytoskeleton . Taxanes such as paclitaxel ( DB01229 ) are potent anti-tumor agents that repress the dynamic instability of microtubules and arrest cells in the G(2)/M phase . Src phosphorylation of DB00135 -14 on Q03135 regulates its cellular localization and function . We report that phosphorylation of Q03135 on DB00135 -14 regulates paclitaxel-mediated apoptosis in MCF-7 breast cancer cells . Befitting its role as a multitasking molecule , we show that Q03135 sensitizes cells to apoptosis by regulating cell cycle progression and activation of the apoptotic signaling molecules P10415 , p53 , and P38936 . We demonstrate that phosphorylated Q03135 triggers apoptosis by inactivating P10415 and increasing mitochondrial permeability more efficiently than non-phosphorylated Q03135 . Furthermore , expression of P38936 , which correlates with taxane sensitivity , is regulated by Q03135 phosphorylation in a p53-dependent manner . Collectively , our findings underscore the importance of Q03135 phosphorylation in apoptosis and suggest that events that negate Q03135 tyrosine phosphorylation may contribute to anti-microtubule drug resistance . Enhanced sensitization to taxol-induced apoptosis by herceptin pretreatment in ErbB2-overexpressing breast cancer cells . The recombinant humanized anti-ErbB2/ P04626 monoclonal antibody Herceptin ( DB00072 ) has been shown to significantly enhance the tumoricidaleffects of antitumor drugs such as paclitaxel ( DB01229 ) in patients with ErbB2-overexpressing breast cancers . Here , we investigated the molecular mechanisms by which Herceptin enhances the antitumor effects of DB01229 . Because activation of p34(Cdc2) is required for DB01229 -induced apoptosis and because overexpression of ErbB2 blocks DB01229 -induced apoptosis by inhibiting p34(Cdc2) activation , we studied the effect of Herceptin treatment on p34(Cdc2) kinase activation and apoptosis in DB01229 -treated human breast carcinoma cell lines MDA-MB-435 , SKBr3 , MDA-MB-453 , and 435.eB , which is an ErbB2 transfectant of MDA-MB-435 . Herceptin treatment down-regulated ErbB2 , reduced the inhibitory phosphorylation of Cdc2 on DB00135 -15 , and down-regulated the expression of P38936 (Cip1) , a Cdc2 inhibitor . Herceptin plus DB01229 treatment led to higher levels of p34(Cdc2) kinase activity and apoptosis in ErbB2-overexpressing breast cancer cells , which is likely attributable to inhibition of Cdc2- DB00135 -15 phosphorylation and P38936 (Cip1) expression . Because significant dephosphorylation of Cdc2- DB00135 -15 and down-regulation of P38936 (Cip1) occur at least 24 h after Herceptin treatment , we investigated whether 24 h Herceptin pretreatment will render ErbB2-overexpressing breast cancer cells more sensitive to DB01229 -induced apoptosis compared with the simultaneous treatment of Herceptin plus DB01229 . Indeed , Herceptin pretreatment increased DB01229 -induced apoptosis and cytotoxicity in vitro and more effectively inhibited the growth of tumor xenografts with enhanced in vivo apoptosis . Thus , Herceptin treatment of ErbB2-overexpressing cells can inhibit ErbB2-mediated Cdc2- DB00135 -15 phosphorylation and P38936 (Cip1) up-regulation , which allows effective p34(Cdc2) activation and induction of apoptosis upon DB01229 treatment . Herceptin pretreatment renders ErbB2-overexpressing breast cancers more susceptible to DB01229 -induced cell death , which may have important clinical therapeutic implications . Molecular response of HL-60 cells to mitotic inhibitors vincristine and taxol visualized with apoptosis-related gene expressions , including the new member Q9HB09 . DB01229 and vincristine belong to a group of anticancer drugs that target microtubules , subsequently arresting cells at the mitotic phase of the cell cycle and inducing programmed cell death . The P10415 ( bcl-2 ) family of genes is of known implication in apoptosis induced by various stimuli , among which Q9HB09 , a new member of the family , cloned by our group . For further insights into the mechanisms and molecular targets implicated and modified as a result of apoptosis induced by these two mitosis-arresting drugs , we studied the possible alterations , at the mRNA level , of various apoptosis-related genes ( P10415 , Q07812 , Q9HB09 , CASPASE-3 , FAS ) after leukemia cell ( HL-60 ) treatment with these drugs . The kinetics of cell toxicity were evaluated by the MTT [ 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ] method , trypan blue staining , and cell proliferation efficiency ; apoptosis induction was assayed by endonucleosomal cleavage of DNA ( DNA laddering ) ; and the expression levels of the genes were analysed by RT-PCR , using gene-specific primers . The percentage of nonviable cells was upregulated with increasing cell exposure time and drug concentrations to both taxol and vincristine . Distinct modulations of apoptosis-related genes at the mRNA level were also observed , mainly concerning P10415 and Q9HB09 along apoptosis induction . Our results indicate and support the hypothesis that the apoptosis-related genes P10415 and Q9HB09 respond similarly to treatment of the human , acute , myelocytic leukemia HL60 cells with the anticancer drugs vincristine and taxol though in a drug-specific and time-dependent manner . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . Endothelial [Ca2+]i and caveolin-1 antagonistically regulate P29474 activity and microvessel permeability in rat venules . AIMS : In this study , we investigated the mechanisms by which caveolin-1 ( Q03135 ) inhibits increases in permeability induced by platelet activating factor ( Q15004 ) and elucidated the relationship between the endothelial intracellular Ca(2+) concentration ( [Ca(2+)](i) ) and Q03135 in regulating endothelial nitric oxide synthase ( P29474 ) activity and microvessel permeability in intact microvessels . METHODS AND RESULTS : Experiments were conducted in individually perfused mesenteric venules in Sprague-Dawley rats . Permeability was determined by measuring hydraulic conductivity ( Lp ) . Endothelial [Ca(2+)](i) and nitric oxide ( NO ) production were measured in fura-2- and P08174 -2-loaded microvessels . Perfusion of the Q03135 scaffolding domain , AP- Q03135 , at 1 microM for 30 min did not affect Q15004 -induced increases in endothelial [Ca(2+)](i) but significantly attenuated Q15004 -induced NO production from 143 +/- 2 to 110 +/- 3 % of control fluorescence intensity ( FI ) . The Q15004 -induced Lp increase was correlatively reduced from a mean peak value of 7.5 +/- 0.9 to 1.9 +/- 0.5 times that of the control . Increasing extracellular [ Ca(2+) ] that potentiated Q15004 -induced peak [Ca(2+)](i) from 500 to 1225 nM augmented NO production to 193 +/- 13 % and further increased Lp to 17.3 +/- 1.6 times the control value . More importantly , enhanced Ca(2+) influx restored the reduced NO production and Lp by AP- Q03135 with NO FI at 149 % and Lp at 7.7 +/- 1.1 times the control value . CONCLUSION : Our results indicate that P29474 inhibition and reduced NO production contribute to the inhibitory action of AP- Q03135 on Q15004 -induced increases in permeability . Q03135 and endothelial [Ca(2+)](i) antagonistically regulate P29474 activity in intact microvessels , and the level of produced NO is the key determinant of the degree of permeability increases during inflammation . Exposure to an organophosphate ( DB00677 ) during a defined period in neonatal life induces permanent changes in brain muscarinic receptors and behaviour in adult mice . The organophosphate DB00677 ( DB00677 ) is a well-known inhibitor of cholinesterases . We have recently observed that neonatal exposure to a single subsymptomal dose of DB00677 induces permanent alterations in muscarinic cholinergic receptors ( MAChRs ) and in spontaneous behaviour , in the mice as adults . In order to determine if there is a critical period for these effects , neonatal mice were given a single oral dose of 1.5 mg/kg DB00677 b.wt. on postnatal day 3 , 10 or 19 , causing equal inhibition of P22303 . At the adult age of 4 months the mice were tested for spontaneous motor behaviour , and were subsequently sacrificed for measurement of density of MAChRs and subpopulations of MAChRs in the cerebral cortex by using the antagonist quinuclidinyl benzilate ( [3H]QNB ) , and agonist carbachol , respectively . At adult age , mice exposed to DB00677 on postnatal day ( P01160 ) 3 or 10 showed significant ( P < or = 0.01 ) alterations in spontaneous motor behaviour and a significant ( P < or = 0.01 ) decrease in muscarinic receptor density . There were no alterations mice exposed on P01160 19 . The proportions and affinity-constants of high- and low-affinity MAChR binding sites were not affected in mice showing altered MAChR density . The lack of effect on mice exposed on P01160 19 was not due to differences in P22303 activity . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Regulation of the human P38936 (waf1/cip1) gene promoter via multiple binding sites for p53 and the vitamin D3 receptor . The main regulator of the human tumor suppresser gene P38936 (waf1/cip1) is the transcription factor p53 , but more recently it has been suggested to be a primary anti-proliferative target for the nuclear receptor P11473 in the presence of its ligand 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) . To identify P11473 responding regions , we analyzed 20 overlapping regions covering the first 7.1 kb of the P38936 (waf1/cip1) promoter in MCF-7 human breast cancer cells using chromatin immuno-precipitation assays ( ChIP ) with antibodies against p53 and P11473 . We confirmed two known p53 binding regions at approximate positions -1400 and -2300 and identified a novel site at position -4500 . In addition , we found three P11473 -associated promoter regions at positions -2300 , -4500 and -6900 , i.e. two regions showed binding for both p53 and P11473 . In silico screening and in vitro binding assays using recombinant and in vitro translated proteins identified five p53 binding sites within the three p53-positive promoter regions and also five DB00136 response elements within the three P11473 -positive regions . Reporter gene assays confirmed the expected responsiveness of the respective promoter regions to the p53 inducer 5-fluorouracil and DB00136 . Moreover , re-ChIP assays confirmed the functionality of the three DB00136 -reponsive promoter regions by monitoring simultaneous occupancy of P11473 with the co-activator proteins CBP , Q15788 and Q15648 . Taken together , we demonstrated that the human P38936 ( ( waf1/cip1 ) ) gene is a primary DB00136 -responding gene with at least three P11473 binding promoter regions , in two of which also p53 co-localizes . DB00820 , a further innovation in the treatment of sexual dysfunction . In recognition of the large number of sufferers of sexual dysfunction worldwide , and the variety of etiologies of the condition , investigation into effective pharmacological agents has been expanded . One method of intervention is inhibition of the phosphodiesterase type 5 ( O76074 ) enzyme , which has already been exploited with a considerable degree -- though not complete -- success . A number of new agents that inhibit O76074 are under development . Notable among these is tadalafil , which has demonstrated a high level of selectivity for O76074 over the other phosphodiesterases and has shown efficacy in improving erectile function and sexual satisfaction in phase III trials . Throughout the clinical development program for tadalafil , the drug has been well tolerated and without serious side effects . The manufacturer , Lilly Q9Y6W8 , received an approvable letter from the US Food and Drug Administration for use of the drug as a treatment for erectile dysfunction on April 30 , 2002 . Lilly Q9Y6W8 hopes to market tadalafil , with the trade name DB00820 , in the USA in 2003 . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Gene network profiling before and after transplantation in alcoholic cirrhosis liver transplant recipients . The main objective of this study was to define a gene network profile network in liver transplant recipients with alcoholic cirrhosis before and after liver transplantation . Genes were selected from data obtained in a previous study of liver transplant recipients with alcoholic cirrhosis . Selected up-regulated genes were further validated by quantitative real-time polymerase chain reaction in different groups of liver transplant recipients with alcoholic cirrhosis ( n=5 ) . Selected genes up-regulated before transplantation were : Q07011 ( tumor necrosis factor [ P01375 ] receptor superfamily , member 9 ) ; P14784 ( interleukin-2 receptor beta ) ; Q92843 ( P10415 -like 2 ) ; Q96PH1 ( NADPH ) oxidase , EF-hand calcium binding domain 5 ) ; P50542 ( peroxisomal biogenesis factor 5 ) ; P37231 ( peroxisome proliferator-activated receptor gamma ) ; Q96Q05 ( O14920 binding protein ) ; Q9NYR9 ( NFKappaBeta inhibitor interacting Ras-like 2 ) ; P05112 ( interleukin-4 ) ; IL-4R ( interleukin 4 receptor ) ; P07327 ( alcohol dehydrogenase 1A , class 1 ) ; O75891 ( aldehyde dehydrogenase 1 family , member Q9NUQ9 ) ; P05164 ( myeloperoxidase ) ; P01160 ( natriuretic peptide precursor A ) ; Q16548 ( P10415 -related protein A1 ) ; P24522 ( growth arrest and DNA-damage-inducible alpha ) ; P55061 ( P55061 ) ; P42336 ( phosphoinositide-3-kinase , catalytic , alpha polypeptide ) ; P38484 ( interferon gamma receptor 2 ) ; O60674 ( Janus Kinase 2 ) ; FAS ( Fas , P01375 receptor superfamily , member 6 ) ; Q92844 ( TRAF family member-associated NFKB activator ) ; O95551 ( O95551 ) ; and P08758 ( annexin A5 ) . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone .

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MH_train_60

interacts_with DB00005?

multiple_choice

Pharmacokinetics and brain uptake of an IgG- P01375 decoy receptor fusion protein following intravenous , intraperitoneal , and subcutaneous administration in mice . P01375 ( P01375 ) -α is a proinflammatory cytokine active in the brain . DB00005 , the P01375 decoy receptor ( TNFR ) , does not cross the blood-brain barrier ( BBB ) . The TNFR was re-engineered for BBB penetration as a fusion protein with a chimeric monoclonal antibody ( mAb ) against the mouse transferrin receptor ( P02786 ) , and this fusion protein is designated cTfRMAb-TNFR . The cTfRMAb domain of the fusion protein acts as a molecular Trojan horse and mediates transport via the endogenous BBB P02786 . To support future chronic treatment of mouse models of neural disease with daily administration of the cTfRMAb-TNFR fusion protein , a series of pharmacokinetics and brain uptake studies in the mouse was performed . The cTfRMAb-TNFR fusion protein was radiolabeled and injected into mice via the intravenous , intraperitoneal ( IP ) , or subcutaneous ( SQ ) routes of administration at doses ranging from 0.35 to 10 mg/kg . The distribution of the fusion protein into plasma following the IP or SQ routes was enhanced by increasing the injection dose from 3 to 10 mg/kg . The fusion protein demonstrated long circulation times with high metabolic stability following the IP or SQ routes of injection . The IP or SQ routes produced concentrations of the cTfRMAb-TNFR fusion protein in the brain that exceed by 20- to 50-fold the concentration of TNFα in pathologic conditions of the brain . The SQ injection is the preferred route of administration , as the level of cTfRMAb fusion protein produced in the brain is comparable to that generated with intravenous injection , and at a much lower plasma area under the concentration curve of the fusion protein as compared to IP administration . Reduction of urinary levels of pyridinoline and deoxypyridinoline and serum levels of soluble receptor activator of NF-kappaB ligand by etanercept in patients with rheumatoid arthritis . The effects of soluble P01375 receptor , etanercept , on bone metabolism were investigated in patients with rheumatoid arthritis ( RA ) . Thirty RA patients were administered etanercept once or twice a week for more than 6 months . We evaluated clinical and laboratory parameters and measured urinary excretion levels of pyridinoline ( PYD ) , deoxypyridinoline ( Q12882 ) , cross-linked N-telopeptides of type I collagen ( NTX ) , and serum levels of bone alkaline phosphatase ( BAP ) , osteoprotegerin ( O00300 ) , and soluble receptor activator of NFkappaB ligand ( sRANKL ) at the baseline and at 3 and 6 months after initial treatment with etanercept . DB00005 treatment resulted in an improvement of symptoms due to RA and in a reduction of urinary excretion levels of PYD and Q12882 as well as serum sRANKL levels , with a significant difference at 6 months , and an increase of serum BAP levels at 3 and 6 months after the initial treatment with etanercept . Urinary NTX and serum O00300 levels did not show a significant change at 3 and 6 months after the initial treatment , but serum O00300 levels did show a reverse correlation with serum CRP levels , suggesting that the regulation of inflammation in RA may result in an induction of O00300 production . DB00005 may have the ability to reduce the levels of bone resorption markers and to increase the levels of a bone formation marker while reducing sRANKL formation in RA patients . DB00005 therapy-associated acute uveitis : a case report and literature review . A female patient diagnosed with ankylosing spondylitis experienced a new onset acute iritis following the initiation of etanercept therapy and recurrent episodes of iritis continues during the treatment of etanercept . DB00005 -associated iritis was suspected . Anti- P01375 therapies can alleviate uveitis in some studies , but in some other anecdotal reports etanercept is considered as the main cause of uveitis . A literature review is presented below . For clinicians , more attention must be paid to the potential association between uveitis or iritis and etanercept , and more careful surveillance of patients under etanercept treatment is necessary . P01375 signaling : early events and phosphorylation . P01375 -alpha ( P01375 ) is a major mediator of apoptosis as well as immunity and inflammation . Inappropriate production of P01375 or sustained activation of P01375 signaling has been implicated in the pathogenesis of a wide spectrum of human diseases , including cancer , osteoporosis , sepsis , diabetes , and autoimmune diseases such as multiple sclerosis , rheumatoid arthritis , and inflammatory bowel disease . P01375 binds to two specific receptors , P01375 -receptor type I ( P19438 , CD120a , p55/60 ) and P01375 -receptor type II ( P01375 -R2 , DB00005 , p75/80 ) . Signaling through P19438 is extremely complex , leading to both cell death and survival signals . Many findings suggest an important role of phosphorylation of the P19438 by number of protein kinases . Role of P01375 -R2 phosphorylation on its signaling properties is understood less than P19438 . Other cellular substrates as Q15628 adaptor protein , TRAF protein family and RIP kinases are reviewed in relation to P01375 receptor-mediated apoptosis or survival pathways and regulation of their actions by phosphorylation . P01375 receptor and Fas signaling mechanisms . Four members of the tumor necrosis factor ( P01375 ) ligand family , P01375 , P01374 , Q06643 , and O43557 , interact with four receptors of the P01375 /nerve growth factor family , the p55 P01375 receptor ( CD120a ) , the p75 P01375 receptor ( DB00005 ) , the DB05833 ( LT beta R ) , and herpes virus entry mediator ( Q92956 ) to control a wide range of innate and adaptive immune response functions . Of these , the most thoroughly studied are cell death induction and regulation of the inflammatory process . Fas/Apo1 ( CD95 ) , a receptor of the P01375 receptor family activated by a distinct ligand , induces death in cells through mechanisms shared with CD120a . The last four years have seen a proliferation in knowledge of the proteins participating in the signaling by the P01375 system and CD95 . The downstream signaling molecules identified so far -- caspases , phospholipases , the three known mitogen activated protein ( Q96HU1 ) kinase pathways , and the NF-kappa B activation cascade -- mediate the effects of other inducers as well . However , the molecules that initiate these signaling events , including the death domain- and P01375 receptor associated factor ( TRAF ) domain-containing adapter proteins and the signaling enzymes associated with them , are largely unique to the P01375 /nerve growth factor receptor family . Refractory erythrodermic psoriasis in a child with an excellent outcome by using etanercept . Psoriasis affects 0.12 % to 0.71 % of all children . Erythrodermic psoriasis is an uncommon but serious disorder , occurring in less than 1.5 % of cases . P01375 -alpha blockers ( P01375 -α ) are a new class of drugs used to treat moderate to severe psoriasis refractory to conventional therapies . DB00005 is a TNFα receptor fusion protein , approved by the FDA for treating juvenile rheumatoid arthritis . We present the case of a 7-year-old suffering from plaque psoriasis since 8 months old which evolved into erythroderma refractory to cyclosporine and methotrexate . Patient responded excellently to etanercept , with no adverse side effects . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Therapeutic hotline . Treatment of pityriasis rubra pilaris with etanercept . Pityriasis rubra pilaris ( PRP ) or Devergie 's disease is a chronic and rare papulosquamous disorder of unknown etiology characterized by reddish orange scaly plaques , palmoplantar keratoderma , and keratotic follicular papules . The present authors report a case of a 30-year-old woman with clinical and histologic signs of PRP ( type I adult onset , Griffith 's classification ) . After a few unsuccessful treatments , the present authors chose to start etanercept . Total clearing of the lesions was achieved 5 months after starting the drug . DB00005 is a P01375 -α inhibitor , and today it is largely used in the treatment of several dermatological diseases through blockage of the inflammatory cytokine . The true mechanism of action in PRP remains to be explained ; however , the favorable results in our case raise new questions about P01375 -α 's role in PRP and suggest a therapeutic alternative for resistant cases to classic treatments . To date , there are only three case reports of PRP treated with etanercept in the literature . Vasculoprotective effects of anti-tumor necrosis factor-alpha treatment in aging . Vascular aging is associated with dysregulation of tumor necrosis factor ( P01375 ) -alpha expression . P01375 is a master regulator of vascular proatherogenic phenotypic changes , and it has been linked to endothelial dysfunction and apoptosis . To test the hypothesis that anti- P01375 treatment exerts vasculoprotective effects in aging , aged ( 29 months old ) F344 rats were treated with etanercept ( 1 mg/kg/week for 4 weeks ) , which binds and inactivates P01375 . In aged carotid arteries , relaxations to acetylcholine were decreased , and endothelial O2* production was increased ( as shown by dihydroethidine fluorescence measurements ) . DB00005 treatment significantly improved responses to acetylcholine and decreased vascular NAD(P)H oxidase activity and expression . In aged carotid and coronary arteries , there were increases in DNA fragmentation rate and caspase 3/7 activity ( indicating an increased rate of apoptotic cell death ) , which were attenuated by etanercept treatment . In aged vessels , there was an up-regulation of inflammatory markers , including inducible nitric-oxide synthase and intercellular adhesion molecule-1 , which was decreased by etanercept treatment . In carotid arteries of young animals , recombinant P01375 elicited endothelial dysfunction , oxidative stress , and increased apoptosis and proinflammatory gene expression , mimicking many of the symptoms of vascular aging . Thus , we propose that anti- P01375 treatment exerts anti-aging vasculoprotective effects . DB00005 provides an effective , safe and flexible short- and long-term treatment regimen for moderate-to-severe psoriasis : a systematic review of current evidence . The treatment of psoriasis requires long-lasting intervention . Conventional treatments for psoriasis comprise topical , phototherapeutic and systemic modalities , such as methotrexate or cyclosporine . Biological therapies are advocated by treatment guidelines for the use in moderate-to-severe psoriasis , when conventional treatments have failed , are contraindicated or are associated with severe adverse events . DB00005 is an anti- P01375 recombinant fusion protein that has emerged as a standard biologic treatment option for moderate-to-severe psoriasis . The present review summarizes data from pivotal and post-marketing randomized controlled etanercept trials to treat moderate-to-severe psoriasis for 24 weeks and longer . During the first 12 weeks , etanercept can be administered in different dosing regimens : 50 mg twice weekly ( BIW ) and 50 mg once weekly . Although both regimens are effective , it has been shown that the 50 mg BIW dosage leads to higher response rates at week 24 . In addition , after 24 weeks ' treatment etanercept provides the unique possibility of continuous or intermittent long-term treatment programmes . The medium- to long-term efficacy of etanercept was consistent , regardless of whether etanercept therapy was interrupted or continuous . Taking the chronic nature of psoriasis into account , this flexibility in dosing regimen bestows a key advantage in facilitating individualisation of long-term treatment according to patient needs . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . DB00005 attenuates TNBS-induced experimental colitis : role of P01375 -α expression . Crohn 's disease ( CD ) is associated with gut barrier dysfunction . Tumour necrosis factor-α ( P01375 -α ) plays an important role into the pathogenesis of several inflammatory diseases because its expression is increased in inflamed mucosa of CD patients . Anti- P01375 therapy improves significantly mucosal inflammation . Thus , this study aimed to evaluate the effect of DB00005 ( ETC ) , a tumour necrosis factor alpha ( P01375 -α ) antagonist on the 2,4,6-trinitrobenzene sulfonic acid ( TNBS ) -induced experimental colitis . A total of 18 Wistar rats were randomized into four groups , as follows : ( 1 ) Sham : sham induced-colitis ; ( 2 ) TNBS : non-treated induced-colitis ; ( 3 ) ETC control ; ( 4 ) ETC-treated induced-colitis . Rats from group 4 presented significant improvement either of macroscopic or of histopathological damage in the distal colon . The gene expression of P01375 -α mRNA , decreased significantly in this group compared to the TNBS non-treated group . The treatment with etanercept attenuated the colonic damages and reduced the inflammation caused by TNBS . Taken together , our results suggest that ETC attenuates intestinal colitis induced by TNBS in Wistar rats by P01375 -α downregulation . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Cardiovascular effects of tumour necrosis factor α antagonism in patients with acute myocardial infarction : a first in human study . OBJECTIVE : The inflammatory cytokine , tumour necrosis factor α ( P01375 -α ) , exerts deleterious cardiovascular effects . We wished to determine the effects of P01375 -α antagonism on endothelial function and platelet activation in patients with acute myocardial infarction . DESIGN AND SETTING AND PATIENTS : A double-blind , parallel group , randomised controlled trial performed in a tertiary referral cardiac centre . 26 patients presenting with acute myocardial infarction randomised to receive an intravenous infusion of etanercept ( 10 mg ) or saline placebo . MAIN OUTCOME MEASURES : Leucocyte count , plasma cytokine concentrations , flow cytometric measures of platelet activation and peripheral vasomotor and fibrinolytic function were determined before and 24 h after study intervention . RESULTS : Consistent with effective conjugation of circulating P01375 -α , plasma P01375 -α concentrations increased in all patients following etanercept ( 254 ± 15 vs 0.12 ± 0.02 pg/ml ; p < 0.0001 ) , but not saline infusion . DB00005 treatment reduced neutrophil ( 7.4 ± 0.6 vs 8.8 ± 0.6 × 10(9) cells/l ; p = 0.03 ) and plasma interleukin-6 concentrations ( 5.8 ± 2.0 vs 10.6 ± 4.0 pg/ml ; p = 0.012 ) at 24 h but increased platelet-monocyte aggregation ( 30 ± 5 vs 20 ± 3 % ; p = 0.02 ) . Vasodilatation in response to DB05875 , acetylcholine and sodium nitroprusside , and acute tissue plasminogen activator release were unaffected by either treatment ( p > 0.1 for all ) . CONCLUSIONS : Following acute myocardial infarction , etanercept reduces systemic inflammation but increases platelet activation without affecting peripheral vasomotor or fibrinolytic function . We conclude that P01375 -α antagonism is unlikely to be a beneficial therapeutic strategy in patients with acute myocardial infarction . Tumefactive demyelinating lesions during etanercept treatment requiring decompressive hemicraniectomy . P01375 alpha ( P01375 -α ) is a pro-inflammatory and immunoregulatory cytokine involved in the pathogenesis of several autoimmune disorders . DB00005 , a P01375 -α antagonist ( anti- P01375 -α ) acting as a soluble P01375 -α receptor , has been associated with neurological demyelinating disorders . This paper aims to report an unusual case showing tumefactive central nervous system ( CNS ) inflammatory demyelination in a patient in the course of P01375 -α antagonist therapy , requiring decompressive hemicraniectomy . This report is based on magnetic resonance imaging ( Q9BWK5 ) findings and histology . A biopsy confirmed the inflammatory demyelinating nature of the lesions . The clinical presentation is unusual due to the severity of the disease process , requiring decompressive hemicraniotomy with a clinically favorable outcome . Targeting anti-inflammatory treatment can ameliorate injury-induced neuropathic pain . P01375 -α plays important roles in immune system development , immune response regulation , and T-cell-mediated tissue injury . The present study assessed the net value of anti-tumor necrosis factor-α treatment in terms of functional recovery and inhibition of hypersensitivity after peripheral nerve crush injury . We created a right sciatic nerve crush injury model using a Sugita aneurysm clip . Animals were separated into 3 groups : the first group received only a skin incision ; the second group received nerve crush injury and intraperitoneal vehicle injection ; and the third group received nerve crush injury and intraperitoneal etanercept ( 6 mg/kg ) . DB00005 treatment improved recovery of motor nerve conduction velocity , muscle weight loss , and sciatic functional index . Plantar thermal and von Frey mechanical withdrawal thresholds recovered faster in the etanercept group than in the control group . On day 7 after crush injury , the numbers of ED-1-positive cells in crushed nerves of the control and etanercept groups were increased compared to that in the sham-treated group . After 21 days , ED-1-positive cells had nearly disappeared from the etanercept group . DB00005 reduced expression of interleukin-6 and monocyte chemotactic and activating factor-1 at the crushed sciatic nerve . These findings demonstrate the utility of etanercept , in terms of both enhancing functional recovery and suppressing hypersensitivity after nerve crush . DB00005 does not impede the onset or progression of Wallerian degeneration , but optimizes the involvement of macrophages and the secretion of inflammatory mediators . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . P01375 alpha rapidly activates the mitogen-activated protein kinase ( MAPK ) cascade in a MAPK kinase kinase-dependent , c- P04049 -independent fashion in mouse macrophages . P01375 alpha ( P01375 alpha ) is bound by two cell surface receptors , CD120a ( p55 ) and DB00005 ( p75 ) , that belong to the P01375 /nerve growth factor receptor family and whose signaling is initiated by receptor multimerization in the plane of the plasma membrane . The initial signaling events activated by receptor crosslinking are unknown , although activation of the mitogen-activated protein kinase ( MAPK ) cascade occurs shortly after ligand binding to CD120a . In this study , we investigated the upstream kinases that mediate the activation of the 42-kDa MAPK p42mapk/erk2 following crosslinking of CD120a in mouse macrophages . Exposure of mouse macrophages to P01375 alpha stimulated a time-dependent increase in the activity of MAPK/ P29323 kinase ( MEK ) that temporally preceded peak activation of p42mapk/erk2 . MEKs , dual-specificity threonine/tyrosine kinases , act as a convergence point for several signaling pathways including Ras/Raf , MEK kinase ( Q13233 ) , and Mos . Incubation of macrophages with P01375 alpha was found to transiently stimulate a Q13233 that peaked in activity within 30 sec of exposure and progressively declined toward basal levels by 5 min . By contrast , under these conditions , activation of either c- P04049 or Raf-B was not detected . These data suggest that the activation of the MAPK cascade in response to P01375 alpha is mediated by the sequential activation of a Q13233 and a MEK in a c- P04049 - and Raf-B-independent fashion . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . Clinical experience with soluble P01375 p75 receptor in rheumatoid arthritis . OBJECTIVES : Managing rheumatoid arthritis ( RA ) can be difficult : the disease may follow an unpredictable course , and therapies are often ineffective or toxic . DB00005 , a bioengineered fusion protein derived from the human soluble tumor necrosis factor p75 receptor , recently has been approved for use in patients with refractory RA . METHODS : Published data on clinical experience with etanercept in conjunction with case illustrations are presented . RESULTS : Reports from clinical trials of patients with refractory RA indicate that etanercept significantly improves measures of disease activity , including swollen and tender joint counts , morning stiffness , pain , and erythrocyte sedimentation rate , compared with placebo . In the phase 3 trial , swollen joint counts improved by 47 % in patients receiving etanercept 25 mg compared with a 7 % worsening in patients receiving placebo . The drug is well tolerated ; injection site reaction , the most frequent adverse event , was minor and manageable . In long-term studies , etanercept remains well tolerated and effective . Our clinical experience indicates that patients with refractory RA experience dramatic symptomatic relief , along with reduced fatigue and improved quality of life . One of 18 patients discontinued treatment ; the rest have remained on therapy for up to 18 months . CONCLUSIONS : DB00005 diminishes disease activity in patients with refractory RA . Its favorable safety profile provides symptom control without major toxicity . DB00005 is an important addition to RA therapeutic agents . DB00005 . BACKGROUND : In some patients with psoriasis the inflammatory process fueling skin lesions also afflicts their joints in a condition called psoriatic arthritis . P01375 has been shown to play a central role in the pathogenesis of both cutaneous and joint disease . DB00005 is a soluble fusion protein that binds P01375 , rendering the molecule inactive and making etanercept an effective , targeted therapeutic option for many P01375 -mediated inflammatory diseases . OBJECTIVE : To review the key clinical trials which evaluated efficacy of etanercept for the treatment of psoriasis and psoriatic arthritis , discuss various off-label uses for this therapeutic agent and describe the adverse events associated with its use . METHODS : A search of Medline for relevant articles on etanercept and psoriasis . RESULTS/CONCLUSION : DB00005 has demonstrated efficacy in the treatment of skin and joint manifestations of psoriatic disease , thus gaining FDA approval for use in both . A growing number of off-label dermatological uses for etanercept have been proposed , with variable success reported in individual cases or small series . Since its introduction little over a decade ago , etanercept has maintained a favorable safety profile . Managing moderate-to-severe psoriasis in the elderly . Managing psoriasis in the elderly can be difficult for physicians , who must consider comorbidities , the resulting polypharmacy , and progressive functional impairment of several organs . Indeed , topical agents are the first-line treatment for limited disease . Phototherapy is recommended if topical drugs are not sufficient and the patient has multiple comorbidities and risk factors that make them a poor candidate for an oral or injectable systemic agent . The most important pharmacokinetic alteration in the elderly population is the decreased excretory capacity of the kidney ; thus , cyclosporine should be considered a last resort treatment , and the administered dose of methotrexate should be lowered according to the reduction in estimated creatinine clearance . Acitretin can be used in the absence of severe renal insufficiency , paying attention to lipid profile , treating eventual hyperlipidemia , and closely monitoring liver enzymes . Available data on biological drugs in the elderly are limited . Biologics are associated with a small but significant overall risk of infections . However , there is no convincing evidence that the relative risk of infection with anti-tumor necrosis factor ( P01375 ) -α therapy increases with age . Nevertheless , the package inserts for biologics recommend caution when administering these medications to the geriatric population , due to the high baseline risk of infection in such patients . DB00005 seems to be well tolerated , possibly because of its lower immunosuppressive characteristics compared with other biologics . However , studies with larger sample sizes are needed to confirm its safety . DB00005 decreases synovial expression of tumour necrosis factor-α and lymphotoxin-α in rheumatoid arthritis . OBJECTIVES : DB00005 is an effective tumour necrosis factor ( P01375 ) -α inhibitor drug with the unique ability to block not only P01375 -α but also lymphotoxin ( LT ) -α , at least in vitro . We aimed to investigate the in vivo effect of etanercept on synovial expression of P01375 -α and LT-α . METHOD : Synovial biopsies from 12 rheumatoid arthritis ( RA ) patients started on etanercept and 11 RA patients started on infliximab were obtained at baseline and 8 weeks after treatment initiation . Synovial expression of P01375 -α and LT-α was evaluated by immunohistochemistry followed by computer-assisted image analysis . Differences between paired samples were analysed by the Wilcoxon test and between groups by the Mann-Whitney test . A p-value < 0.05 was considered statistically significant . RESULTS : Six out of the 12 of the patients started on etanercept achieved an American College of Rheumatology ( P10323 )50 response . Macroscopic evaluation of the joints during arthroscopy revealed a significant decrease of local inflammation mainly in good ACR50 responders . Synovial expression of both LT-α and P01375 -α decreased but the differences did not reach statistical significance at a group level . By contrast , a significant decrease in both LT-α and P01375 -α was observed when only good ACR50 responders were analysed . Despite higher levels of baseline synovial P01375 -α in the good responders , neither baseline LT-α nor P01375 -α could predict clinical response after 8 weeks . A decreasing trend of the synovial levels of LT-α was also observed in good responders to infliximab , but the difference did not reach statistical significance . CONCLUSIONS : DB00005 treatment modulates the synovial expression of both P01375 -α and LT-α in vivo , a mechanism that might partly explain its clinical efficacy in RA . Juvenile idiopathic arthritis : will etanercept be an improvement over current therapies ? Overexpression of cytokines in inflamed joints plays an important role in joint inflammation and in damage to articular tissue . Biological agents aimed at specifically antagonising tumour necrosis factor ( P01375 ) are effective in the treatment of adult rheumatoid arthritis . A recent trial of etanercept , a genetically engineered fusion protein consisting of the Fc domain of human IgG1 and the P01375 receptor p75 , has demonstrated that this agent is also well tolerated and effective in patients with juvenile idiopathic arthritis ( JIA ) . DB00005 offers a promising new alternative for patients with JIA who have persistently active arthritis despite treatment with methotrexate . Further studies are needed to clarify whether etanercept is equally effective in the various onset types of JIA ( oligoarthritis , polyarthritis and systemic arthritis ) , whether it can modify disease progression and whether it can be administered safely for long periods of time to children . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . The role of atorvastatin in regulating the immune response leading to vascular damage in a model of Kawasaki disease . Superantigens have been implicated in a number of diseases including Kawasaki disease ( KD ) , a multi-system vasculitis resulting in coronary artery aneurysms . We have characterized a murine disease model in which coronary arteritis is induced by a novel superantigen found in Lactobacillus casei cell wall extract ( LCWE ) . Using this animal model of KD , we have identified three pathogenic steps leading to coronary artery aneurysm formation . These steps include T cell activation and proliferation , production of the proinflammatory cytokine tumour necrosis factor ( P01375 ) -α and up-regulation of matrix metalloproteinase 9 ( P14780 ) , an elastolytic protease . In addition to their cholesterol-lowering effects , 3-hydroxy-3-methylglutaryl ( HMG ) coenzyme A ( DB01992 ) reductase inhibitors ( statins ) have pleotropic immunomodulatory properties . Thus , we examined the effect of atorvastatin in modulating each of these three critical pathogenic processes leading to aneurysm formation in the disease model . DB01076 inhibited lymphocyte proliferation in response to superantigen stimulation in a dose-dependent manner . This inhibition was also observed for production of soluble mediators of inflammation including interleukin ( IL ) -2 and P01375 -α . The inhibitory effect on proliferation was rescued completely by mevalonic acid , confirming that the mechanism responsible for this inhibitory activity on immune activation was inhibition of P04035 . Similarly , P01375 -α-induced P14780 production was reduced in a dose-dependent manner in response to atorvastatin . Inhibition of extracellular-regulated kinase ( P29323 ) phosphorylation appears to be the mechanism responsible for inhibition of P14780 production . In conclusion , atorvastatin is able to inhibit critical steps known to be important in the development of coronary aneurysms , suggesting that statins may have therapeutic benefit in patients with KD . Identification of the fused bicyclic 4-amino-2-phenylpyrimidine derivatives as novel and potent DB05876 inhibitors . 2-Phenyl-4-piperidinyl-6,7-dihydrothieno[3,4-d]pyrimidine derivative ( 2 ) was found to be a new DB05876 inhibitor with moderate Q07343 activity ( IC50=150 nM ) . A number of derivatives with a variety of 4-amino substituents and fused bicyclic pyrimidines were synthesized . Among these , 5,5-dioxo-7,8-dihydro-6H-thiopyrano[3,2-d]pyrimidine derivative ( 18 ) showed potent Q07343 inhibitory activity ( IC50=25 nM ) . Finally , N-propylacetamide derivative ( 31b ) was determined as a potent inhibitor for both Q07343 ( IC50=7.5 nM ) and P01375 -α production in mouse splenocytes ( IC50=9.8 nM ) and showed good in vivo anti-inflammatory activity in the LPS-induced lung inflammation model in mice ( ID50=18 mg/kg ) . The binding mode of the new inhibitor ( 31e ) in the catalytic site of Q07343 is presented based on an X-ray crystal structure of the ligand-enzyme complex . Role of tumour necrosis factor alpha , but not of cyclo-oxygenase-2-derived eicosanoids , on functional and morphological indices of dystrophic progression in mdx mice : a pharmacological approach . The role of tumour necrosis factor ( P01375 ) -alpha or cyclo-oxygenase-2 ( P35354 ) eicosanoids in dystrophinopathies has been evaluated by chronically treating ( 4-8 weeks ) adult dystrophic mdx mice with the anti- P01375 etanercept ( 0.5 mg/kg ) or the P35354 inhibitor meloxicam ( 0.2 mg/kg ) . Throughout the treatment period the mdx mice underwent a protocol of exercise on treadmill in order to worsen the pathology progression ; gastrocnemious muscles from exercised mdx mice showed an intense staining for P01375 by immunohistochemistry . In vivo , etanercept , but not meloxicam , contrasted the exercise-induced forelimb force drop . Electrophysiological recordings ex vivo , showed that etanercept counteracted the decrease in chloride channel function ( gCl ) , a functional index of myofibre damage , in both diaphragm and extensor digitorum longus ( Q9Y5X9 ) muscle , meloxicam being effective only in Q9Y5X9 muscle . None of the drugs ameliorated calcium homeostasis detected by electrophysiology and/or spectrofluorimetry . DB00005 , more than meloxicam , effectively reduced plasma creatine kinase ( CK ) . DB00005 -treated muscles showed a reduction of connective tissue area and of pro-fibrotic cytokine TGF-beta1 vs. untreated ones ; however , the histological profile was weakly ameliorated . In order to better evaluate the impact of etanercept treatment on histology , a 4-week treatment was performed on 2-week-old mdx mice , so to match the first spontaneous degeneration cycle . The histology profile of gastrocnemious was significantly improved with a reduction of degenerating area ; however , CK levels were only slightly lower . The present results support a key role of P01375 , but not of P35354 products , in different phases of dystrophic progression . Anti- P01375 drugs may be useful in combined therapies for Duchenne patients . Differentiating the efficacy of tumor necrosis factor inhibitors . Blockade of tumor necrosis factor ( P01375 ) has emerged as one of the most promising therapies in rheumatoid arthritis ( RA ) . Three agents are currently available as specific P01375 antagonists , etanercept ( Enbrel ) , infliximab ( Remicade ) , and adalimumab ( Humira ) . Data from noncomparative trials suggest that all 3 agents have comparable therapeutic activity in RA . DB00005 and infliximab have also demonstrated beneficial activity in other inflammatory arthritides [ i.e. , psoriatic arthritis and ankylosing spondylitis ( both agents ) and juvenile rheumatoid arthritis ( etanercept only ) ] and inflammatory diseases ( i.e. , psoriasis and uveitis ) . Their effects in granulomatous diseases are more variable , with only infliximab demonstrating clear efficacy in the treatment of Crohn 's disease , sarcoidosis , and Wegener 's vasculitis . In this brief review current efficacy data are summarized and possible explanations for observed clinical differences are explored . [ DB00005 ( Enbrel ) for the treatment of moderate to severe plaque type psoriasis ] . DB00005 ( Enbrel ) is a soluble tumour necrosis factor ( P01375 ) receptor drug . By this mechanism , it inhibits the effects of P01375 involved in the pathogenesis of psoriasis . Enbrel is approved for the treatment of moderate to severe chronic plaque psoriasis , and also for psoriatic arthritis . P01375 and its inhibitors in cancer . P01375 ( P01375 ) -alpha is implicated in the same time in apoptosis and in cell proliferation . P01375 not only acts as pro-inflammatory cytokine conducing to wide spectrum of human diseases including inflammatory diseases , but can also induce tumor development . The molecular mechanisms of P01375 functions have been intensively investigated . In this review we covered P01375 , the molecule , its signaling pathway , and its therapeutic functions . We provide a particular insight in its paradoxical role in tumor promotion and in its use as anti-tumor agent . This review considers also the recent findings regarding P01375 inhibitors , their pharmacokinetics , and their pharmacodynamics . Six P01375 inhibitors have been considered here : DB00065 , DB00051 , Golimumab , DB08904 , CDP571 , DB00005 , and Thalidomide . We discussed the clinical relevance of their functions in treatment of several diseases such as advanced inflammatory rheumatic and bowel disease , with a focus in cancer treatment . Targeting P01375 by these drugs has many side effects like malignancies development , and the long-term sequels are not very well explored . Their efficacy and their safety were discussed , underscoring the necessity of close patients monitoring and of their caution use . [ Biologic therapy with anti- P01375 α in rheumathoid arthritis ] . OBJECTIVE : To evaluate the efficacy , the tolerability , and treatment survival of the association of anti- P01375 α ( DB00065 , DB00005 , DB00051 ) plus DB00563 vs DB00563 as monotherapy in patients with reumathoid arthritis ( RA ) . METHODS : Review of published controlled randomized clinical studies on the association of anti- P01375 α plus DB00563 vs DB00563 alone in patients with rheumathoid arthritis ( RA ) . Results in terms of remission and progression of radiologic damage , and clinical response expressed in terms of P10323 50 or P10323 70 were reviewed in the different studies . RESULTS : In patients with RA the association of anti- P01375 α plus DB00563 led to a greater remission of radiologic damage and marked improvement of clinical response expressed as P10323 50 or P10323 70 compared to therapy with DB00563 only . CONCLUSIONS : In the absence of controlled studies , review of the published studies showed that in RA patients the association of anti- P01375 α plus DB00563 is extremely more efficacious than therapy with DB00563 only . Choice of the drug depends on the activity of the disease , the necessity of a fast response , the presence of side effects , the schedule of treatment and consequently the direct and indirect costs of the drug , and the easiness on supply and distribution . Effects of cytokines on P15692 expression and secretion by human first trimester trophoblast cell line . PROBLEM : The mechanism through which vascular endothelial growth factor ( P15692 ) regulation occurs at the feto-maternal interface is poorly understood . The aim of this study was to investigate the effects of various cytokines on P15692 expression and secretion by trophoblast cells . METHOD OF STUDY : We investigated the effects of cytokines on P15692 expression in human first trimester trophoblast cell line by analyzing P15692 messenger RNA ( mRNA ) by reverse transcription-polymerase chain reaction and P15692 protein secretion by enzyme linked immunosorbent assay . RESULTS : The trophoblast cells expressed P15692 mRNA constitutively and the main subtypes were identified as VEGF121 and VEGF165 . When cultured in the presence of interferon ( IFN ) -gamma , interleukin ( IL ) - 1beta , tumor necrosis factor ( P01375 ) -alpha , P60568 , or P22301 , P15692 mRNA expression was found to be significantly increased by IL-1beta , P01579 and P01375 but to be unaffected by P60568 and P22301 . Moreover , P15692 secretion was most significantly increased by P01579 treatment . CONCLUSION : These results suggest that IL-1beta , P01579 , and P01375 may regulate the production of P15692 in early gestational trophoblasts . Disrupting cytokine signaling in pancreatic cancer : a phase I/II study of etanercept in combination with gemcitabine in patients with advanced disease . OBJECTIVES : DB00005 blocks tumor necrosis factor α ( P01375 -α ) , a proinflammatory cytokine that plays a role in cancer-related cachexia and tumor growth . A phase I/II study was conducted to assess the tolerability and efficacy of gemcitabine and etanercept in advanced pancreatic cancer . METHODS : Twenty-five patients received etanercept 25 mg subcutaneously twice weekly with gemcitabine . A control cohort of 8 patients received gemcitabine alone . The primary end point was progression-free survival at 6 months . Blood specimens were analyzed for P01375 -α , IL-1β , P05231 , interferon-γ , P22301 , and NF-κβ activation . The trial is registered with ClinicalTrials.gov , number NCT00201838 . RESULTS : Thirty-eight patients participated in this study . In the gemcitabine-etanercept cohort , grade 3/4 drug-related toxicities included leucopenia ( 3 ) and neutropenia ( 6 ) . There were 3 ( 12 % ) patients with partial response and 8 ( 32 % ) patients with stable disease . The rate of progression-free survival at 6 months was 28 % [ n = 7 ; 95 % confidence interval ( CI ) , 20 % -36 % ] . Median time to progression was 2.23 months ( 95 % CI , 1.86-4.36 months ) and median overall survival was 5.43 months ( 95 % CI , 3.30-10.23 months ) . Clinical benefit rate was 33 % of the evaluable patients . A correlation was seen between P22301 levels and clinical benefit . CONCLUSIONS : DB00005 added to gemcitabine is safe but did not show significant enhancement of gemcitabine in patients with advanced pancreatic cancer . [ DB00005 : recombinant human soluble tumor necrosis factor receptor fusion protein ] . P01375 is a central cytokine in the pathogenesis of rheumatoid arthritis(RA) , which induces synovitis as well as joint damage . DB00005 is a recombinant human soluble P01375 receptor that binds specifically to P01375 receptor , and inhibits P01375 receptor-mediated signaling cascade . Recent investigations have revealed successful clinical efficacy of biologics in RA including etanercept . We reviewed here the structure and efficacy of etanercept in RA according to the published evidence . DB00005 prevents decrease of cochlear blood flow dose-dependently caused by tumor necrosis factor alpha . OBJECTIVES : P01375 alpha ( P01375 ) is a mediator of inflammation and microcirculation in the cochlea . This study aimed to quantify the effect of a local increase of P01375 and study the effect of its interaction with etanercept on cochlear microcirculation . METHODS : Cochlear lateral wall vessels were exposed surgically and assessed by intravital microscopy in guinea pigs in vivo . First , 24 animals were randomly distributed into 4 groups of 6 each . Exposed vessels were superfused repeatedly either with 1 of 3 different concentrations of P01375 ( 5.0 , 0.5 , and 0.05 ng/mL ) or with placebo ( 0.9 % saline solution ) . Second , 12 animals were randomly distributed into 2 groups of 6 each . Vessels were pretreated with etanercept ( 1.0 microg/ mL ) or placebo ( 0.9 % saline solution ) , and then treated by repeated superfusion with P01375 ( 5.0 ng/mL ) . RESULTS : P01375 was shown to be effective in decreasing cochlear blood flow at a dose of 5.0 ng/mL ( p < 0.01 , analysis of variance on ranks ) . Lower concentrations or placebo treatment did not lead to significant changes . After pretreatment with etanercept , P01375 at a dose of 5.0 ng/mL no longer led to a change in cochlear blood flow . CONCLUSIONS : The decreasing effect that P01375 has on cochlear blood flow is dose-dependent . DB00005 abrogates this effect . DB00005 suppresses arteritis in a murine model of kawasaki disease : a comparative study involving different biological agents . Coronary arteritis , a complication of Kawasaki disease ( KD ) , can be refractory to immunoglobulin ( IVIG ) treatment . To determine the most effective alternative therapy , we compared the efficacy of different agents in a mouse model of KD . Vasculitis was induced by injection of Candida albicans water-soluble fractions ( CAWS ) into a DBA/2 mouse , followed by administration of IVIG , etanercept , methylprednisolone ( MP ) , and cyclosporine-A ( DB00091 ) . At 2 and 4 weeks , the mice were sacrificed , and plasma cytokines and chemokines were measured . CAWS injection induced active inflammation in the aortic root and coronary arteries . At 2 weeks , the vasculitis was reduced only by etanercept , and this effect persisted for the subsequent 2 weeks . At 4 weeks , IVIG and DB00091 also attenuated the inflammation , but the effect of etanercept was more significant . MP exerted no apparent effect at 2 or 4 weeks . The suppressive effect exerted by etanercept on cytokines , such as interleukin- (IL-)6 , IL-12 , P35225 , and tumor necrosis factor- α ( P01375 - α ) , was more evident than that of others . The extent of arteritis correlated with the plasma P01375 - α levels , suggesting a pivotal role of P01375 - α in KD . In conclusion , etanercept was most effective in suppressing CAWS-induced vasculitis and can be a new therapeutic intervention for KD . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . Brain protection from stroke with intravenous TNFα decoy receptor-Trojan horse fusion protein . P01375 ( P01375 ) -α is produced in brain in response to acute cerebral ischemia , and promotes neuronal apoptosis . Biologic P01375 inhibitors ( TNFIs ) , such as the etanercept , can not be developed as new stroke treatments because these large molecule drugs do not cross the blood-brain barrier ( BBB ) . A BBB-penetrating biologic TNFI was engineered by fusion of the type II human P01375 receptor ( TNFR ) to each heavy chain of a genetically engineered chimeric monoclonal antibody ( MAb ) against the mouse transferrin receptor ( P02786 ) , designated as cTfRMAb-TNFR fusion protein . The cTfRMAb domain of the fusion protein acts as a molecular Trojan horse to deliver the fused TNFR across the BBB . DB00005 or the cTfRMAb-TNFR fusion protein ( 1 mg/kg ) was administered intravenously in adult mice subjected to 1-hour reversible middle cerebral artery occlusion up to 90 minutes after the occlusion . Neuroprotection was assessed at 24 hours or 7 days after occlusion . The cTfRMAb-TNFR fusion protein treatment caused a significant 45 % , 48 % , 42 % , and 54 % reduction in hemispheric , cortical , and subcortical stroke volumes , and neural deficit , respectively . Intravenous etanercept had no therapeutic effect . Biologic TNFIs can be reengineered for BBB penetration , and the IgG-TNFR fusion protein is therapeutic after delayed intravenous administration in experimental stroke . Effects of C-phycocyanin and Spirulina on salicylate-induced tinnitus , expression of DB01221 receptor and inflammatory genes . Effects of C-phycocyanin ( C-PC ) , the active component of Spirulina platensis water extract on the expressions of N-methyl D-aspartate receptor subunit 2B ( Q13224 ) , tumor necrosis factor-α ( P01375 -α ) , interleukin-1β ( IL-1β ) , and cyclooxygenase type 2 ( P35354 ) genes in the cochlea and inferior colliculus ( IC ) of mice were evaluated after tinnitus was induced by intraperitoneal injection of salicylate . The results showed that 4-day salicylate treatment ( unlike 4-day saline treatment ) caused a significant increase in Q13224 , P01375 -α , and IL-1β mRNAs expression in the cochlea and IC . On the other hand , dietary supplementation with C-PC or Spirulina platensis water extract significantly reduced the salicylate-induced tinnitus and down-regulated the mRNAs expression of Q13224 , P01375 -α , IL-1β mRNAs , and P35354 genes in the cochlea and IC of mice . The changes of protein expression levels were generally correlated with those of mRNAs expression levels in the IC for above genes . Pharmacogenetics of etanercept in rheumatoid arthritis . DB00005 is one of several P01375 inhibitors approved for rheumatoid arthritis ( RA ) and a variety of other immune-mediated inflammatory conditions . Given the plethora of drugs approved for RA , and the wide variations in cost and treatment response , markers of efficacy would be very useful . Several candidate genes , including HLA- Q8IUH3 alleles and those encoding P01375 , P01375 receptors and Fc receptors , have been examined for a role in the response to treatment with etanercept . In this review , we discuss pharmacogenetic studies of etanercept in RA and other diseases , and comment on the future of such analyses to advance the goal of personalized medicine in RA . DB00005 improves lipid profile and oxidative stress measures in patients with juvenile idiopathic arthritis . OBJECTIVE : To investigate the effect of 1-year treatment with the anti-tumor necrosis factor-α ( P01375 -α ) drug etanercept on lipid profile and oxidative stress in children and adolescents with juvenile idiopathic arthritis ( JIA ) . METHODS : Thirty children with JIA ( 22 females ; mean age 12.3 ± SD 5.7 yrs ) , all eligible for anti- P01375 -α treatment , were assessed at baseline and after 6- and 12-month treatment with etanercept . Disease activity was determined using the Juvenile Arthritis Disease Activity Score ( JADAS ) . Blood samples were drawn to measure the acute-phase reactants P02741 ( CRP ) and erythrocyte sedimentation rate ( P03372 ) , lipids , and the proinflammatory cytokines P01375 -α , interleukin-1β ( IL-1β ) , P05231 and interferon-γ . To measure the oxidative stress marker 8-iso-prostaglandin F2α , 24-h urine samples were collected . RESULTS : Inflammatory indicators ( CRP and P03372 ) and JADAS scores improved significantly after 1 year of etanercept treatment ( all p < 0.001 ) . Proinflammatory cytokines showed significant reduction during the study period ( all p < 0.001 ) . Similar reductions were detected in total cholesterol ( p < 0.001 ) , low-density lipoprotein cholesterol ( p = 0.04 ) , and triglycerides ( p < 0.001 ) , whereas no significant change was found in high-density lipoprotein cholesterol . No side effects were observed during the treatment period . CONCLUSION : This study shows for the first time that anti- P01375 -α therapy for JIA is associated not only with a beneficial effect on clinical disease activity and inflammatory indexes , but also with improved lipid profile and oxidative stress . These findings suggest that P01375 -α blockers might reduce atherosclerotic risk in children with JIA . DB00005 for psoriasis : two case reports . Psoriasis is a chronic inflammatory skin disorder . Recent advances in the understanding of its immunological basis have led to its redefinition as being T-cell mediated . New biological agents have been developed as effective selective target therapies and promise to be an alternative to conventional systemic medications . DB00005 is a recombinant human protein recently approved for psoriatic arthritis treatment that has activity against tumor necrosis factor-alpha ( P01375 ) . It is composed of the human P01375 receptor linked to the Fc portion of human IgG1 . P01375 seems to play a key role in the pathogenesis of psoriasis and psoriatic arthritis . Therefore , etanercept P01375 antagonism is an effective approach for severe psoriasis . We describe two case reports of severe recalcitrant psoriasis , also with arthritis , that showed a remarkable improvement with etanercept , with no adverse events . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . P01375 -induced P16581 expression on vascular endothelial cells . OBJECTIVE : To determine the tumor necrosis factor ( P01375 ) receptor type involved in induction of P16581 expression on vascular endothelial cells . DESIGN : Prospective , in vitro repeated-measures analysis of cellular responses . SETTING : Research laboratory in an academic medical center . SUBJECTS : Cultured human umbilical vein endothelial cells . INTERVENTIONS : Human umbilical vein endothelial cells were incubated with recombinant human P01375 ( rhTNF ) to induce the expression of P16581 on their surfaces . To block rhTNF from binding to receptors , the cells were incubated with monoclonal antibodies against P01375 receptors ( anti-CD120a and anti- DB00005 ) . P01375 -induced P16581 expression of the endothelial cells , with and without blocking antibodies , was then determined using indirect immunofluorescence and flow cytometry . MEASUREMENTS AND MAIN RESULTS : Blocking of either CD120a or DB00005 receptors individually resulted in inhibition of P01375 -induced P16581 expression on human umbilical vein endothelial cells . When both antibodies were added , the inhibition of P01375 -induced P16581 expression was synergistic . Inhibition of P16581 expression was dependent on both P01375 concentrations and antibody concentrations . CONCLUSIONS : Both CD120a and DB00005 receptors are involved in P01375 -induced P16581 expression on human umbilical vein endothelial cells . Blocking of both or one receptor type can reduce or totally inhibit expression of P16581 on human umbilical vein endothelial cells , but the response is dependent on both P01375 and antibody concentrations . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . Expression of tumor necrosis factor-receptor superfamily members by lung T lymphocytes in interstitial lung disease . Recently , a novel receptor superfamily has been identified whose members interact with a parallel family of ligands showing homology to tumor necrosis factor ( P01375 ) . To investigate the role of these receptor structures in the pulmonary environment , we evaluated the expression of some members of the P01375 -receptor ( P26842 , P28908 , P25942 , CD95/Fas , CD120a , and DB00005 ) and P01375 -ligand ( P29965 , P32970 / P32970 , P32971 , and mTNFalpha ) superfamilies by bronchoalveolar lavage ( BAL ) T cells recovered from healthy subjects and patients with interstitial lung disease ( ILD ) . Lung T lymphocytes recovered from control subjects showed a slight expression of P26842 but did not bear P28908 , P25942 , CD120a , or DB00005 antigens . P26842 expression was restricted to normal P01730 + cells . Fas antigen ( CD95 ) , which is involved in activation-driven T-cell suicide , and the ligand for P26842 ( P32970 ) were weakly expressed by normal BAL T-cell subpopulations . In patients with sarcoidosis , the majority of pulmonary T lymphocytes were P01730 + cells that expressed low levels of P26842 and an upregulation of CD95 and P32970 molecules . When we characterized lymphocytes accumulating in the lung of patients with HIV infection and hypersensitivity pneumonitis , we demonstrated that T cells accounting for the CD8 alveolitis bore P01375 -receptor type 2 ( DB00005 ) at high density and were P32970 + while P29965 , P32971 , or mTNF-alpha expression were not found . The discrete surface expression of the P01375 -receptors and P01375 -ligands on alveolar T-cell subsets suggests that these molecules play a role in the immune regulatory mechanisms that ultimately lead to the alveolitis in the pulmonary microenvironment of interstitial lung disease . DB00005 treatment reduces the serum levels of interleukin-15 and interferon-gamma inducible protein-10 in patients with rheumatoid arthritis . P01375 -alpha ( P01375 ) has an essential role in the pathogenesis of rheumatoid arthritis ( RA ) and has been known to induce the production of several inflammatory molecules in vivo . To analyze in vivo the active mechanism of the P01375 blocking agent , etanercept , the serum levels of the cytokine interleukin-15 ( P40933 ) and the chemokines growth-regulated protein-alpha ( Gro-alpha ) , and interferon-gamma inducible protein-10 ( P02778 ) in RA patients were measured . Twenty-two patients with RA were administered etanercept once or twice a week for more than 6 months . The clinical and laboratory parameters were measured and serum levels of P40933 , Gro-alpha , and P02778 were determined using enzyme-linked immunosorbent assay ( ELISA ) kits at the baseline and at 3 and 6 months after the initial treatment . Additionally , the production of P40933 and P02778 by cultured synovial cells stimulated with P01375 from RA patients was determined by ELISA . A significant decrease in serum levels of P40933 and P02778 was observed at 3 and 6 months after initial treatment with etanercept , but not in those of Gro-alpha . P01375 induced production of P02778 , but not P40933 in cultured synovial cells from RA patients . This study demonstrated for the first time the reduction of P02778 and P40933 production in RA patients as active mechanisms of etanercept . Deficient spontaneous in vitro apoptosis and increased tmTNF reverse signaling-induced apoptosis of monocytes predict suboptimal therapeutic response of rheumatoid arthritis to P01375 inhibition . INTRODUCTION : In vitro apoptosis of peripheral monocytes in rheumatoid arthritis ( RA ) is disturbed and influenced by cytokine production and transmembrane P01375 ( tmTNF ) reverse signaling . The goal of the study was the analysis of the predictive value of the rate of in vitro apoptosis for the therapeutic response to anti- P01375 treatment . METHODS : Spontaneous and tmTNF reverse signaling-induced apoptosis were determined in vitro in monocytes from 20 RA patients prior to initiation of therapeutic P01375 inhibition with etanercept , and the subsequent clinical response was monitored . RESULTS : Spontaneous in vitro apoptosis was significantly reduced in RA patients compared to controls . Deficiency in spontaneous apoptosis was associated with an insufficient therapeutic response according to the European League Against Rheumatism ( EULAR ) response criteria and less reduction of the disease activity determined by disease activity score ( DAS ) 28 . High susceptibility to reverse signaling-induced apoptosis was also associated with less efficient reduction in the DAS28 . Of note , a strong negative correlation between the two apoptotic parameters was discernible , possibly indicative of two pathogenetically relevant processes counter-regulating each other . tmTNF reverse signaling induced in vitro production of soluble IL1-RI and IL-1RII only in monocytes not deficient in spontaneous apoptosis , and the levels of soluble IL1-RII were found to be predictive of a good clinical response to DB00005 . CONCLUSION : Although tmTNF reverse signaling is able to induce apoptosis of RA monocytes in vitro , this process appears to occur in vitro preferentially in patients with suboptimal therapeutic response . Resistance to spontaneous in vitro apoptosis , in contrast , is a predictor of insufficient response to treatment . Microglial activation , increased P01375 and P31645 expression in the prefrontal cortex define stress-altered behaviour in mice susceptible to anhedonia . A chronic stress paradigm comprising exposure to predation , tail suspension and restraint induces a depressive syndrome in C57BL/6J mice that occurs in some , but not all , animals . Here , we sought to extend our behavioural studies to investigate how susceptibility ( sucrose preference < 65 % ) or resilience ( sucrose preference > 65 % ) to stress-induced anhedonia affects the 5HT system and the expression of inflammation-related genes . All chronically stressed animals , displayed increased level of anxiety , but susceptible mice exhibited an increased propensity to float in the forced swim test and demonstrate hyperactivity under stressful lighting conditions . These changes were not present in resilient or acutely stressed animals . Compared to resilient animals , susceptible mice showed elevated expression of tumour necrosis factor alpha ( P01375 ) and the 5-HT transporter ( P31645 ) in the pre-frontal area . Enhanced expression of 5HT(2A) and P23219 in the pre-frontal area was observed in all stressed animals . In turn , indoleamine-2,3-dioxygenase ( P14902 ) was significantly unregulated in the raphe of susceptible animals . At the cellular level , increased numbers of Iba-1-positive microglial cells were also present in the prefrontal area of susceptible animals compared to resilient animals . Consequently , the susceptible animals display a unique molecular profile when compared to resilient , but anxious , animals . Unexpectedly , this altered profile provides a rationale for exploring anti-inflammatory , and possibly , P01375 -targeted therapy for major depression . [ Cytokines -- causes , players or bystanders in heart failure ] . BACKGROUND : Cytokines are important mediators of the immune system and are of pathophysiological relevance for different cardiac diseases . Currently , 18 cytokines carrying the name interleukin ( IL ) are known ; they can be subdivided into pro- and anti-inflammatory interleukins . CARDIAC CYTOKINES : They partly act in a negative inotropic manner and cause destruction of cardiomyocytes resulting in myocardial fibrosis . The proinflammatory cytokine tumor necrosis factor ( P01375 -)alpha induces cardiodepressive effects and causes apoptosis . P01375 , P05231 , soluble P01375 -receptor-1 and -2 are independent predictors of increased mortality of patients with heart failure . Experimental and clinical evidence has shown that plasma and tissue levels of P01375 were elevated to such extent as to explain at least some of the symptoms of heart failure due to the actions of this cytokine . CLINICAL TRIALS : Two multicenter studies ( RENAISSANCE , RECOVER ) , based on promising pilot studies , have disclosed no effect for the P01375 antagonists ( DB00005 ) on mortality and morbidity . It is not possible , however , to draw the conclusion from the data of these studies that P01375 plays no significant pathophysiological role in the etiology and progression of heart failure . Effect of P35354 inhibitor lumiracoxib and the P01375 -α antagonist etanercept on TNBS-induced colitis in Wistar rats . Crohn 's disease ( CD ) is associated with gut barrier dysfunction . Besides the baseline barrier defect , a subgroup of patients also expresses an intestinal barrier hyperresponsiveness to nonsteroidal anti-inflammatory drugs . On the other hand , the anti-tumour necrosis factor alpha ( P01375 -α ) treatment has brought benefits to these patients . Thus , this study aimed to evaluate the effect of lumiracoxib , a selective-cyclooxygenase-2 ( P35354 ) inhibitor , and DB00005 ( ETC ) , a P01375 -α antagonist on the 2,4,6-trinitrobenzene sulfonic acid ( TNBS ) -induced experimental colitis . A total of 47 Wistar rats were randomized into seven groups , as follows : ( 1 ) Sham : sham induced-colitis ; ( 2 ) TNBS : nontreated induced-colitis ; ( 3 ) DB01283 control ; ( 4 ) DB01283 -treated induced-colitis ; ( 5 ) ETC control ; ( 6 ) ETC-treated induced-colitis ; ( 7 ) DB01283 -ETC-treated induced-colitis . Rats from groups 6 and 7 presented significant improvement of macroscopic and histopathological damages in the distal colon . The gene expression of P35354 mRNA , as well of P01375 -α mRNA , decreased significantly in groups 6 and 7 compared to the TNBS nontreated and lumiracoxib-treated groups . The treatment only with lumiracoxib did not reduce the inflammation on TNBS-induced experimental colitis . ETC attenuated the damage seen in the colon and reduced the inflammation caused by TNBS . Our results suggest that down-regulation of P01375 -α and P35354 resulted in a decrease in inflammation caused by TNBS and thus provided some protection from the colonic damage caused by TNBS . Clastogenic plasma factors in psoriasis -- comparison of phototherapy and anti- P01375 -α treatments . As previously described , Psoralen plus UVA ( PUVA ) therapy induces chromosome damage in psoriatic patients . This study evaluates whether these effects are transitory or persistent . In addition , we studied these effects after narrowband UVB ( nUVB ) and anti-tumor necrosis factor ( P01375 ) -α treatments . Among 40 responder patients , 10 received PUVA , 10 nUVB , 10 DB00065 and 10 DB00005 . Disease activity was determined with Psoriasis Area and Severity Index . Chromosomal breakage was evaluated by the clastogenic factor ( CF ) test . Potential clastogenic agents , malondialdehyde ( MDA ) and P01375 -α were measured . Before treatment , the plasma-adjusted clastogenic scores ( ACS ) of patients were increased . During treatment , a further increase in ACS was observed in both phototherapy groups . Chromosome damage persisted for PUVA patients at week 32 , while it diminished after nUVB to ACS values lower than before treatment . MDA and P01375 -α values were also increased at baseline . MDA decreased during treatment in all groups , but without reaching normal levels . Plasma P01375 -α remained unchanged in PUVA and nUVB but decreased in both anti- P01375 -α treatment groups . Psoriasis is accompanied by CF-induced chromosomal breakage that increases during PUVA and nUVB treatments . Plasma clastogenic activity persisted in the follow-up after PUVA , while after nUVB ACS returned to values even lower than baseline . Clastogenic activity during the induction phase with anti- P01375 -α remained unchanged . Expression of P01375 receptors by T cells and membrane P01375 by alveolar macrophages suggests a role for P01375 in the regulation of the local immune responses in the lung of HIV-1-infected patients . High amounts of P01375 are released by alveolar macrophages ( AMs ) in the lungs of patients with HIV-1 infection . To investigate the role of this cytokine in the local immune response , we studied the expression of surface receptors for P01375 ( P01375 -Rs ) and the presence of the transmembrane form of P01375 ( mTNF-alpha ) on bronchoalveolar lavage ( BAL ) cells recovered from 14 patients with HIV-1 infection . The role of P01375 both in the events leading to the T cell alveolitis and as a mediator of cytotoxicity was also evaluated . P01375 -R expression was determined by flow cytometry on BAL CD8 lymphocytes and AMs ( i.e. , the cells that account for the alveolitis in HIV-1 infection ) . We found that CD8 cells express the 75-kDa ( CD120a ) but not the 55-kDa ( CD120a ) P01375 -Rs , whereas AMs were devoid of P01375 -R expression . More than 90 % of BAL T cells efficiently bound P01375 ; when T cells were tested for their proliferative capacity , an up-regulation of the P60568 -mediated proliferation by P01375 was observed , suggesting that this cytokine may drive the in situ proliferation of DB00005 + T cells . As shown by flow cytometry analysis and immunoprecipitation with anti- P01375 Ab , mTNF-alpha expression was observed on AMs but not on alveolar T cells . Fixed AMs showed high levels of killing against P01375 -sensitive targets . Taken together , our data demonstrate the selective expression of P01375 -Rs and mTNF-alpha on cells accumulating within the alveolar spaces of patients with HIV-1 infection , pointing to the compound role of P01375 in the local immune responses . A case of tuberculous arthritis following the use of etanercept . DB00005 is a tumor necrosis factor ( P01375 ) inhibitor that has been used for the treatment of chronic inflammatory diseases including rheumatoid arthritis , ankylosing spondylitis and psoriatic arthritis . Because of its immunosuppressive activity , opportunistic infections have been noted in treated patients , most notably caused by Mycobacterium tuberculosis . Tuberculosis may present in an extrapulmonary or disseminated form . Since P01375 inhibitors have been used in Korea , a few cases of P01375 inhibitor associated tuberculosis have been described . However , tuberculous arthritis has not been previously reported . We describe a case of tuberculous arthritis in a 57-year-old woman with rheumatoid arthritis who was treated with etanercept . DB00588 -induced regulation of the balance within macrophage subpopulations . In asthma , treatment with inhaled corticosteroids reduces chronic peribronchial inflammation and restores the balance within macrophage subpopulations . This study investigates whether corticosteroids can regulate monocyte differentiation in vitro and thereby influence the balance of functionally distinct macrophages . Graded doses of fluticasone propionate ( FP ) were added to cultures of normal peripheral blood monocytes in the presence or absence of P05112 . Cells were harvested after 7 days ' culture . Double immunofluorescence studies were performed on cytospins of differentiated macrophages using the MoAbs RFD1 and RFD7 to distinguish inductive and suppressive macrophages by their respective phenotypes . Macrophage function was determined by quantifying allostimulation in a mixed leucocyte reaction and by measuring tumour necrosis factor-alpha ( P01375 ) production . FP reduced the number of mature cells with a D1+ antigen-presenting phenotype and up-regulated the development of cells with the D1/D7+ and D7+ phenotypes . Functionally , this was associated with reduced stimulation of T cell proliferation in a mixed leucocyte reaction ( P08235 ) . DB00588 also reversed the increase in both D1+ expression and P01375 production induced by P05112 . The effect of FP persisted for 24 h after removal of FP from the culture medium . These results suggest that FP treatment of asthmatics may have a direct beneficial effect by normalizing the macrophage subset imbalance that contributes to the chronic peribronchial inflammation present in this condition .

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P10275 stimulates bone sialoprotein ( BSP ) gene transcription via DB02527 response element and activator protein 1/glucocorticoid response elements . Bone sialoprotein ( BSP ) is an early marker of osteoblast differentiation . Androgens are steroid hormones that are essential for skeletal development . The androgen receptor ( AR ) is a transcription factor and a member of the steroid receptor superfamily that plays an important role in male sexual differentiation and prostate cell proliferation . To determine the molecular mechanism involved in the stimulation of bone formation , we have analyzed the effects of androgens and AR effects on BSP gene transcription . AR protein levels were increased after AR overexpression in ROS17/2.8 cells . BSP mRNA levels were increased by AR overexpression . However , the endogenous and overexpressed BSP mRNA levels were not changed by DB02901 ( 10(-8) M , 24 h ) . Whereas luciferase ( LUC ) activities in all constructs , including a short construct ( nts -116 to +60 ) , were increased by AR overexpression , the basal and LUC activities enhanced by AR overexpression were not induced by DB02901 ( 10(-8)M , 24 h ) . The effect of AR overexpression was abrogated by 2 bp mutations in either the DB02527 response element ( CRE ) or activator protein 1 ( P05412 ) /glucocorticoid response element ( GRE ) . Gel shift analyses showed that AR overexpression increased binding to the CRE and P05412 /GRE elements . Notably , the CRE-protein complexes were supershifted by phospho-CREB antibody , and CREB , c-Fos , c-Jun , and AR antibodies disrupted the complexes formation . The P05412 /GRE-protein complexes were supershifted by c-Fos antibody and c-Jun , and AR antibodies disrupted the complexes formation . These studies demonstrate that AR stimulates BSP gene transcription by targeting the CRE and P05412 /GRE elements in the promoter of the rat BSP gene . Hyperresponsive TH2 cells with enhanced nuclear factor-kappa B activation induce atopic dermatitis-like skin lesions in Nishiki-nezumi Cinnamon/Nagoya mice . BACKGROUND : Nishiki-nezumi Cinnamon/Nagoya ( NC/Nga ) mice raised in nonair-controlled conventional circumstances spontaneously develop atopic dermatitis-like skin lesions ; however , the underlying mechanisms remain unclear . OBJECTIVE : We wanted to identify the critical intracellular signaling molecules in T cells that induce atopic dermatitis-like skin legions in NC/Nga mice . METHODS : We examined the levels of signal transduction and cytokine production in T cells , particularly those in atopic dermatitis-like lesions induced by the topical injection of mite antigens in NC/Nga mice under specific pathogen-free conditions . RESULTS : In NC/Nga mice maintained under specific pathogen-free conditions , the capability of T(H)1/T(H)2 and T cytotoxic 1/T cytotoxic 2 ( Tc1/Tc2 ) cell differentiation increased significantly . T-cell antigen receptor-mediated activation of the extracellular signal-regulated kinase/mitogen-activated protein kinase cascade and nuclear factor-kappaB ( NF-kappaB ) signaling were enhanced in NC/Nga T cells . The expression of T(H)2 cytokines ( P05112 , P35225 , and P05113 ) and that of GATA-binding protein 3 ( P23771 ) , avian musculoaponeurotic fibrosarcoma ( c-Maf ) , NF-kappaB , and activator protein 1 ( P05412 ) selectively increased in draining lymph node T cells of NC/Nga mice . Moreover , the cell transfer of inhibitory NF-kappaB mutant-infected T(H)2 cells reduced ear thickness in the mite antigen-induced skin lesion of NC/Nga mice . CONCLUSION : Hyperresponsive T(H)2 cells with an enhanced activity of NF-kappaB and P05412 play a crucial role in the pathogenesis of atopic dermatitis-like skin lesions in NC/Nga mice . CLINICAL IMPLICATIONS : These results indicate potential therapeutic usefulness of developing selective inhibitors for NF-kappaB in the treatment of human atopic dermatitis . Extracellular DB00171 and nerve growth factor intensify hypoglycemia-induced cell death in primary neurons : role of P2 and NGFRp75 receptors . In this study , we monitored the direct expression of P2 receptors for extracellular DB00171 in cerebellar granule neurons undergoing metabolism impairment . DB09341 deprivation for 30-60 min inhibited P47900 receptor protein , only weakly modulated P51575 , Q9UBL9 and P56373 , and up-regulated by about two-fold Q99571 , Q99572 and P51582 . The P2X/Y antagonist basilen blue , protecting cerebellar neurons from hypoglycemic cell death , maintained within basal levels only the expression of Q99572 and P51582 proteins , but not Q99571 or P47900 . DB09341 starvation transiently increased ( up to three-fold ) the expression of NGFRp75 receptor protein and strongly stimulated the extracellular release of nerve growth factor ( P01138 ; about 10-fold ) . Exogenously added P01138 then augmented hypoglycemic neuronal death by about 60 % , increasing the percentage of Höechst-positive nuclei ( from approximately 62 to 95 % ) , reducing lactate dehydrogenase ( LDH ) release ( from about 50 to 14 % ) and significantly overstimulating the hypoglycemia-induced expression of Q99572 and P51582 . Conversely , extracellular DB00171 augmented hypoglycemic neuronal death by about 80 % , reducing the number of Höechst-positive nuclei ( from approximately 62 % to 14 % ) , augmenting LDH outflow ( by about 30 % ) and further increasing the hypoglycemia-induced expression of NGFRp75 . Our results indicate that P2 and NGFRp75 receptors are modulated during glucose starvation and that extracellular DB00171 and P01138 drive features of , respectively , necrotic and apoptotic hypoglycemic cell death , aggravating the consequences of metabolism impairment in cerebellar primary neurons . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . DB01407 affects the expression of messenger RNA for interleukin 10 in peripheral leukocytes from horses challenged intrabronchially with lipopolysaccharides . On four occasions , four horses with heaves and four horses with small airway inflammatory diseases inhaled 0.9 % saline based aerosol mixtures with or without lipopolysaccharides ( LPS ) . Prior to the first saline and LPS inhalation , horses were untreated , while three and a half days prior to the third and forth inhalation horses had received 0.8 microg/kg clenbuterol intravenously twice daily . The messenger RNA ( mRNA ) expression of tumour necrosis factor-alpha ( P01375 ) , interleukin ( IL ) -1beta , P05112 , P05231 , P10145 , P22301 and interferon- gamma ( IFN- gamma ) was investigated by RT-PCR , all of which were expressed in the white blood cells of samples collected . Inhalation of LPS only changed the cytokine expression profile of P22301 , P05112 and P01375 mRNA which were higher after challenge with LPS . However in those horses that were treated with clenbuterol the LPS-induced P22301 mRNA expression was shown to be suppressed . Further changes in P05112 and P01375 were not significant . Thus the results of this study indicated that clenbuterol can modulate the expression of P22301 mRNA in peripheral white blood cells in those horses with small airway diseases that have been exposed to LPS . Cardiovascular effects of sildenafil in hypertensive men with erectile dysfunction and different alleles of the type 5 cGMP-specific phosphodiesterase ( O76074 ) . Erectile dysfunction ( ED ) is frequent in patients with essential hypertension ( EH ) ; a likely common pathogenetic pathway could be a reduced ability of arteriolar vascular smooth muscle ( VSM ) to relax . Increasing intracellular levels of cGMP reduce the contractile status of VSM ; on the contrary , type 5 cGMP-specific phosphodiesterase ( O76074 , codified by O76074 gene ) regulates cGMP levels through its clearance . The O76074 gene represents a good candidate for the intermediate phenotype EH/ED : genetic variants of the O76074 may predispose to EH and ED and could affect the local and systemic response to sildenafil administration . Thus , a functionally relevant portion of O76074 5'-flanking promoter region was analyzed by PCR and direct sequencing in patients with EH and idiopathic ED . The sequences obtained showed a T/G polymorphism at position -1142 , near an P05412 regulatory element , that was not apparently associated with the intermediate phenotype . We also studied the relationship between this polymorphism and the effects of oral sildenafil on blood pressure ( BP ) and heart rate ( HR ) in men with ED . DB00203 caused a significant decrease of BP , but had no effects on HR ; statistical analysis showed no differences in BP and HR variations among O76074 genotypes . In conclusion , our data showed no correlations of a novel polymorphism of the O76074 promoter gene with the intermediate phenotype EH/ED and the BP and HR response to sildenafil administration . Further studies are necessary to define the role of this polymorphism and to study the genetic predisposition for EH with ED . P10275 expression induces P09038 , FGF-binding protein production , and P09038 release in prostate carcinoma cells : role of P09038 in growth , survival , and androgen receptor down-modulation . BACKGROUND : Alterations in fibroblast growth factors ( FGFs ) production and/or FGF receptors expression have been described to play key roles in prostate tumor progression , particularly in androgen-independent tumors . However , the role of androgen receptor ( AR ) in altering FGF-mediated growth and survival of prostatic neoplastic cells has not been completely defined . In this study , we investigated the alterations in P09038 production and utilization by the PC3 cell line , after transfection with a full-length AR . METHODS : P05230 ,2,7 , FGF-binding protein ( Q14512 ) production and FGF receptor ( FGFR ) 1-4 expression were investigated by polymerase chain reaction ( PCR ) and Western blot analysis . RESULTS : De novo AR expression by PC3 cells restores P21802 IIIb isoform expression and sensitivity to P21781 and P09038 . Androgen stimulation induces AR+ PC3 clones to secrete Q14512 , likely responsible for activation and mobilization from the extracellular matrix of the high amounts of P09038 produced by the same cells . In addition to the effects on cell proliferation , P09038 maintains the survival of AR+ PC3 clones through a positive modulation of the Bcl-2 protein and down-modulates AR protein expression , allowing the escape of selected clones from androgen regulation . CONCLUSION : In the presence of an active AR , the combined production of P09038 and Q14512 may play an important role in the progression of prostate cancer through the selection of AR- clones expressing high levels of Bcl-2 . Differential radiosensitisation by ZD1839 ( DB00317 ) , a highly selective epidermal growth factor receptor tyrosine kinase inhibitor in two related bladder cancer cell lines . The epidermal growth factor receptor ( P00533 ) is expressed in a wide variety of epithelial tumours including carcinoma of the bladder . Stimulation of the P00533 pathway is blocked by ZD1839 ( DB00317 ) , a highly selective P00533 tyrosine kinase inhibitor . Radical radiotherapy is an established organ sparing treatment option for muscle invasive bladder cancer and this study has explored the possibility for the use of ZD1839 as a radiosensitiser in this scenario . The effect of combination treatment with ZD1839 ( 0.01 microM ) and ionising radiation in the established bladder cancer cell lines MGH-U1 and its radiosensitive mutant clone S40b was measured by clonogenic assays . A highly significant radiosensitising effect was seen in both cell lines ( P < 0.001 for MGH-U1 and S40b cell lines ) . This effect was independent of the concentration of the drug and the duration of exposure prior to treatment with ionising radiation . Cell cycle kinetics of both cell lines was not significantly altered with ZD1839 ( 0.01 microM ) as a single agent . A modest induction of apoptosis was observed with ZD1839 ( 0.01 microM ) as a single agent , but a marked induction was observed with the combination treatment of ZD1839 and ionising radiation . These results suggest a potentially important role for ZD1839 in combination with radiotherapy in the treatment of muscle invasive bladder cancer . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . DB01407 induces growth factor mRNA , activates astrocytes , and protects rat brain tissue against ischemic damage . The induction of growth factor synthesis in brain tissue by beta2-adrenoceptor agonists , such as clenbuterol , is a promising approach to protect brain tissue from ischemic damage . DB01407 ( 0.01-0.5 mg/kg ) reduced the cortical infarct volume in Long-Evans rats as measured 7 days after permanent occlusion of the middle cerebral artery . Dosages of clenbuterol higher than 1 mg/kg showed no cerebroprotective effect due to a decrease in blood pressure and an increase in plasma glucose level . The increase in the mRNA level of nerve growth factor ( P01138 ) , basic fibroblast growth factor ( basic FGF ) , and transforming growth factor-beta1 ( TGF-beta1 ) mRNA in cortical and hippocampal tissue occurred earlier after middle cerebral artery occlusion and was more pronounced in animals treated with clenbuterol than in controls . In addition , glial fibrillary acidic protein ( P14136 ) mRNA expression was enhanced in astrocytes 6 h after ischemia in clenbuterol-treated animals . The results suggest that growth factor synthesis is enhanced in activated astrocytes and that this could be the mechanism of clenbuterol-induced cerebroprotection after ischemia . The role of the sympathetic efferents in endotoxin-induced localized inflammatory hyperalgesia and cytokine upregulation . The sympathetic system ( SNS ) is considered to be a major component of the neurogenic contribution to inflammation and hyperalgesia . We have investigated the role of the SNS in the local inflammatory pain induced by intraplantar ( i.pl ) injections of bacterial endotoxin ( ET ) . Treatment of rats with an alpha-adrenoceptor antagonist ( phentolamine , 0.25-1 mg/kg , i.p. ) , a beta-adrenoceptor antagonist ( propranolol , 1-10 mg/kg , p.o. ) or a sympathetic neuron-blocking agent ( guanethedine , 30 mg/kg , s.c. ) resulted in a dose-dependent reduction of the thermal hyperalgesia induced by ET . Mechanical hyperalgesia , however , was less sensitive to inhibition by propranolol and guanethedine but significantly inhibited by phentolamine . ET injection produced significant upregulation of tumor necrosis factor-alpha ( P01375 ) , interleukin-1 beta ( P01584 ) , P05231 , and nerve growth factor ( P01138 ) . Treatment with any one of the three sympatholytics abolished the upregulation of P01138 and P05231 , while phentolamine and guanethedine also reversed the upregulation of P01375 . P01584 was resistant to all of the sympatholytic treatments . We conclude that the SNS can contribute to the local inflammation and hyperalgesia following injection of ET . The resistance to sympatholytics shown by P01584 , known to play a key role in the inflammatory cascade , suggests that ET can initiate inflammation and hyperalgesia independently of peripheral and central sympathetic mechanisms . Effect of interferon-gamma and P01375 on P15941 mucin expression in ovarian carcinoma cell lines . In view of the potential uses of cell surface tumour associated antigens in novel anticancer treatment , a study was designed to investigate whether the biological response modifiers interferon-gamma ( P01579 ) and tumour necrosis factor-alpha ( P01375 ) could effect the expression of an epitope on the tumour associated P15941 epithelial mucin . Four ovarian carcinoma cell lines showing high ( OAW42 and GG ) and low ( JAM and PE01 ) basal expression of P15941 were treated with 10-1000 U/mL of P01579 or P01375 for one or five days . Changes in P15941 expression in cells exposed to P01579 or P01375 were monitored using an ELISA technique with the monoclonal antibody O43633 which reacts with a core protein epitope on the P15941 mucin , and then corrected for the number of viable cells present . P01375 had little effect on P15941 expression , but one or five days exposure to P01579 significantly increased P15941 expression ( p < 0.01 ) in all cell lines including the two cell lines that initially showed little or no expression . [ DB01407 in amyotrophic lateral sclerosis. No indication for a positive effect ] . The anabolic effects of clenbuterol have been recognized for a long time . DB01407 augments the expression of specific muscle proteins with a differential effect on type I and type II fibres . Furthermore , clenbuterol induces the synthesis of endogenous nerve growth factor ( P01138 ) and may itself be a myotrophic factor released by neuron endings . Side effects include tremor and headache and dose dependent abnormalities of laboratory values ( hypokalemia , hypoglycemia ) . After long-term medication increasing fatigue of muscles has been observed . Decreased expression of beta 2-adrenergic receptors may limit the expected functional improvement . The efficacy of clenbuterol as symptomatic treatment of amyotrophic lateral sclerosis has not been proved . Controlled treatment trials are warranted to assess this question . Endotoxin-induced liver damage in rats is minimized by beta 2-adrenoceptor stimulation . OBJECTIVE AND DESIGN : To investigate the effects of beta(2)-adrenoceptor ( beta(2)-AR ) stimulation on endotoxin-induced liver damage and systemic cytokine levels in rats . SUBJECTS : Standard male Wistar rats . TREATMENT : A disease-model of lipopolysaccharide ( LPS ) -induced acute systemic inflammation was used . The beta(2)-selective AR agonist clenbuterol was administered before , during , and after LPS-challenge to investigate its effects on the acute inflammatory response and associated liver-failure . METHODS : The following parameters have been measured in plasma : P01375 alpha , P01584 , P05231 , P22301 , Q9NRA2 , ALT , and Bilirubin . Liver histological examination was performed to look for changes in tissue morphology . RESULTS : Administration of clenbuterol ( p.o. ) one hour before , or intravenous at the same time as LPS-challenge resulted in a marked reduction of plasma levels of P01375 alpha , P01584 , and P05231 . A change both in plasma-level and in time-concentration profile of the anti-inflammatory cytokine P22301 was found . DB01407 minimized LPS-induced liver damage , as represented by significantly lowered concentrations of several parameters for liver-failure ( Q9NRA2 , ALT , Bilirubin ) , and improved hepatic tissue morphology . DB01407 administration after LPS challenge failed to inhibit P01375 alpha-release but reduced liver-damage . Simultaneous use of the beta(2)-AR antagonist propranolol augmented LPS-induced liver failure , suggesting a role of endogenous adrenoceptor-agonists in prevention of organ-failure during systemic inflammation . CONCLUSIONS : The results indicate that a selective beta(2)-AR agonist might be used as an additional therapeutic agent in the clinic for the treatment of ( acute ) systemic inflammatory disorders in order to reduce or prevent subsequent liver failure . The molecular biology of interleukin 6 and its receptor . Functional pleiotropy and redundancy are characteristic features of cytokines . Interleukin 6 ( P05231 ) is a typical example : P05231 induces cellular differentiation or expression of tissue-specific genes ; it is involved in processes such as antibody production in B cells , acute-phase protein synthesis in hepatocytes , megakaryocyte maturation , cytotoxic T cell differentiation , and neural differentiation of PC12 ( pheochromocytoma ) cells . It promotes growth of myeloma/plasmacytoma cells , T cells , keratinocytes and renal mesangial cells , and it inhibits growth of myeloid leukaemic cell lines and certain carcinoma cell lines . The P05231 receptor consists of two polypeptide chains , a ligand-binding chain ( IL-6R ) and a non-ligand-binding , signal-transducing chain ( P40189 ) . Interaction of P05231 with IL-6R triggers the association of P40189 and IL-6R , and the signal can be transduced through P40189 . Association of P40189 with IL-6R is involved in the formation of high affinity binding sites . This two-chain model has been shown to be applicable to receptor systems for several other cytokines , such as granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) , P08700 , P05113 and nerve growth factor ( P01138 ) . The pleiotropy and redundancy of cytokines may be explained on the basis of this unique receptor system . Francisella tularensis alters human neutrophil gene expression : insights into the molecular basis of delayed neutrophil apoptosis . We demonstrated recently that Francisella tularensis profoundly impairs human neutrophil apoptosis , but how this is achieved is largely unknown . Herein we used human oligonucleotide microarrays to test the hypothesis that changes in neutrophil gene expression contribute to this phenotype , and now demonstrate that F. tularensis live vaccine strain ( LVS ) caused significant changes in neutrophil gene expression over a 24-hour time period relative to the uninfected controls . Of approximately 47,000 genes analyzed , 3,435 were significantly up- or downregulated by LVS , including 365 unique genes associated with apoptosis and cell survival . Specific targets in this category included genes asso-ciated with the intrinsic and extrinsic apoptotic pathways ( O15519 , P21580 , Q9UBN6 , P04179 , Q16548 , P98170 , Q9P1W9 , P50591 , O14798 , P42575 and Q14790 ) and genes that act via the NFĸB pathway and other mechanisms to prolong cell viability ( P19838 , Q00653 and Q04206 , P01584 , CAST , P24941 , O75293 , P20749 , Q13489 , P24941 , P01583 , P43490 , P05231 , P09341 , P13236 and P15692 ) . The microarray data were confirmed by qPCR and pathway analysis . Moreover , we demonstrate that the P98170 remained abundant in polymorphonuclear leukocytes over 48 h of LVS infection , whereas Q07812 mRNA and protein were progressively downregulated . These data strongly suggest that antiapoptotic and prosurvival mechanisms collaborate to sustain the viability of F. tularensis -- infected neutrophils . Evaluation of cytokine production by equine alveolar macrophages exposed to lipopolysaccharide , Aspergillus fumigatus , and a suspension of hay dust . OBJECTIVE : To evaluate cytokine production by equine alveolar macrophages after exposure to lipopolysaccharide ( LPS ) , Aspergillus fumigatus , and hay dust , and determine the effect of clenbuterol on the cytokine response . ANIMALS : 6 horses . PROCEDURE : Alveolar macrophages were exposed to PBS solution ( negative control ) , LPS , hyphae and conidia of Aspergillus fumigatus ( AF ) , or a suspension of hay dust ( Q86SQ9 ) and incubated for 24 hours at 37 degrees C . Concentrations of tumor necrosis factor ( P01375 ) -alpha and interleukin ( IL ) -1beta were measured in the supernatant . The procedure was repeated with cells that were concurrently incubated with 0.5 microM clenbuterol . RESULTS : Exposure to Q86SQ9 and AF significantly increased production of P01375 by equine alveolar macrophages . The increase in P01375 produced in response to Q86SQ9 and AF was 5 and 7 times as great , respectively , as the increase measured in response to LPS . The concentration of IL-1beta in the supernatant was significantly increased after exposure of cells to AF . DB01407 was effective at inhibiting P01375 production by cells exposed to LPS , Q86SQ9 , or AF . CONCLUSIONS AND CLINICAL RELEVANCE : Increased production of P01375 and IL-1 indicated that the pro-inflammatory cytokines produced by alveolar macrophages in response to allergens may play a role in recurrent airway obstruction ( O75106 ) in horses . Equine alveolar macrophages are not only a primary pulmonary defense mechanism but may also influence the pathogenesis of equine O75106 . The beta2-adrenoceptor agonist clenbuterol , a drug that is commonly used for treatment of equine O75106 , promotes immediate bronchodilation and may also contribute to downward modulation of the inflammatory response . Influence of polymorphisms and P01375 and IL1β serum concentration on the infliximab response in Crohn 's disease and ulcerative colitis . AIM : Inflammatory bowel diseases ( Q9UKU7 ) , such as Crohn 's disease ( CD ) and ulcerative colitis ( UC ) , are partially attributable to an increased secretion of proinflamatory cytokines , such as tumour necrosis factor ( P01375 ) and interleukin-1β ( IL1β ) , which play essential roles in the disease pathogenesis and are target molecules for specific therapy . Given the inter-individual variability in the response to the anti- P01375 monoclonal antibody infliximab , the aim of our study was to explore the predictive value of P01375 and/or IL1β as surrogate markers of infliximab response . METHODS : Serial serum concentrations of P01375 and IL1β and P01375 promoter region and P01584 polymorphisms were determined in 47 patients ( 29 CD and 18 UC ) receiving infliximab and correlated with treatment response . RESULTS : Baseline serum concentrations of P01375 and IL1β were higher in UC patients than in CD patients ( p = 0.0097 and 0.0024 , respectively ) . CD patients showing < 0.64 pg/ml IL1β at baseline were more frequently responders than non-responders ( p = 0.036 ) , and the C allele of the P01584 polymorphism was associated with higher IL1β serum concentrations ( p = 0.026 ) and with poorer clinical remission after 14 weeks of infliximab treatment . No significant association was found between serum P01375 concentration or P01375 polymorphism and patient response to infliximab . CONCLUSION : This is the first study evaluating the pharmacogenetic role of the rs1143634 polymorphism of P01584 and P01375 polymorphisms in infliximab-treated Q9UKU7 patients . We found an association between the rs1143634 C allele and higher serum IL1β concentrations and a lower response to infliximab treatment in CD patients that warrants the interest of future studies in larger and independent series . DNA polymorphism in cytokine genes based on length variation in simple-sequence tandem repeats . The possibility of the involvement of cytokines in the genetic predisposition to various diseases has been suggested by a large variety of studies . However , the study of potential disease linkage of cytokine genes has been hampered by a lack of sufficiently polymorphic markers at the restriction fragment length polymorphism ( RFLP ) level . We have investigated the distribution , the length polymorphism , the informativeness , and the efficiency of analysis , of simple-sequence tandem repeats in the mouse cytokine genes . Highly polymorphic sequences have been identified in the P01584 , IL-1ra , P60568 , P05112 , P05113 , P05231 , P13232 , and P01579 genes . The utility and the value of these sequences as gene markers is exemplified by mapping the P13232 gene to mouse chromosome 3 close to pgk-1ps3 and Car-2 loci and the P01579 gene to chromosome 10 near the pg locus . Advantages of short tandemly repeated sequences as genetic markers are discussed in comparison with RFLPs . Neuroprotection mediated via neurotrophic factors and induction of neurotrophic factors . Neurotrophins and other neurotrophic factors have been shown to support the survival and differentiation of many neuronal populations of the central and peripheral nervous system . Therefore , administering neurotrophic factors could represent an alternative strategy for the treatment of acute and chronic brain disorders . However , the delivery of neurotrophic factors to the brain is one of the largest obstacles in the development of effective therapy for neurodegenerative disorders , because these proteins are not able to cross the blood-brain barrier . The induction of growth factor synthesis in the brain tissue by systemically administered lipophilic drugs , such as beta-adrenoceptor agonists , shown to increase endogenous nerve growth factor ( P01138 ) synthesis in the brain , would be an elegant way to overcome these problems of application . Stimulation of beta-adrenoceptors with clenbuterol led to increased P01138 synthesis in cultured central nervous system ( CNS ) cells and rat brain tissue . DB01407 -induced P01138 expression was reduced to the control levels by coadministration of beta-adrenoceptor antagonist propranolol . Furthermore , clenbuterol protected rat hippocampal neurons subjected to excitotoxic damage . The neuroprotective effect of clenbuterol in vitro depended on increased P01138 synthesis , since the neuroprotection was abolished by P01138 antisense oligonucleotide as well as by antibodies directed against P01138 itself . In vivo , clenbuterol protected rat hippocampus in a model of transient forebrain ischemia and reduced the infarct volume in a rat model of permanent middle cerebral artery occlusion ( MCAo ) . The neuroprotective effect of clenbuterol in vivo was accompanied by enhanced P01138 synthesis in brain tissue . These findings support our hypothesis that orally active P01138 inducers may have a potential as therapeutic agents for the treatment of neurodegenerative disorders and stroke . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Stimulation of central β2-adrenoceptors suppresses NFκB activity in rat brain : a role for IκB . In this study we examined the impact of systemic treatment with the long-acting brain penetrant β2-adrenoceptor agonist clenbuterol on NFκB activity and IκB expression in rat brain . DB01407 decreased NFκB activity ( p65 DNA binding ) in nuclear extracts prepared from rat cortex and hippocampus for up to 8h following a single treatment . This was accompanied by increased expression of IκBα mRNA and protein . The temporal increase in IκB protein expression paralleled the suppression of NFκB activity , suggesting that IκBα mediates the suppression NFκB activity observed . These actions of clenbuterol were prevented by pre-treatment with the non-selective β-adrenoceptor antagonist propranolol , the β2-adrenoceptor antagonist ICI-118,551 , but not the β1-adrenoceptor antagonist metoprolol , suggesting that the effects of clenbuterol on IκBα expression and NFκB activity are mediated specifically by the β2-adrenoceptor . In addition , the actions of clenbuterol were mimicked by systemic administration of another highly selective long-acting β2-adrenoceptor agonist formoterol . As neurodegenerative diseases are associated with inflammation we determined if clenbuterol could suppress NFκB activation that occurs in response to an inflammatory stimulus . In this regard we demonstrate that clenbuterol inhibited IκB phosphorylation and IκB degradation and inhibited NFκB activity in hippocampus and cortex of rats following a central injection of the inflammagen bacterial lipopolysaccharide ( LPS ) . In tandem , clenbuterol blocked expression of the NFκB-inducible genes P01375 -α and P05362 following LPS administration . Our finding that clenbuterol and formoterol inhibit NFκB activity in the CNS further supports the idea that β2-adrenoceptors may be an attractive target for treating neuroinflammation and combating inflammation-related neurodegeneration . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Erosive arthritis and hepatic granuloma formation induced by peptidoglycan polysaccharide in rats is aggravated by prasugrel treatment . Administration of the thienopyridine Q9H244 receptor antagonist , clopidogrel , increased the erosive arthritis induced by peptidoglycan polysaccharide ( PG-PS ) in rats or by injection of the arthritogenic K/BxN serum in mice . To determine if the detrimental effects are caused exclusively by clopidogrel , we evaluated prasugrel , a third-generation thienopyridine pro-drug , that contrary to clopidogrel is mostly metabolized into its active metabolite in the intestine . Prasugrel effects were examined on the PG-PS-induced arthritis rat model . Erosive arthritis was induced in Lewis rats followed by treatment with prasugrel for 21 days . Prasugrel treated arthritic animals showed a significant increase in the inflammatory response , compared with untreated arthritic rats , in terms of augmented macroscopic joint diameter associated with significant signs of inflammation , histomorphometric measurements of the hind joints and elevated platelet number . Moreover , fibrosis at the pannus , assessed by immunofluorescence of connective tissue growth factor , was increased in arthritic rats treated with prasugrel . In addition to the arthritic manifestations , hepatomegaly , liver granulomas and giant cell formation were observed after PG-PS induction and even more after prasugrel exposure . Cytokine plasma levels of P01584 , P05231 , MIP1 alpha , MCP1 , Q16552 and RANTES were increased in arthritis-induced animals . P22301 plasma levels were significantly decreased in animals treated with prasugrel . Overall , prasugrel enhances inflammation in joints and liver of this animal model . Since prasugrel metabolites inhibit neutrophil function ex-vivo and the effects of both clopidogrel and prasugrel metabolites on platelets are identical , we conclude that the thienopyridines metabolites might exert non-platelet effects on other immune cells to aggravate inflammation . Induction of nerve growth factor and basic fibroblast growth factor mRNA following clenbuterol : contrasting anatomical and cellular localization . RNase protection assay and in situ hybridization were used to analyze the temporal and cellular changes in nerve growth factor ( P01138 ) and basic fibroblast growth factor ( P09038 ) mRNA content evoked by the lipophilic beta-adrenergic receptor agonist clenbuterol in adult rat brain . DB01407 elicited a threefold increase in P01138 mRNA expression which was limited to the cerebral cortex . This increase was maximal at 5 h , still evident by 10 h , and declined to control levels by 24 h . By 10 h P01138 protein was also increased . Elevated P01138 mRNA hybridization following clenbuterol was localized in the superficial cortical layers II and III in large Nissl-pale cells , suggesting that P01138 mRNA induction occurs in neurons . In the same animals , clenbuterol induced a twofold increase in the levels of P09038 mRNA in cerebral cortex and hippocampus . This increase was localized primarily in glial cells as demonstrated by P09038 mRNA hybridization over all cortical regions and by labeling of the stratum lacunosum moleculare of the hippocampus . Our results suggest that enhanced noradrenergic tone regulates expression of these two trophic factors by different synaptic mechanisms and suggest that neurotransmitter(s) can coordinate trophic influences on different cell populations . Effect of isoproterenol and dexamethasone on the lipopolysaccharide induced expression of CD11b on bovine neutrophils . The present experiments investigate the changes in expression of CD11b on bovine neutrophils and its modulation by isopropylnoradrenaline ( IPN , isoproterenol ) , dexamethasone ( DX ) , phenylephrine ( alpha-agonist ) and clenbuterol ( beta-agonist ) . Both IPN and DX caused a dose-dependent inhibition of LPS-induced CD11b expression . A combination of IPN and DX elicited a synergistical decrease of the CD11b expression . DB01407 mimicked the effect of IPN , whereas phenylephrine did not . The effect of IPN and DX could at least partly be mediated through a decreased P01375 production by monocytes since tumor necrosis factor-alpha ( P01375 ) is shown to mediate a dose-dependent CD11b up-regulation . Stimulation of stress hormone receptors partly immuno-suppresses neutrophil functions by inhibition of CD11b expression on the neutrophil surface upon LPS stimulation . This inhibition is probably related to a decrease in P01375 production . A similar mechanism of immuno-suppression could contribute to the higher susceptibility of cattle to Gram-negative bacterial infections of the udder and lung during periods of stress . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . [ New pharmacological approaches to the treatment of schizophrenia ] . Schizophrenia is a serious mental disorder with a challenging rational pharmacotherapy . Neurochemical transmission in the dopaminergic system , especially via D2 receptors , and related changes in postsynaptic signal transduction are very important in both the formation of schizophrenia and current pharmacotherapeutic treatment with antipsychotic drugs . Blocking the serotonergic 5- Q13049 and P28335 receptors is growing growing importance with regard to the action mechanisms of new generation antipsychotic medications . Recent preclinical and clinical data show that dysfunction of central neurotrophins , such as nerve growth factor ( P01138 ) , brain-derived neurotrophic factor ( P23560 ) , and neurophin-3 ( P20783 ) might contribute to impaired brain development and neuroplasticity , leading to schizophrenia . In addition , some recent studies suggest that there is an important relationship between alcohol and substance addiction , and schizophrenia . There is also some preclinical data indicating that the central nitrergic system and agmatine(3/4)a biologically active agent produced after decarboxylation of arginine(3/4)might be interesting and important targets for understanding the etiopathogenesis of schizophrenia and for development of new drugs . Selective dopamine D3 receptor antagonists , specific agonists for metabotropic and DB01221 receptors of the glutamatergic system , and nicotinic alpha-7 receptor agonists were reported in preclinical and a limited number of clinical studies as potential new targets for schizophrenia treatment . In this review , new advances in the pharmacotherapy of schizophrenia and possible new targets are discussed in the light of the current literature . DB00588 -induced regulation of the balance within macrophage subpopulations . In asthma , treatment with inhaled corticosteroids reduces chronic peribronchial inflammation and restores the balance within macrophage subpopulations . This study investigates whether corticosteroids can regulate monocyte differentiation in vitro and thereby influence the balance of functionally distinct macrophages . Graded doses of fluticasone propionate ( FP ) were added to cultures of normal peripheral blood monocytes in the presence or absence of P05112 . Cells were harvested after 7 days ' culture . Double immunofluorescence studies were performed on cytospins of differentiated macrophages using the MoAbs RFD1 and RFD7 to distinguish inductive and suppressive macrophages by their respective phenotypes . Macrophage function was determined by quantifying allostimulation in a mixed leucocyte reaction and by measuring tumour necrosis factor-alpha ( P01375 ) production . FP reduced the number of mature cells with a D1+ antigen-presenting phenotype and up-regulated the development of cells with the D1/D7+ and D7+ phenotypes . Functionally , this was associated with reduced stimulation of T cell proliferation in a mixed leucocyte reaction ( P08235 ) . DB00588 also reversed the increase in both D1+ expression and P01375 production induced by P05112 . The effect of FP persisted for 24 h after removal of FP from the culture medium . These results suggest that FP treatment of asthmatics may have a direct beneficial effect by normalizing the macrophage subset imbalance that contributes to the chronic peribronchial inflammation present in this condition . Pharmacological modulation of nerve growth factor synthesis : a mechanistic comparison of vitamin D receptor and beta(2)-adrenoceptor agonists . Increasing nerve growth factor ( P01138 ) in the PNS is a rational strategy for treating certain neurodegenerative disorders . The present studies were undertaken to compare two compounds , a vitamin D(3) analogue ( CB1093 ) with minimal calcaemic effects , and clenbuterol , a long-acting beta(2)-adrenoceptor agonist , both of which induce P01138 synthesis in vivo . DB01407 caused significant increases in both P01138 mRNA and protein in 3T3 cells ; with maxima at 10 nM and at 8-12 h exposure . Effects of clenbuterol on P01138 mRNA were antagonized by propranolol . Mobility shift assays on whole cell extracts showed that clenbuterol increased P05412 binding in 3T3 cells prior to increasing P01138 synthesis . DB01407 was without effect on P01138 mRNA levels in L929 cells , whereas CB1093 caused significant increases in both P01138 mRNA and protein levels in both 3T3 and L929 cells . Stimulation was almost maximal at 24 h exposure and was sustained for at least 72 h . The magnitude of the increase was much greater in L929 ( 700 % increase ) than in 3T3 cells ( 80 % ) . Binding to the vitamin D nuclear receptor ( P11473 ) , which acts as a transcription factor itself , was increased as early as 30 min after exposure to of CB1093 and maintained up to 24 h . Increased P11473 binding preceded increased P01138 mRNA . A 150 % increase in AP-1 binding was also evident . This study demonstrates that CB1093 and clenbuterol stimulate P01138 levels in vitro and that AP-1 binding could be a commonality between the mechanism of P01138 induction of these two compounds . Nucleotide receptors stimulation by extracellular DB00171 controls Hsp90 expression through P27695 /Ref-1 in thyroid cancer cells : a novel tumorigenic pathway . Nucleotide receptors signaling affects cell proliferation , with possible implications on tumorigenic processes . However , molecular targets and action mechanisms of the extracellular nucleotides are still poorly elucidated . We have previously shown in ARO cells that P27695 /Ref-1 , a transcriptional coactivator responsible for the maintenance of the cellular proliferative rate , is functionally controlled by P2-mediated signaling . Here , we demonstrate that extracellular DB00171 has a mitogenic effect on ARO cells , increasing P29323 phosphorylation , P05412 activation , and cyclin D1 expression . Using the DB00171 /ADPase apyrase and the P2 receptor antagonist suramin , we show that the extracellular DB00171 , physiologically released by ARO cells , exerts mitogenic effects . A differential proteomic approach was used to identify molecular events associated with the DB00171 -induced cell proliferation . Among other proteins , Hsp90 was found upregulated upon DB00171 stimulation . Pretreatment with suramin completely blocked the DB00171 -induced Hsp90 activation , confirming the involvement of cell-surface P2 nucleotide receptors in the DB00171 -mediated activation of ARO cells . Treatment of proliferating ARO cells with suramin and apyrase significantly reduced the intracellular levels of Hsp90 , suggesting an autocrine/paracrine mechanism of control on Hsp90 expression by extracellular DB00171 . The influence of Hsp90 on DB00171 -induced cell proliferation was also demonstrated by its specific inhibition with 17- P29372 . The molecular pathway by which DB00171 stimulates cell proliferation was further investigated by siRNA strategies showing that Hsp90 is a target of P27695 /Ref-1 functional activation . Stimulation of ARO cells with specific nucleotide receptors agonists evidenced a major involvement of P47900 and P41231 receptors in controlling the Hsp90 activation . Accordingly , these two receptors resulted significantly upregulated in sample biopsies from different thyroid tumors . Regulatory roles of sex hormones in cutaneous biology and immunology . Recent studies have revealed that sex hormones manifest a variety of biological and immunological effects in the skin . Pregnancy , menstruation and the menopause modulate the natural course of psoriasis , indicating a female hormone-induced regulation of skin inflammation . Estrogen in vitro down-regulates the production of the neutrophil , type 1 T cell and macrophage-attracting chemokines , P10145 , P02778 , P13501 , by keratinocytes , and suppresses IL-12 production and antigen-presenting capacity while enhancing anti-inflammatory P22301 production by dendritic cells . These data indicate that estrogen may attenuate inflammation in psoriatic lesions . Estrogen , alone or together with progesterone , prevents or reverses skin atrophy , dryness and wrinkles associated with chronological or photo-aging . Estrogen and progesterone stimulate proliferation of keratinocytes while estrogen suppresses apoptosis and thus prevents epidermal atrophy . Estrogen also enhances collagen synthesis , and estrogen and progesterone suppress collagenolysis by reducing matrix metalloproteinase activity in fibroblasts , thereby maintaining skin thickness . Estrogen maintains skin moisture by increasing acid mucopolysaccharide or hyaluronic acid levels in the dermis . Progesterone increases sebum secretion . Estrogen accelerates cutaneous wound healing stimulating P01138 production in macrophages , GM- P04141 production in keratinocytes and P09038 and TGF-beta1 production in fibroblasts , leading to the enhancement of wound re-innervation , re-epithelialization and granulation tissue formation . In contrast , androgens prolong inflammation , reduce deposition of extracellular matrix in wounds , and reduce the rate of wound healing . Estrogen enhances P15692 production in macrophages , an effect that is antagonized by androgens and which may be related to the development of granuloma pyogenicum during pregnancy . These regulatory effects of sex steroids may be manipulated as therapeutic or prophylactic measures in psoriasis , aging , chronic wounds or granuloma pyogenicum . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . Cell cycle arrest mediated by the MEK/mitogen-activated protein kinase pathway . The mitogen-activated protein kinase ( MAPK ) cascade plays a crucial role in the transduction of extracellular signals into responses governing growth and differentiation . The effects of a specific inhibitor of the MAPK kinase ( MEK ) /MAPK pathway ( PD98059 ) on nerve growth factor ( P01138 ) -induced growth arrest and inhibition of cell cycle-dependent kinases ( CDKs ) have been examined . Treatment of NIH 3T3 cells expressing P04629 with PD98059 dramatically reversed the complete inhibition of growth of these cells caused by P01138 . PD98059 also blocked the ability of P01138 to inhibit the activities of P11802 and P24941 , while partially preventing P01138 induction of p21Cip1/ P38936 . To independently evaluate the involvement of the MEK/MAPK pathway in growth arrest , an inducible activated form of the P04049 protooncogene ( delta RAF-1:ER ) was expressed in these cells . Activation of delta RAF-1:ER resulted in a prolonged increase in MAPK activity and growth arrest of these cells , with concomitant induction of p21Cip1/ P38936 and inhibition of P24941 activity . These effects of delta RAF-1:ER activation were all reversed by treatment of cells with PD98059 . These data indicate that in addition to functioning as a positive effector of growth , stimulation of the MEK/MAPK pathway can result in an inhibition of CDK activity and cell cycle arrest . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . The β2-adrenoceptor agonist clenbuterol elicits neuroprotective , anti-inflammatory and neurotrophic actions in the kainic acid model of excitotoxicity . Excitotoxicity is a mechanism of neuronal cell death implicated in a range of neurodegenerative conditions . Systemic administration of the excitotoxin kainic acid ( KA ) induces inflammation and apoptosis in the hippocampus , resulting in neuronal loss . Evidence indicates that stimulation of glial β(2)-adrenoceptors has anti-inflammatory and neurotrophic properties that could result in neuroprotection . Consequently , in this study we examined the effect of the β(2)-adrenoceptor agonist clenbuterol on KA-induced inflammation , neurotrophic factor expression and apoptosis in the hippocampus . DB01407 ( 0.5mg/kg ) was administered to rats one hour prior to KA ( 10mg/kg ) . Epileptic behaviour induced by KA was assessed for three hours following administration using the Racine scale . Twenty-four hours later TUNEL staining in the P07451 hippocampal subfield and hippocampal caspase-3 activity was assessed to measure KA-induced apoptosis . In addition , expression of inflammatory cytokines ( IL-1β and IFN-γ ) , inducible nitric oxide synthase ( P35228 ) , kynurenine pathway enzymes indolamine 2,3-dioxygenase ( P14902 ) and kynurenine monooxygenase ( O15229 ) , the microglial activation marker CD11b , and the neurotrophins P23560 and P01138 were quantified in the hippocampus using real-time PCR . Whilst clenbuterol treatment did not significantly alter KA-induced epileptic behavior it ameliorated KA-induced apoptosis , and this neuroprotective effect was accompanied by reduced inflammatory cytokine expression , reduced expression of P35228 , P14902 , O15229 and CD11b , coupled with increased P23560 and P01138 expression in KA-treated rats . In conclusion , the β(2)-adrenoceptor agonist clenbuterol has anti-inflammatory and neurotrophic actions and elicits a neuroprotective effect in the KA model of neurodegeneration . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . Ceramide is responsible for the failure of compensatory nerve sprouting in apolipoprotein E knock-out mice . P02649 ( apoE ) is a key transporter of the cholesterol and phospholipids required for membrane synthesis and nerve growth . We now report a virtual absence in apoE knock-out ( KO ) mice of normal nerve growth factor ( P01138 ) -driven compensatory sprouting of undamaged cutaneous nociceptive nerves . In contrast , P01138 -independent regeneration of crushed axons was unaffected . Essentially similar results came from aged wild-type mice . In apoE KO mice , the endogenous sprouting stimulus was suspect , because P01138 administration induced normal sprouting ; nevertheless , P01138 increased normally in denervated skin , transported normally in the axons , and led to phosphorylation of trkA , erk1 , and erk2 . However , sprouting was restored in apoE KO mice ( although not in aged mice ) by fumonisin B1 , an inhibitor of ceramide synthesis . A shotgun analysis revealed a wide array of changes in individual ceramide species in Q86YR7 neurons of apoE KO mice , and the changes for ceramide species OH_N15:0 made it a candidate inhibitor of sprouting ( increased in apoE KO mice and normalized by fumonisin B1 ) . Nevertheless , the unknown effects of individual ceramide species on sprouting , as well as the variability of their changed levels in apoE KO mice and how these were affected by fumonisin B1 , support a different conclusion . We suggest that absence of apoE expression alters the balance among ceramide species to one that collectively inhibits compensatory sprouting , whereas fumonisin B1 establishes a new balance that allows sprouting . Nontoxic ceramide modulators might usefully promote sprouting and circuitry repair in neurodegenerative disorders in which ceramide species are perturbed , adding to the benefits of reducing ceramide-induced neuronal apoptosis . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Monoclonal antibodies targeting P01584 reduce biomarkers of atherosclerosis in vitro and inhibit atherosclerotic plaque formation in P02649 -deficient mice . OBJECTIVE : Atherosclerosis is a condition that is increasingly contributing to worldwide mortality through complications such as stroke and myocardial infarction . IL-1β plays multiple direct , local roles in the formation and stability of the atheroma by eliciting the production of additional cytokines and proteolytic enzymes from macrophages , endothelial cells ( EC ) and smooth muscle cells ( SMC ) . We therefore tested whether an anti-IL-1β antibody , DB06062 , might inhibit the secretion of pro-atherogenic cytokines from macrophages in vitro and affect a positive outcome in the P02649 -deficient mouse ( ApoE(-/-) ) model of atherosclerosis in vivo . METHODS AND RESULTS : In an in vitro co-culture model , DB06062 inhibited macrophage-induced secretion of key atherogenic cytokines from EC and SMC , including P05231 , P10145 , P13500 and TNFα . The release of degradative enzymes , such as the matrix metalloproteinases P08254 and P14780 , was also decreased by DB06062 . In addition , DB06062 inhibited the secretion of P13232 from EC and P05112 from SMC , cytokines not previously reported to be driven by IL-1β in this context . In vivo , XMA052 MG1K , a chimeric murine version of DB06062 , inhibited the formation of atherosclerotic lesions in the ApoE(-/-) model at all three doses tested . This effect was comparable to that reported for complete genetic ablation of IL-1β or IL-1R1 on an ApoE(-/-) background and was associated with decreases in plasma non-HDL/HDL cholesterol ratio and plaque lipid content and macrophage infiltration . CONCLUSIONS : These results demonstrate for the first time that an antibody targeting IL-1β can inhibit the progression of atherosclerosis in vivo , highlighting the importance of this key cytokine in cardiovascular disease . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . Expression of cytokine/growth factors and their receptors in human melanoma and melanocytes . Human tumors can constitutively express cytokines and growth factors , but the extent of this expression has not been investigated . Using 44 different probes to cytokines , growth factors , and their receptors , we tested 21 melanoma and 5 melanocyte cultures for RNA transcript expression by reverse transcriptase-polymerase chain reaction . With 30 amplification cycles , expression of the cytokines interleukin ( IL ) -1 beta , P05231 , leukemia inhibitory factor ( P15018 ) , P13232 , gro alpha , P10145 and the p35 chain of IL-12 was detected in more than 60 % of melanomas . Concomitant receptors for P05231 and P13232 were also detected . P01583 , P05113 , Rantes , P22301 , interferon ( IFN ) -beta , tumor-necrosis factor ( P01375 ) -alpha , G-colony-stimulating factor ( P04141 ) and GM- P04141 were expressed at lower levels . Melanocytes showed greatly reduced cytokine RNA transcripts , and only gro alpha was consistently detected . No expression of P60568 , P08700 , P05112 , P15248 , the p40 chain of IL-12 , IFN-alpha or P01579 RNA transcripts was detected in melanomas or melanocytes . The growth factors expressed by melanomas and , after further signal amplification , by melanocytes were transforming growth factor ( TGF ) -alpha , epidermal growth factor ( P01133 ) , TGF-beta , endothelial-cell growth factor ( P05230 ) , basic-fibroblast growth factor ( P09038 ) , nerve growth factor ( P01138 ) and steel . The receptors P00533 , FGFR , NGFRp70 and c-kit were also expressed by melanomas and melanocytes . These results point to new possible autocrine and paracrine pathways in melanoma biology . Combined anti-inflammatory effects of β2-adrenergic agonists and DB05876 inhibitors on astrocytes by upregulation of intracellular DB02527 . Inflammation is an important hallmark of all neurodegenerative diseases and activation of different glial populations may be involved in the progression of some of these disorders . Especially , the activation of astroglia can lead to long-term detrimental morphological changes , such as scar formation . Therefore , improved strategies to modulate inflammation in these cells are currently being investigated . We investigated the interaction of phosphodiesterase ( PDE ) 4 inhibitors , such as rolipram , with other agents raising cellular DB02527 levels . When used alone , none of the DB05876 inhibitors increased DB02527 levels . The adenylate cyclase activator forskolin , the β(2)-adrenergic agonist clenbuterol and the mixed β(1)/β(2)-adrenergic agonist isoproterenol increased intracellular DB02527 levels of cortical murine astrocytes . This increase was synergistically elevated by rolipram or the DB05876 inhibitor RO-201724 , but not by inhibition of PDE3 . Inflammatory stimulation of the cells with the cytokines P01375 -α , IL-1β and IFN-γ strongly induced Q07343 and augmented overall DB05876 activity , while PDE3 activity was low . DB01407 and forskolin caused downregulation of cytokines and chemokines such as P05231 and P13500 . This effect was further enhanced by rolipram , but not by the PDE3 inhibitor milrinone . The DB02527 -raising drug combinations attenuated the upregulation of P01375 -α and P05231 mRNA and the secretion of P05231 , but did not affect initial NF-κB signalling triggered by the stimulating cytokines . These results indicate that DB05876 may be a valuable anti-inflammatory target in brain diseases , especially under conditions associated with stimulation of DB02527 -augmenting astrocyte receptors as is observed by clenbuterol treatment . Apotransferrin decreases the response of oligodendrocyte progenitors to PDGF and inhibits the progression of the cell cycle . In the CNS , transferrin ( Tf ) is expressed by the oligodendroglial cells ( OLGcs ) and is essential for their development . We have previously shown that apotransferrin ( aTf ) accelerates maturation of OLGcs in vivo as well as in vitro . The mechanisms involved in this action appear to be complex and have not been completely elucidated . The aim of this study was to investigate if Tf participates in the regulation of the cell cycle of oligodendroglial progenitor cells ( OPcs ) . Primary cultures of OPcs were treated with aTf and/or with different combinations of mitogenic factors . Cell cycle progression was studied by BrdU incorporation , flow cytometry and by the expression of cell cycle regulatory proteins . Apotransferrin decreased the number of BrdU+ cells , increasing the cell cycle time and decreasing the number of cells in S phase . The cell cycle inhibitors p27kip1 , p21cip1 and p53 were increased , and in agreement with these results , the activity of the complexes involved in P55008 -S progression ( cyclin D/ P11802 , cyclin E/ P24941 ) , was dramatically decreased . Apotransferrin also inhibited the mitogenic effects of PDGF and PDGF/IGF on OPcs , but did not affect their proliferation rate in the presence of P09038 , P09038 /PDGF or P09038 /IGF . Our results indicate that inhibition of the progression of the cell cycle of OPcs by aTf , even in the presence of PDGF , leads to an early beginning of the differentiation program , evaluated by different maturation markers ( O4 , GC and MBP ) and by morphological criteria . The modulation by aTf of the response of OPcs to PDGF supports the idea that this glycoprotein might act as a key regulator of the OLGc lineage progression . Transcriptional profiles during the differentiation and maturation of monocyte-derived dendritic cells , analyzed using focused microarrays . Dendritic cells ( DC ) are professional antigen-presenting cells capable of initiating primary immune responses . They have been intensively studied and are used in both basic immunology research and clinical immunotherapy . However , the genetic pathways leading to DC differentiation and maturation remain poorly understood . Using focused microarrays with oligonucletotide probes for 120 genes encoding co-stimulatory molecules , chemokines , chemokine receptors , cytokines , cytokine receptors , TLRs , and several other related molecules , we analyzed the kinetics of gene expression for the overall differentiation process of monocytes into mature DC . In parallel , we compared the transcriptional profiles in DC maturation in the presence of LPS , P01375 or trimeric P29965 . We found similar transcriptional profiles for early immature DC and immature DC , respectively generated by culturing monocytes with GM- P04141 and P05112 for three or six days . We identified sets of common and stimuli-specific genes , the expression of which changed following stimulation with LPS , P01375 or P29965 . A dynamic analysis of the entire DC differentiation and maturation process showed that some important inflammatory and constitutive chemokines are transcribed in both immature and mature DC . The correlative expression kinetics of the gene pairs P14778 / P27930 , P40933 / Q13261 , Q9NNX6 / P13598 and Q9NNX6 / P32942 imply that they all play crucial roles in mediating DC functions . Thus , our analysis with focused microarrays shed light on the transcriptional kinetics of DC differentiation and maturation , and this method may also prove useful for identifying novel marker genes involved in DC functions . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . Evaluation of human astrocytoma and glioblastoma cell lines for nerve growth factor release . Nerve growth factor ( P01138 ) prevents degeneration of cholinergic neurons in the central nervous system ( CNS ) , and has potential as a therapeutic treatment for Alzheimer 's disease . The inability of P01138 to cross the blood-brain barrier has prompted pharmacological approaches investigating peripherally administered compounds that stimulate release of endogenous P01138 . This study describes the P01138 -releasing properties of six human astrocytoma and glioblastoma cell lines ( SW 1088 , SW 1783 and CRL 1718 astrocytomas , and U-138 , U-373 , and T98G glioblastomas ) . Using a highly specific two-site ELISA for human P01138 , basal P01138 release could be detected in all cell lines , with the lowest level in the T98G line ( approximately 80 pg P01138 /ml ) . Cell lines tested with a variety of compounds for 24 h in serum-free media demonstrated stimulation of P01138 release by distinct mechanisms . P01138 levels were markedly elevated ( up to 8-fold above vehicle-treated cells ) when stimulated with the cytokine interleukin-1 beta ( P01584 ) . Phorbol ester stimulated P01138 release 4-fold . DB01407 , 4-methyl catechol , and propentofylline had little activity , while 6-(4-hydroxybutyl)-2,3,5,-trimethyl-1,4,benzoquinone ( DB01157 ) , dexamethasone and 1,25-dihydroxyvitamin D3 elevated P01138 levels 3-fold . These data indicate differences in the ability of human astrocytoma and glioblastoma cells to release P01138 when stimulated with mechanistically distinct compounds . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . P40933 affects serotonin system and exerts antidepressive effects through IL15Rα receptor . Contrary to the reduction of depressive-like behavior observed in several strains of cytokine receptor knockout mice , mice lacking the specific receptor for interleukin ( IL ) -15 showed increased immobility in tail suspension and modified forced swimming tests . There was also a reduction in social interactions . The hippocampus of the IL15Rα knockout mice had decreased mRNA for 5-HT(1A) , increased mRNA for 5-HT(2C) , and region-specific changes of serotonin reuptake transporter ( P31645 ) immunoreactivity . DB00472 ( the classic antidepressant DB00472 , which inhibits 5-HT(2C) and P31645 ) reduced the immobility of the IL15Rα knockout mice in comparison with their pretreatment baseline . Together with the unchanged performance of the IL15Rα knockout mice on the rotarod , this response to fluoxetine indicates that the immobility reflects depression . Wildtype mice responded to P40933 treatment with improvement of immobility induced by forced swimming , whereas the knockout mice failed to respond . Thus , the cognate P40933 receptor is necessary for the antidepressive activity of P40933 . In ex vivo studies , P40933 decreased synaptosomal uptake of 5-HT , and modulated the expression of 5-HT(2C) and P31645 in cultured neurons in a dose- and time-dependent manner . Thus , the effect of P40933 on serotonin transmission may underlie the depressive-like behavior of IL15Rα knockout mice . We speculate that P40933 is essential to maintain neurochemical homeostasis and thereby plays a role in preventing neuropsychiatric symptoms . Regulation of P14061 and P18405 in lymphocytes . We previously reported lymphocyte expression of genes encoding enzymes required for steroid metabolism ; however , only 17beta-HSD and 5alpha-reductase showed significant enzyme activity . We now investigate regulation of lymphocyte expression for genes encoding 17beta-HSD and 5alpha-reductase . Cultured human T and B lymphoid cell lines and peripheral blood mononuclear cells were treated with known regulators of steroidogenic gene expression including forskolin , PMA , ionomycin , various steroids , interleukin ( IL ) -4 , and P05231 . Treatment with 10 or 50 microM forskolin resulted in a 20-60 % reduction of expression for P14061 ( encoding 17beta-HSD I ) in T and B lymphoid cell lines and peripheral blood mononuclear cells , although such a change was not observed in the expression of P18405 ( encoding 5alpha-reductase I ) . No significant changes were found when cells were treated for 24 h with various concentrations of PMA or ionomycin . Incubation with 10(-9) to 10(-7) M androstenedione or estradiol increased expression of P14061 , while testosterone decreased the expression of this gene . P18405 expression was increased in the presence of 5alpha- DB02901 although no consistent changes were observed when the cells were treated with testosterone . Other steroids , including dexamethasone , progesterone , and 6-hydroxypregnanolone , produced no effects on expression of either P14061 or P18405 . Treatment with 0.1-10 ng/ml of P05112 or P05231 also did not effect significant changes in gene expression . These data implicate the involvement of the DB02527 -protein kinase signal transduction pathway in regulating lymphocyte expression of P14061 . Furthermore , it appears that lymphocyte P14061 and P18405 are regulated to some extent by specific steroids . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells .

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interacts_with DB01088?

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8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . P43119 -induced P40763 phosphorylation in human erythroleukemia cells is mediated via Galpha(s) and Galpha(16) hybrid signaling . Human prostacyclin receptor ( hIP ) stimulates P40763 via pertussis toxin-insensitive G proteins in human erythroleukemia ( HEL ) cells . Since hIP can utilize G(s) and G(q) proteins for signal transduction and that both G proteins can induce P40763 phosphorylation and activation via complex signaling networks , we sought to determine if one of them is predominant in mediating the hIP signal . Stimulation of P40763 DB00135 (705) and DB00133 (727) phosphorylations by the IP-specific agonist , cicaprost , was sensitive to inhibition of protein kinase A , phospholipase Cbeta , protein kinase C , calmodulin-dependent protein kinase II and O60674 /3 . Unlike Galpha(16)-mediated regulation of P40763 in the same cells , cicaprost-induced P40763 DB00135 (705) phosphorylation was resistant to inhibition of Src and MEK while P40763 DB00133 (727) phosphorylation distinctly required phosphatidylinositol-3 kinase . This unique inhibitor-sensitivity pattern of P40763 phosphorylation was reproduced in HEL cells by stimulating the G(16)-coupled C5a receptor in the presence of dibutyryl- DB02527 , suggesting that the change in inhibitor-sensitivity was due to activation of the G(s) pathway . This postulation was confirmed by expressing constitutively active Galpha(16)QL and Galpha(s)QL in human embryonic kidney 293 cells and the inhibitor-sensitivity of Galpha(16)QL-induced P40763 phosphorylations could be converted by the mere presence of Galpha(s)QL to resemble that obtained with cicaprost in HEL cells . In addition , the restoration of the Galpha(16)-mediated inhibitor-sensitivity upon cicaprost induction in Galpha(s)-knocked down HEL cells again verified the pivotal role of G(s) signal . Taken together , our observations illustrate that co-stimulation of G(s) and G(q) can result in the fine-tuning of P40763 activation status , and this may provide the basis for cell type-specific responses following activation of hIP . DB05229 sodium , a stable prostacyclin analogue , elicits dilation of isolated porcine retinal arterioles : roles of P29474 and potassium channels . PURPOSE : DB01240 ( DB01240 ) is usually described as an endoEDRFsthelium-derived relaxing factor , but the vasoreactivity to DB01240 in the retinal arterioles and the underlying mechanisms are not fully understood . We examined the effects of DB01240 on the retinal microcirculation using beraprost sodium ( BPS ) , a stable DB01240 analogue , and the signaling mechanisms involved in this vasomotor activity . METHODS : Porcine retinal arterioles were isolated , cannulated , and pressurized without flow in vitro . Video microscopic techniques recorded the diametric responses to BPS . RESULTS : DB05229 sodium elicited dose-dependent ( 0.1 pM-0.1 μM ) vasodilation of the retinal arterioles that was abolished by the P43119 ( IP ) antagonist CAY10441 . DB05229 sodium-induced vasodilation decreased by 50 % after the endothelium was removed and was inhibited by the nitric oxide ( NO ) synthase inhibitor N(G)-nitro-L-arginine methyl ester ( L-NAME ) comparable with denudation . Inhibition of soluble guanylyl cyclase by 1H-1,2,4-oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) and blockage of protein kinase A ( PKA ) by Rp-8-Br-cAMPS were comparable to L-NAME . DB05229 sodium-induced vasodilation was also inhibited by the nonselective potassium channel inhibitor , tetraethylammonium , and the adenosine triphosphate-sensitive potassium ( KATP ) channel blocker , glibenclamide . Residual vasodilation in the presence of glibenclamide decreased further with subsequent application of ODQ . CONCLUSIONS : DB05229 sodium , a stable DB01240 analogue , causes vasodilation of the retinal arterioles mediated via the IP receptor . The current findings suggest that BPS elicits endothelium-dependent and -independent dilation of the retinal arterioles mediated by NO induced by activation of PKA in the endothelium and the KATP channel activation in the vascular smooth muscle , respectively . Two types of prostacyclin receptor coupling to stimulation of adenylate cyclase and phosphatidylinositol hydrolysis in a cultured mast cell line , BNu-2cl3 cells . DB01240 ( DB01240 ) -mediated signal transduction was examined in interleukin 3 ( P08700 ) -dependent BNu-2cl3 mast cells . DB01088 , a stable DB01240 analogue , induced the accumulation of intracellular DB02527 and IP3 , and an increase in the intracellular Ca2+ concentration . Pretreatment of the cells with a protein kinase C activator , 12-O-tetradecanoyl phorbol 13-acetate , suppressed the iloprost-induced IP3 accumulation and Ca2+ mobilization , but inversely potentiated the DB02527 accumulation , suggesting that neither of these signal transduction pathways of iloprosts is the result of a secondary effect of activation of the other . Removal of P08700 from the culture medium reduced the iloprost-induced IP3 accumulation and Ca2+ mobilization , while it had no effect on the iloprost-induced DB02527 accumulation at all . These results taken together suggest that BNu-2cl3 cells express two types of P43119 ; one couples to stimulation of adenylate cyclase , its expression being independent of P08700 , while the other couples to phosphatidylinositol hydrolysis , its expression being dependent on P08700 . Induction of prostacyclin receptor expression in human erythroleukemia cells . We have identified both high-affinity ( KD = 36 +/- 3 nM ) and low-affinity ( KD = 2.1 +/- 0.8 microM ) prostacyclin ( DB01240 ) -receptor sites on human erythroleukemia ( HEL ) cells using the radiolabelled prostacyclin analogue . [3H]iloprost . The addition of the phorbol ester , TPA , to the culture medium caused a 5-10-fold increase in the number of both the low- and the high-affinity sites , without any change in their affinity constants . DB01088 stimulated HEL cell membrane adenylate cyclase activity 5-fold . This stimulation was potentiated in the presence of GTP , indicating a conventional P43119 -G2-adenylate cyclase system . HEL cells represent a source of prostacyclin receptor mRNA which may be of value in expression cloning of this receptor . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . S-sulfhydration of Q02750 leads to P09874 activation and DNA damage repair . The repair of DNA damage is fundamental to normal cell development and replication . Hydrogen sulfide ( H2S ) is a novel gasotransmitter that has been reported to protect cellular aging . Here , we show that H2S attenuates DNA damage in human endothelial cells and fibroblasts by S-sulfhydrating Q02750 at cysteine 341 , which leads to P09874 activation . H2S-induced Q02750 S-sulfhydration facilitates the translocation of phosphorylated P27361 /2 into nucleus , where it activates P09874 through direct interaction . Mutation of Q02750 cysteine 341 inhibits P29323 phosphorylation and P09874 activation . In the presence of H2S , activated P09874 recruits P18887 and P49916 to DNA breaks to mediate DNA damage repair , and cells are protected from senescence . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . [ Thrombocyte prostacyclin receptors in gestational hypertension and pre-eclampsia ] . OBJECTIVE : To determine prostacyclin ( DB01240 ) receptor characteristics in pregnancies complicated by hypertension and to assess any relation to the clinical outcome . METHODS : Radioligand binding studies with [ 3H ] - DB01088 were performed to measure receptor capacity ( Bmax ) and affinity ( Kd-1 ) using platelet membranes from patients with preeclampsia , gestational hypertension or normal pregnancy . RESULTS : P43119 capacity did not differ between the patient groups . In contrast , P43119 affinity was diminished in gestational hypertension and considerably reduced in preeclampsia compared to normal pregnancy . A similar pattern was found in fetal growth ( normal pregnancy > gestational hypertension > preeclampsia ) . Furthermore , the rate of low Apgar scores and acidosis was increased in preeclampsia . CONCLUSIONS : In preeclampsia reduced platelet P43119 affinity was found as well as poor pregnancy outcome in comparison with normal pregnancy , whereas these differences were less pronounced in gestational hypertension . This suggests a role of DB01240 and its receptor in gestational hypertension and in particular in preeclampsia . Developmental regulation of prostacyclin synthase and prostacyclin receptors in the ovine uterus and conceptus during the peri-implantation period . This study documents the expression of prostacyclin ( DB01240 ) synthase ( Q16647 ) and DB01240 receptors in the trophoblast and uterus of the ewe at the time of maternal recognition of pregnancy ( i.e. days 7 , 9 , 12 , 14 and 17 ) . The membrane receptor for DB01240 ( P43119 ) and the nuclear receptors , i.e. peroxisome proliferator-activated receptors ( Q07869 ) and their heterodimer partners the retinoid X receptors ( RXR ) , were analysed . In the endometrium , Q16647 transcript and protein were expressed at day 9 of pregnancy and levels declined from days 12 to 17 . Immunohistochemistry and in situ hybridization indicated that Q16647 was mainly located in the luminal epithelium of the endometrium . Endometrial P43119 , Q07869 , P37231 and P48443 expression was regulated during the peri-implantation period whereas Q03181 , P19793 and P28702 were consistently expressed . In the trophoblast , Q16647 transcript levels rose as development progressed and peaked at day 17 . P43119 and Q07869 transcripts peaked before day 12 and then declined and became nearly undetectable by day 17 , whereas Q03181 and P37231 transcript levels rose steadily from days 12 to 17 . Because the PPARs and the RXRs display different expression profiles , we suggest that different heterodimers may form and support distinct functions as development proceeds . Our results also underline the importance of Q16647 and Q03181 in the trophoblast and P43119 in the uterus , suggesting that DB01240 is of both uterine and trophoblastic origin and is involved in a complex signalling pathway at around the time of implantation in the ewe . Expression of cytokine/growth factors and their receptors in human melanoma and melanocytes . Human tumors can constitutively express cytokines and growth factors , but the extent of this expression has not been investigated . Using 44 different probes to cytokines , growth factors , and their receptors , we tested 21 melanoma and 5 melanocyte cultures for RNA transcript expression by reverse transcriptase-polymerase chain reaction . With 30 amplification cycles , expression of the cytokines interleukin ( IL ) -1 beta , P05231 , leukemia inhibitory factor ( P15018 ) , P13232 , gro alpha , P10145 and the p35 chain of IL-12 was detected in more than 60 % of melanomas . Concomitant receptors for P05231 and P13232 were also detected . P01583 , P05113 , Rantes , P22301 , interferon ( IFN ) -beta , tumor-necrosis factor ( P01375 ) -alpha , G-colony-stimulating factor ( P04141 ) and GM- P04141 were expressed at lower levels . Melanocytes showed greatly reduced cytokine RNA transcripts , and only gro alpha was consistently detected . No expression of P60568 , P08700 , P05112 , P15248 , the p40 chain of IL-12 , IFN-alpha or P01579 RNA transcripts was detected in melanomas or melanocytes . The growth factors expressed by melanomas and , after further signal amplification , by melanocytes were transforming growth factor ( TGF ) -alpha , epidermal growth factor ( P01133 ) , TGF-beta , endothelial-cell growth factor ( P05230 ) , basic-fibroblast growth factor ( P09038 ) , nerve growth factor ( P01138 ) and steel . The receptors P00533 , FGFR , NGFRp70 and c-kit were also expressed by melanomas and melanocytes . These results point to new possible autocrine and paracrine pathways in melanoma biology . P43119 induces P42224 and P40763 phosphorylations in human erythroleukemia cells : a mechanism requiring PTX-insensitive G proteins , P29323 and JNK . The ability of the human prostacyclin receptor ( hIP ) to regulate the activities of signal transducers and activators of transcription ( STATs ) has not yet been documented . In the present study , we have delineated the mechanism by which hIP induces P40763 phosphorylations in human erythroleukemia ( HEL ) cells . Stimulation of endogenous hIP by its specific agonist , cicaprost , resulted in P40763 Tyr705 and Ser727 phosphorylations in a time- and concentration-dependent manner . Cicaprost-induced P40763 Tyr705 and Ser727 phosphorylations were resistant to pertussis toxin ( PTX ) treatment , suggesting that these responses were mediated through PTX-insensitive G proteins . In addition , extracellular signal-regulated kinase ( P29323 ) and c-Jun N-terminal kinase ( JNK ) , but not p38 MAPK , were shown to be phosphorylated by cicaprost in a time- and concentration-dependent manner via PTX-insensitive G proteins . The levels of the interaction between P40763 , P29323 and JNK were enhanced by cicaprost treatment . The involvement of P04049 , Q02750 /2 and JNK in cicaprost-induced phosphorylations of P40763 was illustrated by the use of their selective inhibitors . In contrast , p38 MAPK did not appear to be required . Similar observations were obtained with P42224 upon stimulation by cicaprost . Taken together , these results demonstrate for the first time that hIP activation by cicaprost can lead to P42224 and P40763 phosphorylations via signaling pathways involving PTX-insensitive G proteins , P29323 and JNK . The regulation of rotenone-induced inflammatory factor production by DB00171 -sensitive potassium channel expressed in BV-2 cells . Our previous studies have demonstrated that activating DB00171 -sensitive potassium channel ( K( DB00171 ) channel ) , not only improved Parkinsonian behavior and neurochemical symptoms , but also reduced P35228 activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson 's disease ( PD ) . In this study , it was first shown that the subunits of K( DB00171 ) channels are expressed in BV-2 cells , and then it was investigated whether K( DB00171 ) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone . It was found that K( DB00171 ) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and Q09428 2A/2B . K( DB00171 ) channel openers ( KCOs ) including pinacidil , diazoxide and iptakalim ( Ipt ) exerted beneficial effects on rotenone-induced morphological alterations of BV-2 cells , decreased tumor necrosis factor alpha ( P01375 ) production and the expression and activity of inducible isoform of nitric oxide synthase ( P35228 ) . Either glibenclamide or 5-hydroxydecanoate acid ( a selective mitochondrial K( DB00171 ) channel blocker ) could abolish the effects of KCOs , suggesting that K( DB00171 ) channels , especially mitochondrial DB00171 -sensitive potassium channels ( mitoK( DB00171 ) channels ) , played a crucial role in preventing the activation of BV-2 cells , and subsequently the production of a variety of proinflammatory factors . Therefore , activation of K( DB00171 ) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders . Expression of adhesion molecules and c-kit on P28906 + hematopoietic progenitor cells : comparison of cytokine-mobilized blood stem cells with normal bone marrow and peripheral blood . We assessed the expression of the adhesion molecules leukocyte function antigen-1 ( LFA-1 , CD11a ) , intercellular adhesion molecule-1 ( P05362 , CD54 ) , homing-associated cell adhesion molecule ( H- P62158 , P16070 ) , and c-kit ( stem cell factor receptor ) on the P28906 + progenitor population from the leukapheresis products of 23 patients ( LP P28906 + ) . For blood stem cell collection granulocyte colony-stimulating factor ( DB00099 ) or interleukin-3/granulocyte-macrophage colony-stimulating factor ( P08700 /GM- P04141 ) was administered after cytotoxic chemotherapy . Furthermore , bone marrow- and blood-derived P28906 + progenitor cells from 6 normal volunteers ( BM and PB P28906 + ) were analyzed . LFA-1 expression was higher on PB P28906 + ( 88.2 +/- 2.5 % , mean +/- SEM ) than on BM P28906 + ( 75.3 +/- 4.3 % ) . Following cytokine administration , LFA-1 was expressed on only 59.7 +/- 3.7 % of LP P28906 + at a low fluorescence intensity , suggesting that down-regulation of LFA-1 may facilitate the egress of cells from the bone marrow and prolong their circulation . In contrast , P05362 was weakly positive on P28906 + cells from all sources . P16070 was expressed on the vast majority of P28906 + cells ( > 95 % ) in all samples studied . The highest proportion of P28906 + cells costaining for c-kit was found in normal bone marrow ( 32.2 +/- 3.3 % ) . In normal peripheral blood and after cytokine mobilization , fewer of the P28906 + cells weakly expressed c-kit ( < 15 % ) . The low percentage and level of c-kit expression may indicate that the majority of cytokine-mobilized P28906 + cells are lineage-committed progenitor cells , as reflected by the coexpression pattern for P28907 , HLA-DR , and P20138 . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . Inhibition of granulocyte differentiation by P55008 cyclins D2 and D3 but not D1 . Growth factor-induced signals govern the expression of three D-type cyclins , which , in turn , function as regulatory subunits of cyclin-dependent kinases ( cdks ) to control cell cycle transitions during the late P55008 interval . 32D myeloid cells , which self-renew as uncommitted precursors in interleukin 3 ( P08700 ) , express cyclins D2 and D3 ( but not D1 ) in complexes with cdk4 and cdk2 . When transferred to granulocyte colony-stimulating factor ( DB00099 ) , 32D cells stop dividing and terminally differentiate to mature neutrophils . P12004 D and cdk4 expression ceased as cells underwent growth arrest in DB00099 , but cdk2 levels were sustained . 32D cells engineered to ectopically express D-type cyclins exhibited contracted P55008 intervals with a compensatory lengthening of S phase but remained P08700 dependent for cell growth ; those overexpressing cyclins D2 and D3 ( but not D1 ) were unable to differentiate and died in DB00099 . P12004 D2 mutants , which can not efficiently bind to , or functionally interact with , the retinoblastoma protein ( P06400 ) or its relatives ( P28749 ) did not block differentiation . Conversely , the introduction of a catalytically inactive cdk4 mutant into cells overexpressing cyclin D2 restored their G- P04141 response . The persistence of cdk2 and its predilection to functionally interact with cyclins D2 and D3 rather than D1 might explain the specificity of the differentiation blockade . Role of prostacyclin in the cardiovascular response to thromboxane A2 . Thromboxane ( Tx ) A2 is a vasoconstrictor and platelet agonist . DB00945 affords cardioprotection through inhibition of TxA2 formation by platelet cyclooxygenase ( P23219 ) . DB01240 ( DB01240 ) is a vasodilator that inhibits platelet function . Here we show that injury-induced vascular proliferation and platelet activation are enhanced in mice that are genetically deficient in the P43119 ( IP ) but are depressed in mice genetically deficient in the TxA2 receptor ( TP ) or treated with a TP antagonist . The augmented response to vascular injury was abolished in mice deficient in both receptors . Thus , DB01240 modulates platelet-vascular interactions in vivo and specifically limits the response to TxA2 . This interplay may help explain the adverse cardiovascular effects associated with selective P35354 inhibitors , which , unlike aspirin and nonsteroidal anti-inflammatory drugs ( NSAIDs ) , inhibit DB01240 but not TxA2 . DB01088 has potent anti-inflammatory properties on human monocyte-derived dendritic cells . BACKGROUND : The stable prostaglandin I2 analogue ( iloprost ) iloprost has been shown to inhibit allergic airway inflammation in mice by modulating the function of myeloid dendritic cells ( DCs ) . OBJECTIVE : The aim of the current study was to investigate the biological activity of iloprost on human monocyte-derived DCs . METHODS : I prostanoid ( IP ) receptor expression was analysed by RT-PCR . Cytokine secretion by DCs and P01730 + T cells was measured by ELISA . The expression of the transcription factor FoxP3 after co-culture of DCs with P01730 + CD45RA+ T cells was analysed by flow cytometry . RESULTS : Human monocyte-derived DCs were found to express mRNA specific for the P43119 IP , and stimulation with iloprost resulted in increased cyclic AMP levels in both immature DCs ( iDCs ) and mature DCs ( mDCs ) . Moreover , iloprost dose dependently inhibited the secretion of P01375 , P05231 , P10145 and IL-12p70 in mDCs , while it enhanced P22301 production . Changes in cytokine secretion were paralleled by an altered T-cell priming capacity of DCs : in co-culture experiments of iloprost-treated mDC and naïve CD45RA+ T cells , an induction of regulatory T cells could be observed , as demonstrated by increased intracellular FoxP3 expression and P22301 production . Additionally , iloprost inhibited the MIP-3beta-induced migration of mDCs . CONCLUSION : In summary , our results provide evidence that iloprost profoundly affects the function of human myeloid DCs . Therefore , iloprost might also be a new therapeutical option for the treatment of asthma in humans . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . DB01393 restores the inhibition of DB00094 -induced follicular development and steroidogenesis by tumor necrosis factor-alpha through peroxisome proliferator-activated receptor-gamma pathway in an in vitro mouse preantral follicle culture . We recently reported that bezafibrate , a lipid-lowering drug of the fibrate class , administered in addition to clomiphene citrate ( CC ) successfully induced ovulation in CC-resistant polycystic ovary syndrome ( PCOS ) patients . We hypothesized that bezafibrate may directly affect ovarian follicle development . P01308 resistance and compensatory hyperinsulinemia are important for the pathogenesis of PCOS . In this study , we first examined the effects of tumor necrosis factor-alpha ( P01375 ) , which plays a role in insulin resistance , on follicle development by using the follicle culture system . P01375 significantly inhibited follicle-stimulating hormone ( DB00094 ) -induced follicle development , 17beta-estradiol ( E2 ) secretion , and ovulation rate in a dose-dependent manner . We then examined whether bezafibrate treatment could rescue the inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 . DB01393 treatment rescued inhibition of follicle development , secretion of E2 , and ovulation rate by P01375 . We examined the expression of peroxisome proliferator-activated receptor ( Q07869 ) subtypes in mouse preantral follicles . As the protein expression of only P37231 was observed in mouse preantral follicles , we examined whether bezafibrate could affect follicle development and steroidogenesis through P37231 pathways . Treatment with GW1929 , a selective P37231 agonist , restored inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 , whereas treatment with GW9662 , a selective P37231 antagonist , canceled the restorative effects of bezafibrate . Collectively , the results in this study suggest that bezafibrate may directly exhibit a restorative effect on the inhibition of ovarian follicle development and steroidogenesis by P01375 through the P37231 pathway . 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 . [ Detection of a high-affinity prostaglandin I2 binding site in the human thyroid ] . This study is concerned with the identification and the pharmacological properties of P43119 binding sites on human thyroid membrane fractions . Scatchard analysis is not linear , revealing a high- and a low-affinity receptor binding site . ( 3 H ) DB01088 binding experiments were performed under various clinical conditions : in thyroid cancer the low-affinity binding sites disappear totally and the specific high-affinity binding sites are diminished according to the grade of differentiation of the cancer . An alteration in Bmax and Kd is also observed in cold nodules , in Hashimoto 's and Riedl 's thyroiditis and in hyperthyroidism , whereas hot nodules exhibit an increase in both the receptor subpopulations . The data provide evidence for specific DB01240 binding sites and support the suggestion of a direct regulatory key-role of DB01240 in thyroid intermediary metabolism . Lysophosphatidic acid accelerates the development of human mast cells . Mast cells ( MCs ) initiate immune responses from mucosal surfaces and perivascular spaces . P21583 ( P21583 ) regulates MC development and viability , but the role of innate serum factors in MC development is unexplored . Cultured cord blood-derived human MCs ( hMCs ) express mRNA transcripts for all 4 known receptors for lysophosphatidic acid ( P08519 ) , an abundant serum-associated lipid growth factor . In an P21583 -dependent serum-free culture system , P08519 ( 2.5-10 microM ) increased the total number of hMCs by approximately 10-fold compared with cultures maintained in the absence of P08519 under otherwise identical conditions . P08519 was comitogenic with P21583 but did not prolong MC survival . P08519 -mediated proliferation was blocked by VPC-32179 , a competitive antagonist of P08519 (1) and P08519 (3) receptors , and by pertussis toxin , and it was also attenuated by GW9662 , a selective antagonist of peroxisome proliferator-activated receptor ( Q07869 ) -gamma . P08519 accelerated the acquisition of hMC granules and increased Kit expression . hMCs derived in the presence of P08519 were functional , as evidenced by their immunoglobulin E ( IgE ) -dependent histamine release and by their characteristic proliferative responses to interleukin-3 ( P08700 ) , P05112 , and P15248 in combination with P21583 . Thus , P08519 acts through P08519 receptor and P37231 -dependent pathways to accelerate hMC proliferation and differentiation , and it modulates their phenotype without providing cytoprotection . P08519 could facilitate MC hyperplasia in inflammation associated with either innate or adaptive immunity . Sulfonylureas inhibit cytokine-induced eosinophil survival and activation . Eosinophils play a key role in the pathogenesis of asthma and other allergic inflammatory diseases . We have previously shown that treatment of eosinophils with lidocaine preferentially inhibits P05113 -induced survival . This inhibition can not be overcome by increasing concentrations of P05113 and is not due to the blocking of Na+ channels by lidocaine . Here we report that one class of K+ channel blockers , the sulfonylureas , inhibits eosinophil survival in a manner similar to lidocaine . The sulfonylurea glyburide inhibits eosinophil survival even at high concentrations of P05113 . In contrast , increasing concentrations of P08700 or granulocyte-macrophage P04141 overcome glyburide inhibition . Glyburide also blocks cytokine-induced eosinophil superoxide production . Similar results were seen with the sulfonylureas tolbutamide and glipizide . Interestingly , the effects of glyburide are not antagonized by the DB00171 -sensitive K+ channel openers cromakalim , pinacidil , or diazoxide . Although Scatchard analysis of [3H]glyburide binding to eosinophil membranes indicated that the high affinity sulfonylurea receptor ( Q09428 ) is not present on eosinophils , human eosinophils do express mRNA homologous to the sulfonylurea receptor family , in keeping with the presence of a sulfonylurea receptor . Finally , coculture of eosinophils with combinations of glyburide , lidocaine , and dexamethasone resulted in synergistic inhibition of cytokine-mediated eosinophil survival and superoxide production . These results have intriguing clinical implications for the treatment of eosinophil-associated diseases . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Cyclooxygenase inhibition abrogates aeroallergen-induced immune tolerance by suppressing prostaglandin I2 receptor signaling . BACKGROUND : The prevalence of allergic diseases has doubled in developed countries in the past several decades . Cyclooxygenase ( P36551 ) -inhibiting drugs augmented allergic diseases in mice by increasing allergic sensitization and memory immune responses . However , whether P36551 inhibition can promote allergic airway diseases by inhibiting immune tolerance is not known . OBJECTIVE : To determine the role of the P36551 pathway and prostaglandin I2 ( DB01240 ) signaling through the P43119 ( IP ) in aeroallergen-induced immune tolerance . METHODS : Wild-type ( WT ) BALB/c mice and IP knockout mice were aerosolized with ovalbumin ( OVA ) to induce immune tolerance prior to immune sensitization with an intraperitoneal injection of OVA/alum . The P36551 inhibitor indomethacin or vehicle was administered in drinking water to inhibit enzyme activity during the sensitization phase . Two weeks after sensitization , the mice were challenged with OVA aerosols . Mouse bronchoalveolar lavage fluid was harvested for cell counts and TH2 cytokine measurements . RESULTS : WT mice treated with indomethacin had greater numbers of total cells , eosinophils , and lymphocytes , and increased P05113 and P35225 protein expression in BAL fluid compared to vehicle-treated mice . Similarly , IP knockout mice had augmented inflammation and TH2 cytokine responses compared to WT mice . In contrast , the DB01240 analog cicaprost attenuated the anti-tolerance effect of P36551 inhibition . CONCLUSION : P36551 inhibition abrogated immune tolerance by suppressing DB01240 IP signaling , suggesting that DB01240 signaling promotes immune tolerance and that clinical use of P36551 -inhibiting drugs may increase the risk of developing allergic diseases . DB00501 enhances antigen-specific IgE and Th2 cytokine production . BACKGROUND : Treatment with anti-ulcer drugs has been shown to enhance IgE production against food antigens . However , little is known about the immunological effects of cimetidine , a histamine receptor type 2 ( P25021 ) antagonist that is widely used as an anti-ulcer drug , in allergy . Therefore , the present study investigated the role of cimetidine in Th2 immune responses in mice . METHODS : BALB/c mice were immunized intraperitoneally with ovalbumin ( OVA ) with and without cimetidine . The levels of cytokines in supernatants of spleen cells cultured in the presence of OVA for 4 days and the levels of total and OVA-specific IgG(1) , IgG(2a) and/or IgE in sera from these mice were determined by ELISA . RESULTS : Administration of cimetidine to OVA-sensitized BALB/c mice promoted Th2 cytokine secretion by OVA-stimulated spleen cells in vitro and increased serum levels of OVA-specific IgE , IgG(1) and IgG(2a) . CONCLUSIONS : These results indicate that cimetidine can enhance Th2 responses , suggesting that cimetidine may contribute to IgE production in allergies . The anticoagulant effect of PGI2S and tPA in transgenic umbilical vein endothelial cells is linked to up-regulation of PKA and PKC . The selection of vascular grafts for coronary artery bypass surgery is crucial for a positive outcome . This study aimed to establish a novel line of vascular endothelial cells with a potent anticoagulant effect . A lentiviral vector was used to stably transfect human umbilical vein endothelial cells ( HUVECs ) with PGI2S alone ( HUVEC-PGI2S ) or both PGI2S and tPA ( HUVEC-PGI2S-tPA ) . Both HUVEC-PGI2S and HUVEC-PGI2S-tPA cells over-expressing PGI2S and tPA were compared to mock-transfected cells . The enzyme-linked immuno sorbent assay ( ELISAs ) demonstrated that the anticoagulation components , P01008 and P00747 , were up-regulated and coagulation factor FVIII was down-regulated in both cell lines . QRT-PCR and western blotting demonstrated the vasodilation and platelet disaggregation proteins PKA , PKC , and P43119 were up-regulated in both cell lines , but MAPK expression was not altered in either cell line . However , cell viability and colony formation assays and cell cycle analysis demonstrated that both cell lines had a lower rate of cell growth and induced P55008 phase arrest . HUVEC-PGI2S and HUVEC-PGI2S-tPA cells have a potent anticoagulant effect and their use in vascular heterografts may decrease the risk of thrombosis . Basal cell carcinoma and variants in genes coding for immune response , DNA repair , folate and iron metabolism . Basal cell carcinoma ( BCC ) is one of the most common neoplasms in the world and its incidence has been increasing worldwide in recent years . BCCs are caused by an interplay between genetic and environment factors . We conducted a case-control association study in BCC patients and controls from Sweden and Finland . Fifteen single nucleotide polymorphisms ( SNPs ) , P05231 -174G/C , -634G/C , and -597G/A ; P22301 -1082G/A and -592C/A ; IL-1beta-511C/T ; NBS1 exon 5 Glu185Gln ; Q01831 exon 15 Lys939Gln ; P18074 exon 23 Lys751Gln ; P18887 exon 10 Arg399Gln ; O43542 exon 7 Thr241Met ; cyclin D1 exon 4 G870A ; P42898 exon 4 Ala222Val and exon 7 Glu429Ala ; Q30201 exon 4 C282Y were performed by Pyrosequencing and RFLP techniques . Most of the genotype distributions were in accordance with the Hardy-Weinberg equilibrium ( HWE ) , except for P22301 -1082G/A , where cases with BCC showed a significant deviation from HWE ( P = 0.04 ) . Linkage disequilibrium was observed between the -174 and -597 alleles in the P05231 gene in the present populations . No difference between BCC and controls appeared in any of the SNPs analyzed . Only the combined distributions of TT/AA genotypes in P42898 exon 4 ( C/T ) and exon 7 ( A/C ) showed slight increase in BCC compared to controls ( P < 0.07 , OR : 1.94 ; 95 % CI : 0.96-3.89 ) . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Cytokines and atherosclerosis : a comprehensive review of studies in mice . In the past few years , inflammation has emerged as a major driving force of atherosclerotic lesion development . It is now well-established that from early lesion to vulnerable plaque formation , numerous cellular and molecular inflammatory components participate in the disease process . The most prominent cells that invade in evolving lesions are monocyte-derived macrophages and T-lymphocytes . Both cell types produce a wide array of soluble inflammatory mediators ( cytokines , chemokines ) which are critically important in the initiation and perpetuation of the disease . This review summarizes the currently available information from mouse studies on the contribution of a specified group of cytokines expressed in atherosclerotic lesions , viz. interleukins ( IL-1 , P60568 , P08700 , P05112 , P05113 , P05231 , P22301 , IL-12 , Q14116 , IL-20 ) and macrophage-associated cytokines [ tumour necrosis factor-alpha ( P01375 ) ; macrophage migration inhibitory factor ( MIF ) ; interferon-gamma ( P01579 ) ; colony stimulating factors G- P04141 ,- P09603 ,-GM- P04141 ) to atherogenesis . Emphasis is put on the consistency of the effects of these cytokines , i.e. inasmuch an effect depends on the experimental approach applied ( overexpression/deletion , strain , gender , dietary conditions , and disease stage ) . An important outcome of this survey is ( i ) that only for a few cytokines there is sufficient consistent data allowing classifying them as typically proatherogenic ( IL-1 , IL-12 , Q14116 , MIF , P01579 , P01375 , and P09603 ) or antiatherogenic ( P22301 ) and ( ii ) that some cytokines ( P05112 , P05231 and GM- P04141 ) can exert pro- or anti-atherogenic effects depending on the experimental conditions . This knowledge can be used for improved early detection , prevention and treatment of atherosclerosis . P43119 up-regulates the expression of angiogenic genes in human endometrium via cross talk with epidermal growth factor Receptor and the extracellular signaling receptor kinase 1/2 pathway . DB01240 ( P06744 ) is a member of the prostanoid family of lipid mediators that mediates its effects through a seven-transmembrane G protein-coupled receptor ( IP receptor ) . Recent studies have ascertained a role for prostanoid-receptor signaling in angiogenesis . In this study we examined the temporal-spatial expression of the IP receptor within normal human endometrium and additionally explored the signaling pathways mediating the role of IP receptor in activation of target angiogenic genes . Quantitative RT-PCR analysis demonstrated the highest endometrial expression of the IP receptor during the menstrual phase compared with all other stages of the menstrual cycle . Immunohistochemical analysis localized the site of IP receptor expression to the glandular epithelial compartment with stromal and perivascular cell immunoreactivity . Expression of the immunoreactive IP receptor protein was greatest during the proliferative and early secretory phases of the menstrual cycle . To explore the role of the IP receptor in glandular epithelial cells , we used the Ishikawa endometrial epithelial cell line . Stimulation of Ishikawa cells and human endometrial biopsy explants with 100 nm iloprost ( a P06744 analog ) rapidly activated P27361 /2 signaling and induced the expression of proangiogenic genes , basic fibroblast growth factor , angiopoietin-1 , and angiopoietin-2 , in an epidermal growth factor receptor ( P00533 ) -dependent manner . Furthermore , P00533 colocalized with IP receptor in the glandular epithelial compartment . These data suggest that P06744 -IP interaction within glandular epithelial cells can promote the expression of proangiogenic genes in human endometrium via cross talk with the P00533 . Microglial activation , increased P01375 and P31645 expression in the prefrontal cortex define stress-altered behaviour in mice susceptible to anhedonia . A chronic stress paradigm comprising exposure to predation , tail suspension and restraint induces a depressive syndrome in C57BL/6J mice that occurs in some , but not all , animals . Here , we sought to extend our behavioural studies to investigate how susceptibility ( sucrose preference < 65 % ) or resilience ( sucrose preference > 65 % ) to stress-induced anhedonia affects the 5HT system and the expression of inflammation-related genes . All chronically stressed animals , displayed increased level of anxiety , but susceptible mice exhibited an increased propensity to float in the forced swim test and demonstrate hyperactivity under stressful lighting conditions . These changes were not present in resilient or acutely stressed animals . Compared to resilient animals , susceptible mice showed elevated expression of tumour necrosis factor alpha ( P01375 ) and the 5-HT transporter ( P31645 ) in the pre-frontal area . Enhanced expression of 5HT(2A) and P23219 in the pre-frontal area was observed in all stressed animals . In turn , indoleamine-2,3-dioxygenase ( P14902 ) was significantly unregulated in the raphe of susceptible animals . At the cellular level , increased numbers of Iba-1-positive microglial cells were also present in the prefrontal area of susceptible animals compared to resilient animals . Consequently , the susceptible animals display a unique molecular profile when compared to resilient , but anxious , animals . Unexpectedly , this altered profile provides a rationale for exploring anti-inflammatory , and possibly , P01375 -targeted therapy for major depression . Enhanced goblet cell hyperplasia in HDC knockout mice with allergic airway inflammation . BACKGROUND : DB11320 is known to have immunoregulatory roles in allergic reactions through histamine receptor 1 ( P35367 ) , P25021 , Q9Y5N1 and Q9H3N8 . However , its role in goblet cell hyperplasia in the airways of asthma patients is yet to be clarified . OBJECTIVE : This study was designed to examine the role of histamine in goblet cell hyperplasia using histamine-deficient mice ( Hdc-/- mice ) with allergic airway inflammation . METHODS : Wild-type and Hdc-/- C57BL/6 mice were sensitized with ovalbumin ( OVA ) . After a 2-week exposure to OVA , goblet cell hyperplasia was evaluated . Cell differentials and cytokines in BALF were analyzed . The mRNA levels of P98088 and Gob-5 gene were determined quantitatively . RESULTS : The number of eosinophils in BALF increased in both the sensitized wild-type mice and Hdc-/- mice with OVA inhalation . In addition , the numbers of alveolar macrophages and lymphocytes in BALF increased significantly in the sensitized Hdc-/- mice with OVA inhalation compared to the wild-type mice under the same conditions . The concentrations of P05112 ( P05112 ) , P05113 , P35225 , Interferon-gamma ( P01579 ) , tumor necrosis factor-alpha ( P01375 ) and P60568 in the BALF all increased significantly in both groups compared to those exposed to saline . In particular , the concentration of P01375 in the Hdc-/- mice exposed to OVA was significantly higher than that in the wild-type mice under the same conditions . The mRNA levels of Gob-5 and P98088 , and the ratio of the goblet cells in the airway epithelium significantly increased in Hdc-/- mice exposed to OVA compared to wild-type mice . CONCLUSIONS : These results suggested that histamine may play a regulatory role in goblet cell hyperplasia in allergic airway inflammation . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . P05362 -independent lymphocyte transmigration across high endothelium : differential up-regulation by interferon gamma , tumor necrosis factor-alpha and interleukin 1 beta . The adhesion of lymphocytes to cytokine-treated high endothelium was studied using cultured high endothelial cells ( O14777 ) . Pretreatment of the O14777 layer with a variety of cytokines caused up-regulation of lymphocyte adhesion with the effects ordered interferon gamma ( P01579 ) greater than tumor necrosis factor-alpha ( P01375 ) greater than or equal to interleukin 1 beta ( IL 1 beta ) . Increased lymphocyte adhesion was found to be independent of P05362 as expression by O14777 was not increased by cytokines and antibodies against P05362 did not block adhesion . The peptide CS1 and anti-beta 1 integrin subunit antibodies , however , caused partial inhibition of lymphocyte adhesion thus indicating a role for fibronectin on O14777 and alpha 4 beta 1 on lymphocytes . Study of the kinetics of lymphocyte adhesion showed that the effects of P01579 and P01375 were persistent and remained detectable 2.5 h after removal of the cytokines whereas the effects of IL 1 beta were transient and were not sustained beyond 1 h . All of the cytokines used caused transient increases in the number of surface-bound lymphocytes with P01579 greater than P01375 greater than or equal to IL 1 beta , however , the most dramatic effect was on the transmigration of lymphocytes across the O14777 . Both P01579 and P01375 caused sustained increased transmigration with P01579 having the greater effect . IL 1 beta had little effect on transmigration . This model demonstrates that the binding and transmigration of lymphocytes across O14777 can be differentially regulated by the actions of individual cytokines . These results support the concept that locally produced cytokines regulate O14777 function within the lymph node . Accelerated and long-term hematopoietic engraftment in mice transplanted with ex vivo expanded bone marrow . Using a murine experimental model , we investigated whether ex vivo expansion of BM grafts under P08700 / P05231 stimulation accelerates the early hematopoietic recovery of recipients of BM transplants . To facilitate the ex vivo expansion of hematopoietic progenitors , BM was first enriched in proliferatively active hematopoietic stem cells by a single treatment with 5-fluorouracil ( 5FU ) 4 days prior to the BM harvest . The results showed that the number of CFU-GM and CFU- P28222 progenitors in the graft was significantly increased ( 56-fold and 14-fold , respectively ) , as a result of a 3 day incubation in the presence of P08700 and P05231 . Daily analysis of animals transplanted with 5 x 10(4) BM cells , either freshly harvested or expanded for 3 days , showed that the expanded grafts consistently allowed a faster hematopoietic recovery of recipients . Differences between both groups of transplanted animals were most evident when the number of either femoral or splenic CFU-GMs were compared , with increases close to 70-fold at the fifth day of engraftment being observed . Similarly , mice transplanted with expanded grafts showed a hastened recovery in the cellularity of both organs that was most significant during the second week following transplantation , with maximal increases of 15 and 40-fold in the BM and spleen , respectively . Differences in peripheral leukocyte numbers between both groups of recipients were much less remarkable than those observed in the hematopoietic organs , although from the nadir period to the 11th day post-transplantation differences ranging from twofold to sixfold were apparent , consistent with a higher rate of mouse survival. ( ABSTRACT TRUNCATED AT 250 WORDS ) Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility .

MH_train_63

MH_train_63

MH_train_63

interacts_with DB00398?

multiple_choice

DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . The G protein-coupled P08908 receptor causes suppression of caspase-3 through MAPK and protein kinase Calpha . The 5-HT(1A) agonist 8-hydroxy-2 ( di-n-propylamino ) tetralin ( 8-OH-DPAT ) causes inhibition of caspase-3 and apoptosis via the extracellular signal-regulated kinases ( P27361 /2 ) in hippocampal P0CJ69 -5 cells . Two 5-HT(1A) agonists , Repinotan hydrochloride ( BAY x 3702 ) and 8-OH-DPAT , block caspase-3 activation and apoptosis caused by anoxia/reoxygenation and H(2)O(2) treatment . This is reversed upon transient expression of dominant negative Ras ( N17Ras ) and P04049 ( Raf301 ) , confirming the involvement of Ras and P04049 in this 5-HT(1A)-R --> P27361 /2 --> caspase-3 pathway . A selective inhibitor of phospholipase Cbeta ( PLCbeta ) ( U73122 ) but not a general protein kinase C ( PKC ) inhibitor ( GFX ) reversed the 5-HT(1A)-R-mediated P27361 /2 stimulation . However , both GFX and the PKCalpha and PKCbeta(1) inhibitor Gö6976 reversed the P27361 /2-mediated inhibition of caspase-3 . P29323 -dependent activation of only PKCalpha was observed in immunoprecipitates obtained from 5-HT(1A) agonist-treated P0CJ69 -5 cells . Finally , transient expression of kinase-negative PKCalpha eliminated the 8-OH-DPAT-evoked block on the H(2)O(2)-triggered caspase-3 stimulation , establishing PKCalpha as a link between P29323 and caspase-3 ( 5-HT(1A)-R --> P98160 --> P27361 /2 --> PKCalpha --> caspase-3 ) . Our results elucidate a novel yet general , neuroprotective pathway through which G protein-coupled receptors could cause inhibition of effector caspases , such as caspase-3 . Differential role of two P11473 coactivators , Q15648 and Q9Y6Q9 , in keratinocyte proliferation and differentiation . Cell programs such as proliferation and differentiation involve the selective activation and repression of gene expression . The vitamin D receptor ( P11473 ) , through 1,25(OH)(2)D(3) , controls the proliferation and differentiation of keratinocytes . Previously , we have identified two P11473 binding coactivator complexes . In proliferating keratinocytes P11473 bound preferentially to the DRIP complex , whereas in differentiated keratinocytes the P12931 complex was preferred . We proposed that different coactivators are required for sequential gene regulation in the transition from proliferation to differentiation . Here we examined the roles of Q15648 and Q9Y6Q9 in this transition . Silencing of Q15648 and P11473 caused hyperproliferation of keratinocytes , demonstrated by increased XTT and BrdU incorporation . Q9Y6Q9 silencing , on the other hand , did not have an effect on proliferation . In contrast , Q9Y6Q9 as well as Q15648 and P11473 silencing blocked keratinocyte differentiation as shown by decreased expression of keratin 1 and filaggrin . These results are consistent with the differential localization of Q15648 and Q9Y6Q9 in skin . These results indicate that Q15648 is required for keratinocyte proliferation . Both Q15648 and Q9Y6Q9 are required for the keratinocyte differentiation . These results support the concept that the selective use of coactivators by P11473 underlies the selective regulation of gene expression in keratinocyte proliferation and differentiation . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . DB00398 is tolerable and improves clinical outcomes in patients with P36888 -ITD acute myeloid leukemia prior to stem cell transplant and after relapse post-transplant . Hyperosmolar mannitol simulates expression of aquaporins 4 and 9 through a p38 mitogen-activated protein kinase-dependent pathway in rat astrocytes . The membrane pore proteins , aquaporins ( AQPs ) , facilitate the osmotically driven passage of water and , in some instances , small solutes . Under hyperosmotic conditions , the expression of some AQPs changes , and some studies have shown that the expression of P29972 and P55064 is regulated by MAPKs . However , the mechanisms regulating the expression of P55087 and AQP9 induced by hyperosmotic stress are poorly understood . In this study , we observed that hyperosmotic stress induced by mannitol increased the expression of P55087 and AQP9 in cultured rat astrocytes , and intraperitoneal infusion of mannitol increased P55087 and AQP9 in the rat brain cortex . In addition , a p38 MAPK inhibitor , but not P29323 and JNK inhibitors , suppressed their expression in cultured astrocytes . AQPs play important roles in maintaining brain homeostasis . The expression of P55087 and AQP9 in astrocytes changes after brain ischemia or traumatic injury , and some studies have shown that p38 MAPK in astrocytes is activated under similar conditions . Since mannitol is commonly used to reduce brain edema , understanding the regulation of AQPs and p38 MAPK in astrocytes under hyperosmotic conditions induced with mannitol may lead to a control of water movements and a new treatment for brain edema . Multi-kinase inhibition in ovarian cancer . DB00398 ( Nexavar ) is a multi-kinase inhibitor that was developed as an inhibitor of RAF-1 , in the P27361 /2 pathway , but which was subsequently shown to inhibit class III tyrosine kinase receptors . ( 1 ) More recently regorafenib ( Stivarga ) has been developed , which is a further fluorinated version of sorafenib with greater bioavailability and similar inhibitory properties against RAF-1/class III RTKs . ( 2 ) Some of the anti-tumor effects of sorafenib have been ascribed to anti-angiogenic actions of this agent on endothelial associated kinases such as P35968 . Other effects of sorafenib clearly have to be due to its effects on the inherent biology of the tumor cells themselves . For example , through various mechanisms sorafenib has been shown in the laboratory and the clinic to suppress expression of the protective protein Q8WXI8 -1 . ( 3 ) DB00398 has also been linked to inhibition of P40763 , NFκB , and activation of the death receptor CD95 . ( 4 ) DB00398 is routinely dosed daily ( 400 mg P55957 ) and 7 d after the start of dosing has a Cmax of ~21 μM with a nadir at 12 h of ~10 μM , and is a highly protein bound based on in vitro assays . ( 5 ) Despite this in vitro binding data sorafenib has profound in vivo effects on tumor cells in renal carcinoma and hepatocellular carcinoma patients ; cells which are not per se addicted to high activity oncogene signals that are targets of sorafenib/regorafenib . Thus the precise stable bioavailable level of sorafenib/regorafenib in patient plasma is not known . DB00398 -associated remission of psoriasis in hypernephroma : case report . Psoriasis is a disease characterized by epidermal hyperproliferation that results in the formation of lesional plaques covered by scale . Psoriasis is thought to be angiogenesis dependent . Clear cell renal cell carcinoma is a hypervascularized solid tumor associated with loss of function of the von Hippel-Lindau ( P40337 ) tumor suppressor gene and increased P04049 activity . A 68-year-old man who suffered from recalcitrant psoriasis for over 50 years was treated with sorafenib for metastatic clear cell renal carcinoma . One month later , his psoriasis , previously 8 x 6 cm on the mid posterior thorax , completely resolved . DB00398 works by inhibiting several receptor tyrosine kinases ( RTKs ) , such as vascular endothelial growth factor ( VEGFR ) and platelet-derived growth factor receptor ( P09619 ) ) . It also inhibits intracellular Raf kinase ( P04049 ) , which targets the ubiquitous mitogen-activated protein kinase ( MAPK ) intracellular signal transduction pathway . We suggest that this patient 's remission of psoriasis could be related to the inhibition/modulation of P15692 , P09619 , P04049 , and MAPK . DB00398 enhances pemetrexed cytotoxicity through an autophagy-dependent mechanism in cancer cells . DB00642 ( ALIMTA ) is a folate anti-metabolite that has been approved for the treatment of non-small cell lung cancer , and has been shown to stimulate autophagy . In the present study , we sought to further understand the role of autophagy in the response to pemetrexed and to test if combination therapy could enhance the level of toxicity through altered autophagy in tumor cells . The multikinase inhibitor sorafenib ( NEXAVAR ) , used in the treatment of renal and hepatocellular carcinoma , suppresses tumor angiogenesis and promotes autophagy in tumor cells . We found that sorafenib interacted in a greater than additive fashion with pemetrexed to increase autophagy and to kill a diverse array of tumor cell types . Tumor cell types that displayed high levels of cell killing after combination treatment showed elevated levels of AKT , P08133 S6K and/or phosphorylated P42345 , in addition to class III RTKs such as PDGFRb and P17948 , known in vivo targets of sorafenib . In xenograft and in syngeneic animal models of mammary carcinoma and glioblastoma , the combination of sorafenib and pemetrexed suppressed tumor growth without deleterious effects on normal tissues or animal body mass . Taken together , the data suggest that premexetred and sorafenib act synergistically to enhance tumor killing via the promotion of a toxic form of autophagy that leads to activation of the intrinsic apoptosis pathway , and predict that combination treatment represents a future therapeutic option in the treatment of solid tumors . DB00398 inhibits nuclear factor kappa B , decreases inducible nitric oxide synthase and cyclooxygenase-2 expression , and restores working memory in APPswe mice . Alzheimer 's disease ( AD ) is characterized by memory loss and the upregulation of pro-neuroinflammatory factors such as P04049 -1 , cyclooxygenase-2 ( Cox-2 ) , and the nuclear factor kappa B ( NF-kappaB ) , as well as a downregulation of protein kinase A ( PKA ) activity and the activation by phosphorylation of its downstream factor CREB . We investigated the effect of the anti-cancer P04049 -1 inhibitor , sorafenib tosylate ( Nexavar ) , on the expression of these factors and on the cognitive performance of aged APPswe mice . We found that chronic treatment with sorafenib stimulated PKA and CREB phosphorylation and inhibited P04049 -1 and NF-kappaB in the brains of APPswe mice . NF-kappaB controls the expression of several genes related to AD pathology , including P35228 and Cox-(2)Concurrent with NF-kappaB inhibition , sorafenib treatment decreased the cerebral expression of Cox-2 and P35228 in APPswe mice . It has recently been observed that Cox-2 inhibition prevents cognitive impairment in a mouse model of AD and amyloid beta peptide ( Abeta ) -induced inhibition of long-term potentiation ( LTP ) . Consistent with the idea that Cox-2 inhibition can improve cognitive abilities , we found that sorafenib restored working memory abilities in aged APPswe mice without reducing Abeta levels in the brain . These findings suggest that sorafenib reduced AD pathology by reducing neuroinflammation . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . P15692 differentially activates P40763 in microvascular endothelial cells . Increased P15692 expression is found in several pathologies characterized by abnormal angiogenesis . Previous studies have shown that the transcription factor P40763 mediates P15692 gene transcription and its activation . In this study , Western analysis and confocal immunocytochemistry were used to examine P40763 activation in retinal microvascular endothelial cells ( BREC ) . We found that P15692 rapidly induces P40763 tyrosine phosphorylation and nuclear translocation . Immunoprecipitation studies also showed that P15692 forms a complex with P35968 only in BREC and not in aortic macrovascular endothelial cells ( BAEC ) . In addition , quantitative real-time RT-PCR analysis of P15692 -induced P15692 expression showed a significant increase in specific mRNA formation only in BREC and not in BAEC , and this effect was significantly reduced by antisense-mediated reduction of P40763 expression . Furthermore , studies conducted in human dermal microvascular endothelial cells ( HDMEC ) showed that , in this endothelial cell type , P15692 autocrine expression is also accompanied by P40763 activation as in BREC . In this study we showed that P15692 can differentially induce P40763 activation in micro- versus macro-vascular endothelial cells and that this effect is linked to P35968 / P40763 complex formation , which correlates with P15692 autocrine ability to stimulate its own gene expression . P36888 -ITD knockin impairs hematopoietic stem cell quiescence/homeostasis , leading to myeloproliferative neoplasm . Internal tandem duplication ( ITD ) mutations within the P07333 -like tyrosine kinase-3 ( P36888 ) render the receptor constitutively active driving proliferation and survival in leukemic blasts . Expression of P36888 -ITD from the endogenous promoter in a murine knockin model results in progenitor expansion and a myeloproliferative neoplasm . In this study , we show that this expansion begins with overproliferation within a compartment of normally quiescent long-term hematopoietic stem cells ( LT-HSCs ) , which become rapidly depleted . This depletion is reversible upon treatment with the small molecule inhibitor DB00398 , which also ablates the disease . Although the normal LT- P19526 has been defined as P36888 (-) by flow cytometric detection , we demonstrate that P36888 is capable of playing a role within this compartment by examining the effects of constitutively activated P36888 -ITD . This indicates an important link between stem cell quiescence/homeostasis and myeloproliferative disease while also giving novel insight into the emergence of P36888 -ITD mutations in the evolution of leukemic transformation . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Abnormalities of Q9H3D4 , p53 , P36888 and N-ras genes and their prognostic value in relapsed acute myeloid leukemia . To clarify the role of the genetic mutations in the clinical course of acute myeloid leukemias ( AML ) , we analyzed for Q9H3D4 , p53 , P36888 and N-ras gene mutations and the expression of the Q9H3D4 gene in relapsed AML . Paired samples obtained from patients with AML at both stages of diagnosis and first relapse were analyzed . Twenty-four patients with relapsed AML survived for 6 to 81 months ( median 24 months ) from diagnosis . In one patient , no point mutation of the Q9H3D4 gene was detected , but loss of the Q9H3D4 gene expression was observed at both stages . Point mutations of the p53 gene were positive at both stages ( +/+ ) in two patients and negative at diagnosis and positive at relapse ( -/+ ) in two patients . Tandem duplication of the P36888 gene was detected in five patients at both stages ( +/+ ) . N-ras gene mutations at both stages ( +/+ ) were detected in three patients . Mutant p53 at relapse was associated with short survival in patients with relapsed AML ( P < 0.014 ) . Our findings show that p53 mutations were , at least in part , associated with the mechanism of relapse in AML , while new Q9H3D4 , P36888 and N-ras gene alterations did not occur at relapse . Loss of the Q9H3D4 gene expression in de novo AML has not been reported yet . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . [ P35354 inhibitor celecoxib can suppress the proliferation of P36888 -ITD positive acute myeloid leukemia cells with prominent down regulation of MEK/ Q8WXI8 -1 expression in vitro ] . The purpose of this study was to investigate the effects of Celecoxib on the proliferation of the P36888 -ITD positive and negative acute myeloid leukemia cells and its mechanism . The proliferation inhibition effect of Celecoxib with different doses on the P36888 -ITD positive cells MV4-11 and the P36888 -ITD negative K562 cells was detected by CCK-8 method , the cell apoptosis was determined by flow cytometry , and the MEK , Mcl-1 , pAKT expression was tested by Western blot . The results showed that Celecoxib inhibited the proliferation of both MV4-11 and K562 cells , but the IC50 for MV4-11 was ( 29.14 ± 2.4 ) µmol/L , which was significantly lower than that of K562 cells ( 39.84 ± 1.0 ) µmol/L ( P < 0.05 ) ; The induced apoptosis rate of Celecoxib at 20-80 µmol/L on MV4-11 was not observed , but there was apparent influence on K562 at the same concentration . Western blot showed that Celecoxib down-regulated the expression of MEK and Mcl-1 but did not change the expression of pAKT obviously on MV4-11 cells , while the expression of Mcl-1 was reduced a little , but no obvious change were found in the expression of MEK and pAKT on K562 cells . It is concluded that the Celecoxib can inhibit the proliferation of P36888 -ITD positive AML cells distinctly , and the potential mechanism may be related to the inhibition of the MEK/Mcl-1 signaling pathway . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Molecular mechanisms and modulation of key pathways underlying the synergistic interaction of sorafenib with erlotinib in non-small-cell-lung cancer ( NSCLC ) cells . Combination of drugs with different targets is a logical approach to overcome multilevel cross-stimulation among key pathways in NSCLC progression such as P00533 , K-Ras and VEGFR . The sorafenib-erlotinib combination showed clinical activity and acceptable safety . Therefore , we evaluated mechanisms underlying sorafenib-erlotinib interaction in seven NSCLC cell lines selected for their heterogeneous pattern of P00533 and Raf-kinase-inhibitor protein ( P30086 ) expression , and P00533 /K-Ras mutations . Pharmacologic interaction was studied using MTT/ P50991 assays and the combination index ( CI ) method , while effects on P00533 , Erk1/2 and Akt phosphorylation , cell cycle and apoptosis were studied with western-blot , ELISA , and flow cytometry . Intracellular drug concentrations were measured with LC-MS/MS , whereas kinase activity profiles were generated on tyrosine kinase peptide substrate arrays . Synergism was detected in all cell lines , with CIs < 0.6 in K-Ras mutated A549 , SW1573 and H460 , as well as in H1975 ( P00533 -T790M ) cells . DB00398 slowed cell cycle progression and induced apoptosis , which was significantly increased in the combination . Moreover , sorafenib reduced Akt/ P29323 phosphorylation in erlotinib-resistant cells , associated with significant P30086 up-regulation . No direct drug interaction was detected by LC-MS/MS measurement , while lysates from A549 and H1975 cells exposed to erlotinib+sorafenib showed a significant inhibition in the phosphorylation of 16 overlapping peptides , including sites from RAF , P35968 , P09619 , P24941 and P12931 , suggesting new markers to identify NSCLC patients who are likely to respond to this treatment . In conclusion , several mechanisms , including apoptosis-induction , modulation of expression/phosphorylation of P30086 and crucial kinases contribute to erlotinib-sorafenib synergistic interaction and should be evaluated in future trials for the rational development of this combination in NSCLC . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . Role of sorafenib in the treatment of advanced hepatocellular carcinoma : An update . DB00398 is the first and only p.o. administrated drug currently approved to treat advanced hepatocellular carcinoma ( HCC ) . However , concerns have been raised about sorafenib therapy , including acquired drug resistance . This review provides an overview of sorafenib in the treatment of HCC on the basis of data obtained in the laboratory and in clinical studies . Three underlying mechanisms have been found to support sorafenib therapy . First , sorafenib blocks HCC cell proliferation by inhibiting BRaf and Raf1/c-Raf serine/threonine kinase phosphorylation in the mitogen-activated protein kinase pathway . Second , sorafenib induces apoptosis by reducing elF4E phosphorylation and downregulating Mcl-1 levels in tumor cells . Third , sorafenib prevents tumor-associated angiogenesis by inactivating vascular endothelial growth factor receptors ( P35968 and -3 ) and the platelet-derived growth factor receptor-β . Clinical trials have demonstrated the effectiveness and relative safety of sorafenib , and thus the drug is used in unresectable HCC . However , many patients may develop acquired resistance to sorafenib , so their response to sorafenib is eventually lost . DB00398 may induce autophagy , which leads to apoptosis . However , autophagy can also cause drug resistance . Many studies have combined sorafenib with other treatments in an effort to increase its effects , reduce the necessary dose or overcome resistance . It is urgent to study the mechanisms underlying how sorafenib interacts with cellular molecules and other drugs to increase its efficacy and reduce resistance in HCC patients . Aripiprazole : pharmacodynamics of a dopamine partial agonist for the treatment of schizophrenia . Aripiprazole is the first approved atypical antipsychotic with a mechanism of action that exerts a partial agonism with high affinity at DB00988 D2- and Serotonin- P08908 -receptors as well as an antagonism at Serotonin-5- Q13049 -receptors . Aripiprazole provides good clinical effectiveness and a favorable profile of safety and tolerability . The special pharmacodynamics of aripiprazole are described herein . Inhibitors of P11274 signalling interrupt the survival signal mediated by the micro-environment in mantle cell lymphoma . Several studies provide evidences for mantle cell lymphoma ( Q8WXI8 ) cell survival relying on B-cell receptor ( P11274 ) -mediated signalling pathways , whereas the nature of this activation is unknown . Significant progress in Q8WXI8 treatment is achieved through therapies targeting P11274 -associated kinases , i.e. , DB09053 and Fostamatinib , inhibitors of Q06187 and P43405 , respectively . Our study addresses survival signals emanating from the P11274 or the tumour environment and how inhibiting P11274 signalling effectors might impact these survival signals . We found that Q06187 was constitutively activated and that P43405 phosphorylation was highly increased and sustained upon P11274 activation of primary Q8WXI8 cells . Moreover , Q8WXI8 cells from leukaemic patients secreted high amount of IL-1β , P05231 , P10145 and P13501 . Activation of the P11274 induced ( i ) cell survival , ( ii ) P40763 activation and ( iii ) increased autocrine secretion of IL-1β , P05231 , P10145 , P13501 , P22301 , TNFα and P15692 . Specific inhibition of Q06187 by DB09053 or P43405 by Fostamatinib ( R406 ) reversed these protective effects and decreased both basal and P11274 -induced autocrine cytokine secretions associated with P40763 phosphorylation . Interestingly , targeting Q06187 and P43405 prevented and inhibited P11274 -induced Q8WXI8 cell adhesion to human bone marrow stromal cells ( HMSCs ) in short- and long-term co-culture . We demonstrated that P11274 -induced survival relies on autocrine secretion of IL-1β , TNFα and P13501 that might facilitate adhesion of Q8WXI8 cells to HMSC . Treatment with DB09053 or Fostamatinib blocked the chemotactic signal thus increasing apoptosis . DB00184 activates P46937 through nAChRs mediated signaling in esophageal squamous cell cancer ( ESCC ) . Cigarette smoking is an established risk factor for esophageal cancers . P46937 ( P46937 ) , the key transcription factor of the mammalian Hippo pathway , has been reported to be an oncogenic factor for many cancers . In this study , we find nicotine administration can induce nuclear translocation and activation of P46937 in ESCC . Consistently , we observed nuclear translocation and activation of P46937 by knockdown of P32297 , which is a negative regulator of nicotine signaling in bronchial and esophageal cancer cells . DB00184 administration or P32297 depletion substantially increased proliferation and migration in esophageal cancer cells . Interestingly , we find that P46937 physically interacts with nAChRs , and nAChRs-signaling dissociates P46937 from its negative regulatory complex composed with α-catenin , β-catenin and 14-3-3 in the cytoplasm , leading to upregulation and nuclear translocation of P46937 . This process likely requires PKC activation , as PKC specific inhibitor Enzastaurin can block nicotine induced P46937 activation . In addition , we find nicotine signaling also inhibits the interaction of P46937 with Q9H3D4 , which contributes to the inhibitory effect of nicotine on apoptosis . Using immunohistochemistry analysis we observed upregulation of P46937 in a significant portion of esophageal cancer samples . Consistently , we have found a significant association between P46937 upregulation and cigarette smoking in the clinical esophageal cancer samples . Together , these findings suggest that the nicotine activated nAChRs signaling pathway which further activates P46937 plays an important role in the development of esophageal cancer , and this mechanism may be of a general significance for the carcinogenesis of smoking related cancers . DB00398 ( BAY 43-9006 ) in hepatocellular carcinoma patients : from discovery to clinical development . Angiogenesis and signaling through the DB01367 /RAF/mitogen-activated protein/extracellular signal-regulated kinase ( P29323 ) kinase ( MEK ) / P29323 cascade have been reported to play important roles in the development of hepatocellular carcinoma ( HCC ) . DB00398 ( Nexavar ) , a novel bi-aryl urea BAY 43-9006 , is an orally administered multikinase inhibitor with activity against DB01367 /RAF kinases multikinase inhibitor with activity against RAF kinases and several receptor tyrosine kinases , including vascular endothelial growth factor receptor ( VEGFR ) , platelet-derived growth factor receptor ( P09619 ) , P36888 , Ret , and c-Kit . It is involved in angiogenic pathway and cell proliferation . DB00398 has demonstrated potent anti-tumor activity in in vitro studies , preclinical xenograft models of different tumor types and human clinical trials . This review summarizes the history of sorafenib from its discovery by the medicinal chemistry approach through to clinical development and ongoing trials on the combination between sorafenib and trans-arterial chemoembolization ( P78536 ) in HCC patients . Targeted therapy in gastric cancer . Gastric cancer is often diagnosed at an advanced stage . Although chemotherapy prolongs survival and improves quality of life , the survival of gastric cancer patients with advanced disease is short . Thanks to recent insights into the molecular pathways involved in gastric carcinogenesis , new targeted treatment options have become available for gastric cancer patients . DB00072 , an antibody targeted to HER-2 , was shown to improve survival of advanced gastric cancer patients harboring HER-2 overexpression due to gene amplification in their tumor cells , and is currently also explored in adjuvant and neoadjuvant settings . Another agent with promising results in clinical trials is ramucirumab , an antibody targeting P35968 . No clear survival benefit , however , were experienced with agents targeting P00533 ( cetuximab , panitumumab ) , P15692 ( bevacizumab ) , or P42345 ( everolimus ) . Drugs targeting c-MET/ P14210 are currently under investigation in biomarker-selected cohorts , with promising results in early clinical trials . This review will summarize the current status of targeted treatment options in gastric cancer .

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[ Low doses of sulphonyluria as a successful replacement for insulin therapy in a patient with neonatal diabetes due to a mutation of Q14654 gene encoding Kir6.2 ] . Neonatal diabetes mellitus is a rare metabolic disorder with an estimated incidence of 1:300.000 to 400.000 newborns , and less than 50 % of the neonates have permanent neonatal diabetes mellitus ( PNDM ) . Recently , activating mutation in the Q14654 gene encoding Kir6.2 subunit of the adenosin triphosphate-sensitive potassium ( K( DB00171 ) ) channel has been described as the most frequent cause of PNDM . Under physiological circumstances K( DB00171 ) channel closure plays a central role in glucose-stimulated insulin secretion from pancreatic beta cells . Sulphonylurea drugs stimulate insulin secretion by binding to and closing K( DB00171 ) channels and thus bypassing beta cell metabolism stimulate the same chain of reactions as glucose . We describe a boy diagnosed with PNDM at the age of 3 months when insulin therapy was started , and at the age of 4.5 years Q14654 gene was sequenced and found that the boy carried a de novo activating R201H mutation . P01308 therapy was successfully switched to low doses of oral glibenclamide . Accordingly , it is important to emphasize that every person diagnosed with diabetes before six months of life , however old they actually are , should be tested for K( DB00171 ) mutations which is offered via the website www.diabetesgenes.org . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . Prolyl-4-hydroxylase domain protein 2 controls NF-κB/p65 transactivation and enhances the catabolic effects of inflammatory cytokines on cells of the nucleus pulposus . Prolyl-4-hydroxylase ( P20941 ) proteins are key in sensing tissue hypoxia . In nucleus pulposus ( NP ) cells , our previous work demonstrated that P20941 isoforms have a differential contribution in controlling hypoxia-inducible factor ( HIF ) -α degradation and activity . Recently we have shown that a regulatory relationship exists between Q9H6Z9 and inflammatory cytokines in NP cells . With respect to Q9GZT9 , the most abundant P20941 isoform in NP cells , very little is known concerning its function and regulation under inflammatory conditions that characterize intervertebral disc degeneration . Here , we show that Q9GZT9 is a potent regulator of the catabolic activities of P01375 -α ; silencing of Q9GZT9 significantly decreased P01375 -α-induced expression of catabolic markers including P31431 , P08254 , P45452 , and Q9UNA0 , as well as several inflammatory cytokines and chemokines , while partially restoring aggrecan and collagen II expression . Use of NF-κB reporters with ShPHD2 , SiHIF-1α , as well as p65(-/-) , Q9GZT9 (-/-) , and Q9H6Z9 (-/-) cells , shows that Q9GZT9 serves as a co-activator of NF-κB/p65 signaling in Q9BYW2 -independent fashion . Immunoprecipitation of endogenous and exogenously expressed tagged proteins , as well as fluorescence microscopy , indicates that following P01375 -α treatment , Q9GZT9 interacts and co-localizes with p65 . Conversely , loss of function experiments using lentivirally delivered Sh-p65 , Sh-IKKβ , and NF-κB inhibitor confirmed that cytokine-dependent Q9GZT9 expression in NP cells requires NF-κB signaling . These findings clearly demonstrate that Q9GZT9 forms a regulatory circuit with P01375 -α via NF-κB and thereby plays an important role in enhancing activity of this cytokine . We propose that during disc degeneration Q9GZT9 may offer a therapeutic target to mitigate the deleterious actions of P01375 -α , a key proinflammatory cytokine . [ Anti-cholesterol agents , new therapeutic approaches ] . Statins and fibrates constitute the two major families of lipid-lowering agents . Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia . Both drugs are also used for the treatment of mixed dyslipidemia . Some fibrates efficiently lower serum LDL-cholesterol . Statins inhibit P04035 and decrease cellular cholesterol synthesis . The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the P01130 gene . This over expression of the P01130 in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels . The effects of fibrates on lipid metabolism are entirely due to their capacity to activate Q07869 and to induce the over expression of genes containing a PPRE in their promoter . Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis . Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription , respectively . Fibrates could decrease triglycerides partly by inducing apo A-V over-expression . These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription . Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism . A lactate-induced response to hypoxia . Organisms must be able to respond to low oxygen in a number of homeostatic and pathological contexts . Regulation of hypoxic responses via the hypoxia-inducible factor ( HIF ) is well established , but evidence indicates that other , HIF-independent mechanisms are also involved . Here , we report a hypoxic response that depends on the accumulation of lactate , a metabolite whose production increases in hypoxic conditions . We find that the Q9UGV2 protein is degraded in a Q9GZT9 / P40337 -dependent manner in normoxia but is protected from destruction by binding to lactate that accumulates under hypoxia . The stabilized Q9UGV2 protein binds c-Raf to mediate hypoxia-induced activation of Raf- P29323 pathway , promoting angiogenesis and cell growth . Inhibiting cellular lactate production abolishes the Q9UGV2 -mediated hypoxia responses . Our study , therefore , elucidates the molecular basis for lactate-induced hypoxia signaling , which can be exploited for the development of therapies targeting hypoxia-induced diseases . Beta-cell alpha-ketoglutarate hydroxylases may acutely participate in insulin secretion . The presence of Fe(II) alpha-ketoglutarate hydroxylases in rat and human pancreatic islets and P01308 -1 832/13 cells was demonstrated with the reverse transcriptase polymerase chain reaction ( Q96KS0 , 2 , and 3 ; lysyl hydroxylases 1 , 2 , and 3 ; and phytanoyl-coenzyme A hydroxylase were seen ) and/or immunoblotting ( high levels of proline hydroxylase P4Halpha1 , Q9GZT9 , and PHD4 and low levels of Q9GZT9 and Q9H6Z9 in human islets , and high levels of Q9GZT9 in rat islets and P01308 -1 cells were seen ) . Prolyl hydroxylase enzyme activity in P01308 -1 832/13 cells was purified with polyproline affinity chromatography . Inhibitors of alpha-ketoglutarate hydroxylases lowered glucose-induced and leucine-plus-glutamine-induced insulin release in rat pancreatic islets , suggesting that there may be acute unknown effects of alpha-ketoglutarate hydroxylases in insulin secretion . It is possible that an increase in mitochondrially generated alpha-ketoglutarate derived from insulin secretagogue carbon and translocated to the cytosol may be part of the signal for insulin secretion . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . A study of 36 unrelated cases with pure erythrocytosis revealed three new mutations in the erythropoietin receptor gene . Thirty-six unrelated cases with erythrocytosis of unknown origin were investigated . Exons 5-8 of the erythropoietin receptor gene ( P19235 ) , the von Hippel-Lindau gene , and the prolyl hydroxylase domain protein 2 gene ( Q9GZT9 ) were screened by direct DNA sequencing . The O60674 mutation , O60674 ( Val617Phe ) , was screened by allele specific PCR . In this study , three new mutations of P19235 causing deletions in exon 8 were found : the first led directly to a stop codon [ g.5957_5958delTT ( p.Phe424X ) ] , the second to a stop codon after one residue [ g.5828_5829delCC ( p.Pro381GlnfsX1 ) ] and the third to a stop codon following a frameshift sequence of 23 residues [ g.5971delC ( p.Leu429TrpfsX23 ) ] . One patient had a previously reported P19235 mutation [ g.6146A > G ( p.Asn487Ser ) ] and another , a silent one ( g.5799G > A ) . All were heterozygotes . In addition , 2 patients were positive for O60674 ( Val617Phe ) , and 2 reported elsewhere , were mutated in the Q9GZT9 gene [ c.606delG ( p.Met202IlefsX71 ) . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Genetic determinants of Tibetan high-altitude adaptation . Some highland populations have genetic adaptations that enable their successful existence in a hypoxic environment . Tibetans are protected against many of the harmful responses exhibited by non-adapted populations upon exposure to severe hypoxia , including elevated hemoglobin concentration ( i.e. , polycythemia ) . Recent studies have highlighted several genes subject to natural selection in native high-altitude Tibetans . Three of these genes , Q99814 , Q9GZT9 and Q07869 , regulate or are regulated by hypoxia inducible factor , a principal controller of erythropoiesis and other organismal functions . Uncovering the molecular basis of hypoxic adaptation should have implications for understanding hematological and other adaptations involved in hypoxia tolerance . Because the hypoxia response involves a variety of cardiovascular , pulmonary and metabolic functions , this knowledge would improve our understanding of disease mechanisms and could ultimately be translated into targeted therapies for oxygen deprivation , cardiopulmonary and cerebral pathologies , and metabolic disorders such as diabetes and obesity . Kinin-B2 receptor exerted neuroprotection after diisopropylfluorophosphate-induced neuronal damage . The kinin-B2 receptor ( B2BKR ) activated by its endogenous ligand bradykinin participates in various metabolic processes including the control of arterial pressure and inflammation . Recently , functions for this receptor in brain development and protection against glutamate-provoked excitotoxicity have been proposed . Here , we report neuroprotective properties for bradykinin against organophosphate poisoning using acute hippocampal slices as an in vitro model . Following slice perfusion for 10min with diisopropylfluorophosphate ( DB00677 ) to initiate the noxious stimulus , responses of pyramidal neurons upon an electric impulse were reduced to less than 30 % of control amplitudes . Effects on synaptic-elicited population spikes were reverted when preparations had been exposed to bradykinin 30min after challenging with DB00677 . Accordingly , bradykinin-induced population spike recovery was abolished by HOE-140 , a B2BKR antagonist . However , the kinin-B1 receptor ( B1BKR ) agonist Lys-des- DB00125 (9)-bradykinin , inducing the phosphorylation of mitogen-activated protein kinase ( MEK/MAPK ) and cell death , abolished bradykinin-mediated neuroprotection , an effect , which was reverted by the P29323 inhibitor PD98059 . In agreement with pivotal B1BKR functions in this process , antagonism of endogenous B1BKR activity alone was enough for restoring population spike activity . On the other hand pralidoxime , an oxime , reactivating acetylcholinesterase ( P22303 ) after organophosphate poisoning , induced population spike recovery after DB00677 exposure in the presence of bradykinin and Lys-des- DB00125 (9)-bradykinin . Lys-des- DB00125 (9)-bradykinin did not revert protection exerted by pralidoxime , however when instead bradykinin and Ly-des- DB00125 (9)-bradykinin were superfused together , recovery of population spikes diminished . These findings again confirm the neuroprotective feature of bradykinin , which is , diminished by its endogenous metabolites , stimulating the B1BKR , providing a novel understanding of the physiological roles of these receptors . Disparate effects of simvastatin on angiogenesis during hypoxia and inflammation . AIMS : Studies have shown that some of statin 's pleiotropic effects were achieved by either promotion or inhibition of angiogenesis , depending on the underlying disease . This study tested the hypothesis that the angiogenic potential of simvastatin is related to the microenvironmental conditions . MAIN METHODS : Human umbilical vein endothelial cells ( HUVEC ) were studied after exposure to hypoxia or the inflammatory factors tumor necrosis factor ( P01375 ) -alpha , with or without co-incubation with simvastatin ( 1 micromol/L ) and mevalonate . HUVEC angiogenesis was evaluated by tube formation , migration , and proliferation assays . Hypoxia inducible factor ( HIF ) -1alpha , vascular endothelial growth factor ( P15692 ) , Akt , endothelium nitric oxide synthase ( e-NOS ) , and oxidative stress were evaluated . KEY FINDINGS : HUVEC angiogenesis increased during hypoxia ( tube length 14.7+/-0.5 vs. 7.8+/-0.6 mm , p < 0.05 ) and further enhanced by simvastatin ( 19.3+/-1.1 mm , p < 0.05 vs. hypoxia alone ) , which downregulated the expression of the Q9BYW2 inhibitor Q9GZT9 and upregulated HIF-1alpha , P15692 , and Akt , without changing oxidative stress or P29474 . Incubation with P01375 promoted HUVEC angiogenesis ( 7.4+/-0.2 vs. 6.5+/-0.2 mm , p < 0.05 ) with increased oxidative stress . However , simvastatin inhibited this promotion ( 2.5+/-0.3 mm , p < 0.001 vs. P01375 alone ) by decreasing oxidative stress , P15692 , Akt , and P29474 . SIGNIFICANCE : We conclude that at the same dosage , simvastatin can either promote or inhibit angiogenesis , possibly by activating upstream regulators of HIF-1alpha in hypoxia , but conversely interfering with angiogenic signaling downstream to inflammation . These opposing angiogenic effects should be considered in the therapeutic strategies with statins . Deficiency of the oxygen sensor Q96KS0 augments liver regeneration after partial hepatectomy . PURPOSE : Liver regeneration after partial hepatectomy ( PH ) occurs in conditions of reduced oxygen supply . HIF prolyl hydroxylase enzymes ( Q96KS0 , Q9GZT9 , and Q9H6Z9 ) are oxygen sensors involved in adaptive response to hypoxia . Specific functions of these P20941 enzymes in liver regeneration have , however , remained enigmatic . Here , we investigated the significance of Q96KS0 in liver regeneration following hepatectomy . METHODS : Liver regeneration was studied in Q96KS0 -deficient ( Q96KS0 (-/-) ) and wild type ( WT ) mice subjected to 80 % hepatectomy . For in vitro analyses , hepatocytes were isolated from Q96KS0 (-/-) and WT livers . Cell cycle progression was studied via FACS-based analysis of nuclear DNA profile . Transcription factor binding assays , qRT-PCR , and immunoblotting were applied to study the relevance of Q96KS0 downstream effectors during liver regeneration . RESULTS : Liver regeneration was significantly enhanced in Q96KS0 (-/-) mice compared to WT littermates . This effect was due to enhanced proliferation rather than to hypertrophy of liver cells . Cell cycle progression was significantly enhanced , and transcriptional activity of the cell cycle regulator c-Myc was increased in Q96KS0 -deficient hepatocytes . These changes coincided with increased expression of cyclin D2 , a cell cycle-promoting c-Myc target , and decreased expression of the cell cycle-delaying c-Myc target P38936 . CONCLUSIONS : Loss of Q96KS0 enhances liver regeneration by boosting hepatocyte proliferation in a c-Myc-dependent fashion . Q96KS0 might , therefore , represent a potential target to facilitate liver regeneration after surgical resection . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Nitrergic response to cyclophosphamide treatment in blood and bone marrow . Daily intraperitoneal injection of cyclophosphamide ( P15085 ) ( 50 mgkg(-1) of body weight ) for 5 days resulted in reduced levels of marrow and blood cellularity , which was most pronounced in 18 days post-treatment ( pt ) . On day 18 after P15085 treatment the enhancedlevels of nitric oxide ( NO ) precursors and metabolites ( L-arginine , L-citrulline , reactive nitrogen species ( RNS ) ) of marrow and blood cells ( platelet , neutrophil , lymphocyte and monocyte ) resulted from up-regulation of Ca(II)/calmodulin( P62158 )-independent " inducible " NO synthase ( P35228 ) , with a lessercontribution of Ca(II)/ P62158 -dependent " constitutive " P29474 isoforms to systemic NO.Biphasic response to P15085 of marrow nitrergic system , i.e. both P35228 and P29474 showed significantly depressed activities , as well as diminished levels of NO metabolites on day 9 pt , suggested that signals in addition to NO might be involved in P15085 -induced inhibition of hematopoesis , while a gradual increase of neutrophil and platelet NOS activity appeared to be contributed to a P15085 -induced development of granulopenia , thrombocytopenia and hemorrhage . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . DB00435 donor , (+/-)-S-nitroso-N-acetylpenicillamine , stabilizes transactive hypoxia-inducible factor-1alpha by inhibiting von Hippel-Lindau recruitment and asparagine hydroxylation . We have confirmed that the NO donor (+/-)-S-nitroso-N-acetylpenicillamine ( P60880 ) stabilizes the transactive form of hypoxia-inducible factor-1alpha ( HIF-1alpha ) , leading to the induction of HIF-1alpha target genes such as vascular endothelial growth factor and carbonic anhydrase 9 . Activation of HIF-1alpha should require inhibition of the dual system that keeps it inactive . One is ubiquitination , which is triggered by hydroxylation of HIF-1alpha-proline and the subsequent binding of E3 ubiquitin ligase , the von Hippel Lindau ( P40337 ) protein . The other is hydroxylation of HIF-1alpha-asparagine , which reduces the affinity of HIF-1alpha for its coactivator , DB02527 responsive element binding protein/p300 . We examined the effects of the NO donor P60880 on proline and asparagine hydroxylation of HIF-1alpha peptides by measuring the activities of the corresponding enzymes , HIF-1alpha-specific proline hydroxylase 2 ( Q9GZT9 ) and the HIF-1alpha-specific asparagine hydroxylase , designated factor inhibiting HIF-1alpha ( Q9NWT6 ) , respectively . We found that the P60880 did not prevent Q9GZT9 from hydroxylating the proline of HIF-1alpha . Instead , it blocked the interaction between P40337 and the proline-hydroxylated HIF-1alpha , but only when the reducing agents Fe(II) and vitamin C were limiting . The fact that the absence of cysteine 520 of HIF-1alpha abolishes its responsiveness to P60880 suggests that this residue mediates the inhibition by P60880 of the interaction between P40337 and HIF-1alpha , presumably by S-nitrosylation of HIF-1alpha . Un-like Q9GZT9 , asparagine hydroxylation by Q9NWT6 was directly inhibited by P60880 , but again only when reducing agents were limiting . Substitution of cysteine 800 of HIF-1alpha with alanine failed to reverse the inhibitory effects of P60880 on asparagine hydroxylation , implying that Q9NWT6 , not its substrate HIF-1alpha , is inhibited by P60880 . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . DB00126 is dispensable for oxygen sensing in vivo . Prolyl-4-hydroxylation is necessary for proper structural assembly of collagens and oxygen-dependent protein stability of hypoxia-inducible transcription factors ( HIFs ) . In vitro function of HIF prolyl-4-hydroxylase domain ( P20941 ) enzymes requires oxygen and 2-oxoglutarate as cosubstrates with iron(II) and vitamin C serving as cofactors . Although vitamin C deficiency is known to cause the collagen-disassembly disease scurvy , it is unclear whether cellular oxygen sensing is similarly affected . Here , we report that vitamin C-deprived Gulo(-/-) knockout mice show normal HIF-dependent gene expression . The systemic response of Gulo(-/-) animals to inspiratory hypoxia , as measured by plasma erythropoietin levels , was similar to that of animals supplemented with vitamin C . Hypoxic HIF induction was also essentially normal under serum- and vitamin C-free cell-culture conditions , suggesting that vitamin C is not required for oxygen sensing in vivo . Glutathione was found to fully substitute for vitamin C requirement of all 3 P20941 isoforms in vitro . Consistently , glutathione also reduced HIF-1α protein levels , transactivation activity , and endogenous target gene expression in cells exposed to CoCl(2) . A Cys201Ser mutation in Q9GZT9 increased basal hydroxylation rates and conferred resistance to oxidative damage in vitro , suggesting that this surface-accessible Q9GZT9 cysteine residue is a target of antioxidative protection by vitamin C and glutathione . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . Enhanced sensitivity to irinotecan by Cdk1 inhibition in the p53-deficient HT29 human colon cancer cell line . Mutations in the tumor-suppressor gene p53 have been associated with advanced colorectal cancer ( CRC ) . DB00762 ( CPT-11 ) , a P11387 inhibitor , has been recently incorporated to the adjuvant therapy . Since the DNA-damage checkpoint depends on p53 activation , the status of p53 might critically influence the response to CPT-11 . We analysed the sensitivity to CPT-11 in the human colon cancer cell line HT29 ( mut p53 ) and its wild-type (wt)-p53 stably transfected subclone HT29-A4 . Cell-cycle analysis in synchronised cells demonstrated the activation of transfected wt-p53 and a P38936 ( P38936 /CIP1)-dependent cell-cycle blockage in the S phase . Activated wt-p53 increased apoptosis and enhanced sensitivity to CPT-11 . In p53-deficient cells , cDNA-macroarray analysis and western blotting showed an accumulation of the cyclin-dependent kinase (cdk)1/cyclin B complex . Subsequent p53-independent activation of the cdk-inhibitor ( cdk-I ) P38936 ( P38936 /CIP1) prevented cell-cycle progression . Cdk1 induction was exploited in vivo to improve the sensitivity to CPT-11 by additional treatment with the cdk-I P99999 -202 . We demonstrate a gain of sensitivity to CPT-11 in a p53-mutated colon cancer model either by restoring wild-type p53 function or by sequential treatment with cdk-Is . Considering that mutations in p53 are among the most common genetic alterations in CRC , a therapeutic approach specifically targeting p53-deficient tumors could greatly improve the treatment outcomes . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Candidate gene studies of ADHD : a meta-analytic review . Quantitative genetic studies ( i.e. , twin and adoption studies ) suggest that genetic influences contribute substantially to the development of attention deficit hyperactivity disorder ( ADHD ) . Over the past 15 years , considerable efforts have been made to identify genes involved in the etiology of this disorder resulting in a large and often conflicting literature of candidate gene associations for ADHD . The first aim of the present study was to conduct a comprehensive meta-analytic review of this literature to determine which candidate genes show consistent evidence of association with childhood ADHD across studies . The second aim was to test for heterogeneity across studies in the effect sizes for each candidate gene as its presence might suggest moderating variables that could explain inconsistent results . Significant associations were identified for several candidate genes including Q01959 , P21917 , P21918 , P31645 , P28222 , and P60880 . Further , significant heterogeneity was observed for the associations between ADHD and Q01959 , P21917 , P21918 , P09172 , P08913 , P31645 , Q8IWU9 , P21397 , and P60880 , suggesting that future studies should explore potential moderators of these associations ( e.g. , ADHD subtype diagnoses , gender , exposure to environmental risk factors ) . We conclude with a discussion of these findings in relation to emerging themes relevant to future studies of the genetics of ADHD .

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interacts_with DB00624?

multiple_choice

P10275 -immunoreactive cells in ram hypothalamus : distribution and co-localization patterns with gonadotropin-releasing hormone , somatostatin and tyrosine hydroxylase . DB00624 exerts important feedback effects on the hypothalamus of the ram to influence reproductive functioning . To provide a neuroanatomical basis for understanding this androgen action , the present study has examined androgen receptor ( AR ) immunoreactivity within the hypothalamus and adjacent brain areas of the intact non-breeding season ram . The largest populations of AR-immunoreactive cells were detected in the medial preoptic area , infundibular and premammillary nuclei in addition to the ventromedial nucleus ( VMN ) where cells were found distributed throughout its medial and lateral divisions . Smaller numbers of AR-expressing cells were identified in the bed nucleus of the stria terminalis and anterior hypothalamic area ( DB00551 ) including the paraventricular , but not the supraoptic , nucleus . Double-labelling immunocytochemistry revealed the presence of AR immunoreactivity in only 2 of 460 gonadotropin-releasing hormone ( DB00644 ) neurons . A very small population of TH-immunoreactive cells located in the lateral aspect of the DB00551 was found to contain ARs . Dopaminergic cells elsewhere in the hypothalamus , including the infundibular nucleus , did not display AR immunoreactivity . Nearly 50 % of AR-expressing cells in the lateral VMN were immunoreactive for somatostatin while less than 5 % of periventricular somatostatin neurons displayed AR immunoreactivity . These results show where ARs are expressed in the ram hypothalamus and indicate the neuroanatomical sites at which androgen may act to influence reproductive function . The absence of ARs in the neuroendocrine DB00644 and tuberoinfundibular dopaminergic cells suggests that androgens do not influence the genome of these cells in any direct manner . In contrast , the somatostatin neurons of the VMN appear to be an important target for circulating androgens in the non-breeding season ram . Mediator subunit Gal11p/ Q96RN5 is required for fatty acid-dependent gene activation by yeast transcription factor Oaf1p . The yeast zinc cluster transcription factor Oaf1p activates transcription of target genes in response to direct binding of fatty acids in a manner analogous to the vertebrate nuclear receptor peroxisome proliferator-activated receptoralpha ( PPARalpha ) . PPARs and other metazoan nuclear receptors productively engage several distinct LXXLL motif-containing co-activators , including P52701 family members and the Q15648 /MED1 subunit of the Mediator co-activator , to promote ligand-dependent gene activation . Yeast , however , does not appear to harbor LXXLL motif co-activators , and the mechanism of fatty acid-dependent gene activation by the yeast PPARalpha analog Oaf1p is unknown . Here we show that the yeast Mediator subunit Gal11p/ Q96RN5 and its activator-targeted KIX domain plays a critical role in fatty acid-dependent transcriptional regulation of fatty acid beta-oxidation and peroxisomal genes by Oaf1p and for the ability of yeast to utilize fatty acids as a sole carbon source . Moreover , structural studies by NMR spectroscopy reveal that the Oaf1p activation domain interacts with the Gal11p/ Q96RN5 KIX domain in a manner similar to the yeast zinc cluster family member and xenobiotic receptor Pdr1p , revealing that the Gal11p/ Q96RN5 KIX domain is a key target of several ligand-dependent transcription factors in yeast . Together with previous work showing that the Caenorhabditis elegans Gal11p/ Q96RN5 homolog MDT-15 plays a critical role in regulation of fatty acid metabolism by the nematode Q07869 -like nuclear receptor NHR-49 , the findings presented here provide evidence for an ancient and essential role of a Mediator co-activator subunit in regulation of fatty acid metabolism by nuclear receptor-like transcription factors in eukaryotes . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . P10275 -dependent transactivation of growth arrest-specific gene 6 mediates inhibitory effects of testosterone on vascular calcification . Recent epidemiological studies have found that androgen deficiency is associated with a higher incidence of cardiovascular disease in men . However , little is known about the mechanism underlying the cardioprotective effects of androgens . Here we show the inhibitory effects of testosterone on vascular calcification and a critical role of androgen receptor ( AR ) -dependent transactivation of growth arrest-specific gene 6 ( Gas6 ) , a key regulator of inorganic phosphate ( P(i) ) -induced calcification of vascular smooth muscle cells ( VSMC ) . DB00624 and nonaromatizable androgen dihydrotestosterone inhibited P(i)-induced calcification of human aortic VSMC in a concentration-dependent manner . Androgen inhibited P(i)-induced VSMC apoptosis , an essential process for VSMC calcification . The effects on VSMC calcification were mediated by restoration of P(i)-induced down-regulation of Gas6 expression and a subsequent reduction of Akt phosphorylation . These effects of androgen were blocked by an AR antagonist , flutamide , but not by an estrogen receptor antagonist , DB00947 . We then explored the mechanistic role of the AR in Gas6 expression and found an abundant expression of AR predominantly in the nucleus of VSMC and two consensus ARE sequences in the Gas6 promoter region . DB02901 stimulated Gas6 promoter activity , and this effect was abrogated by flutamide and by AR siRNA . Site-specific mutation revealed that the proximal ARE was essential for androgen-dependent transactivation of Gas6 . Furthermore , chromatin immunoprecipitation assays demonstrated ligand-dependent binding of the AR to the proximal ARE of Gas6 . These results indicate that AR signaling directly regulates Gas6 transcription , which leads to inhibition of vascular calcification , and provides a mechanistic insight into the cardioprotective action of androgens . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Novel function of androgen receptor-associated protein 55/ O43294 as a negative regulator of P84022 signaling . P10275 -associated protein 55 ( O43294 / O43294 ) belongs to the LIM protein superfamily and is featured by three or four N-terminal LD motifs and four C-terminal zinc finger-like LIM domains . Both LD motifs and LIM domains can serve as protein-protein interaction interfaces . Recently , we found that enforced expression of O43294 inhibits transforming growth factor-beta-mediated up-regulation of Smad binding element-luciferase reporter activity in NRP-154 and NRP-152 rat prostate and LNCaP human prostate cell lines . Moreover , O43294 also inhibits the induction of Smad-binding element 4-luciferase and 3TP-luciferase ( a plasminogen activator inhibitor-1 ( P05121 ) promoter construct ) reporters by constitutively active ( CA ) - P84022 in these cell lines . Co-immunoprecipitation studies suggest an interaction between O43294 and either CA- P84022 or wild-type P84022 in HEK293 cells that occurs through the MH2 domain of P84022 and the C terminus of O43294 with wild-type P84022 having stronger affinity than CA- P84022 to O43294 . O60760 pull-down assays demonstrate that this interaction can occur in a cell-free system . These results are consistent with the luciferase data showing that the C terminus of O43294 is critical for suppression of P84022 activity . Furthermore , using a mammalian two-hybrid system , we confirmed that O43294 interacts with the MH2 domain of P84022 and suppresses CA- P84022 -induced transcriptional responses . In conclusion , these results support that O43294 selectively intercepts transforming growth factor-beta signaling through an interaction of the LIM domain of O43294 with the MH2 domain of P84022 . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . P10275 is widely expressed in bovine placentomes and up-regulated during differentiation of bovine trophoblast giant cells . OBJECTIVES : In cattle no biological role has been definitely identified for placental estrogens and progesterone . However , in the bovine trophoblast androgens may also be produced and have local effects . Thus , the aims of this study were to identify androgen receptor ( AR ) expressing cells and to monitor testosterone tissue concentrations in bovine placentomes throughout gestation . METHODS : Placental AR expression was characterized at the mRNA and protein level applying conventional and real-time RT-qPCR , western blot and immunohistochemistry . DB00624 was measured by radioimmunoassay . RESULTS : AR-mRNA was qualitatively detected from day 50 of gestation until term . Mean relative gene expression levels were constant between day 100 and late gestation . A slight non-significant increase was observed in the prepartal period . With immunohistochemistry distinct nuclear signals were predominantly observed in invasive trophoblast giant cells ( P21980 ) from day 80 until term . In mature P21980 of the trophoblast , immature P21980 and uninucleated trophoblast cells , stromal cells of the chorionic villi , caruncular epithelial and stromal cells immunoreactive score values were low at early and midgestation but increased significantly ( p < 0.01 ) during late gestation and remained high until parturition . With western blot in placentomal tissue a specific band of approximately 110 kDa was detected as it was the case in epididymis used as a positive control . DB00624 concentrations increased from 0.70 ± 0.29 pmol/g wet tissue between days 60-220 to 4.22 ± 1.29 pmol/g during late gestation ( p < 0.001 ) . DISCUSSION : The results are consistent with androgens as active products of bovine placental steroidogenesis . The substantial up-regulation of AR expression during P21980 differentiation suggests that androgens may be related to this process . P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . P10275 -dependent activation of endothelial nitric oxide synthase in vascular endothelial cells : role of phosphatidylinositol 3-kinase/akt pathway . The mechanisms of testosterone-induced vasodilatation are not fully understood . This study investigated the effect of testosterone on nitric oxide ( NO ) synthesis and its molecular mechanism using human aortic endothelial cells ( HAEC ) . DB00624 at physiological concentrations ( 1-100 nm ) induced a rapid ( 15-30 min ) increase in NO production , which was associated with phosphorylation and activation of endothelial NO synthase ( P29474 ) . Then , the involvement of the androgen receptor ( AR ) , which is abundantly expressed in HAEC , was examined . The effect of testosterone on P29474 activation and NO production were abolished by pretreatment with an AR antagonist nilutamide and by transfection with AR small interference RNA . In contrast , testosterone-induced P29474 phosphorylation was unchanged by pretreatment with an aromatase inhibitor or by transfection with ERalpha small interference RNA . DB02901 , a nonaromatizable androgen , also stimulated P29474 phosphorylation . Next , the signaling cascade that leads to P29474 phosphorylation was explored . DB00624 stimulated rapid phosphorylation of Akt in a time- and dose-dependent manner , with maximal response at 15-60 min . The rapid phosphorylation of P29474 or NO production induced by testosterone was inhibited by Akt inhibitor SH-5 or by phosphatidylinositol ( PI ) 3-kinase inhibitor wortmannin . Co-immunoprecipitation assays revealed a testosterone-dependent interaction between AR and the p85alpha subunit of P19957 -kinase . In conclusion , testosterone rapidly induces NO production via AR-dependent activation of P29474 in HAEC . Activation of P19957 -kinase/Akt signaling and the direct interaction of AR with p85alpha are involved , at least in part , in P29474 phosphorylation . Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway . DB00624 supplementation increases skeletal muscle mass and decreases fat mass ; however , the underlying mechanisms are unknown . We hypothesized that testosterone regulates body composition by promoting the commitment of mesenchymal pluripotent cells into myogenic lineage and inhibiting their differentiation into adipogenic lineage . Mouse C3H 10T1/2 pluripotent cells were treated with testosterone ( 0-300 nM ) or dihydrotestosterone ( DB02901 , 0-30 nM ) for 0-14 d , and myogenic conversion was evaluated by immunocytochemical staining for early ( MyoD ) and late ( myosin heavy chain II ; MHC ) myogenic markers and by measurements of MyoD and MHC mRNA and protein . Adipogenic differentiation was assessed by adipocyte counting and by measurements of peroxisomal proliferator-activated receptor gamma 2 ( Q07869 gamma 2 ) mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . The number of MyoD+ myogenic cells and MHC+ myotubes and MyoD and MHC mRNA and protein levels increased dose dependently in response to testosterone and DB02901 treatment . Both testosterone and DB02901 decreased the number of adipocytes and down-regulated the expression of Q07869 gamma 2 mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . P10275 mRNA and protein levels were low at baseline but increased after testosterone or DB02901 treatment . The effects of testosterone and DB02901 on myogenesis and adipogenesis were blocked by bicalutamide . Therefore , testosterone and DB02901 regulate lineage determination in mesenchymal pluripotent cells by promoting their commitment to the myogenic lineage and inhibiting their differentiation into the adipogenic lineage through an androgen receptor-mediated pathway . The observation that differentiation of pluripotent cells is androgen dependent provides a unifying explanation for the reciprocal effects of androgens on muscle and fat mass in men . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Chemical ablation of androgen receptor in prostate cancer cells by the histone deacetylase inhibitor LAQ824 . P10275 plays a critical role in the development of primary as well as advanced hormone-refractory prostate cancer . Therefore , ablation of androgen receptor from prostate cancer cells is an interesting concept for developing a new therapy not only for androgen-dependent prostate cancer but also for metastatic hormone-refractory prostate cancer , for which there is no effective treatment available . We report here that LAQ824 , a cinnamyl hydroxamatic acid histone deacetylase inhibitor currently in human clinical trials , effectively depleted androgen receptor in prostate cancer cells at nanomolar concentrations . LAQ824 seemed capable of depleting both the mutant and wild-type androgen receptors in either androgen-dependent and androgen-independent prostate cancer cells . Although LAQ824 may exert its effect through multiple mechanisms , several lines of evidence suggest that inactivation of the heat shock protein-90 ( Hsp90 ) molecular chaperone is involved in LAQ824-induced androgen receptor depletion . Besides androgen receptor , LAQ824 reduced the level of Hsp90 client proteins HER-2 ( ErbB2 ) , Akt/ P31749 , and P04049 in LNCaP cells . Another Hsp90 inhibitor , 17-allyamino-17-demethoxygeldanamycin ( 17- P29372 ) , also induced androgen receptor diminution . LAQ824 induced Hsp90 acetylation in LNCaP cells , which resulted in inhibition of its DB00171 -binding activity , dissociation of Hsp90-androgen receptor complex , and proteasome-mediated degradation of androgen receptor . Consequently , LAQ824 blocked androgen-induced prostate-specific antigen production in LNCaP cells . LAQ824 effectively inhibited cell proliferation and induced apoptosis of these prostate cancer cells . These results reveal that LAQ824 is a potent agent for depletion of androgen receptor and a potential new drug for prostate cancer . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Increased dihydrotestosterone receptor levels in high-stage renal adenocarcinoma . Primary renal adenocarcinoma tissue , metastatic deposits , and normal kidney parenchyma from 16 patients were assayed for sex hormone receptors by dextran-coated charcoal adsorption and sucrose gradient centrifugation techniques . DB02901 receptors ( P10275 ) were found in all renal carcinomatous tissue ( 20/20 ) and in 93 % ( 13/14 ) autologous normal kidneys analyzed . DB00624 receptors were found in 84 % ( 16/19 ) of tumors and 93 % ( 14/15 ) or normal kidneys analyzed . Estrogen receptors in small amounts ( ER ) were detected in only 5 % ( 1/19 ) of tumors and in 7 % ( 1/15 ) of normal kidneys . Progesterone receptors ( PR ) in low quantities were detected in 30 % ( 6/20 ) of renal tumors and in 40 % ( 6/15 ) of normal kidneys . P10275 levels in high-stage tumors ( DB00279 , DB00451 ) were significantly elevated over levels in autologous normal kidney , whereas in low-stage tumors localized to the kidney ( T1 and P24752 tumors ) P10275 levels were not significantly different from autologous normal kidney . The mean levels of P10275 in high-stage kidney tumors were significantly elevated over levels in low-stage tumors ( P less than 0.001 ) . P10275 estimation in renal neoplasms may help in biologic staging of renal adenocarcinoma and could define a group of patients in whom anti-androgen therapy may be worth a trial . Mass spectrometry and hydrogen/deuterium exchange measurements of alcohol-induced structural changes in cellular retinol-binding protein type I . To bind and release its ligand , cellular retinol-binding protein type I ( P09455 ) needs to undergo conformational and dynamic changes to connect the inner , solvent-shielded cavity , where retinol is found to bind , and the outside medium . DB00162 dissociation in vitro is favoured by water/alcohol mixtures whose moderately low dielectric constants mimic a property characteristic of the membrane microenvironment where this process occurs in vivo . Apo- and holo- P09455 , in either water/methanol or water/trifluoroethanol ( TFE ) mixtures , were analyzed at equilibrium by electrospray ionization with orthogonal quadrupole time-of-flight mass spectrometry ( P19957 -Q-TOFMS ) to identify the alcohol-induced species . The questions were asked whether the presence of alcohols affects protein dynamics , as reflected by hydrogen/deuterium ( H/D ) exchange monitored by continuous-labelling experiments , and to which extent retinol dissociation influences the process . With increasing methanol , at pH near neutrality , apo- P09455 exhibits a progressively more compact conformation , resulting in reduced H/D exchange with respect to the native protein in water . DB00162 dissociation from the holo-protein did not promote hydrogen replacement . Similarly , in the presence of the low TFE concentration sufficient to cause retinol dissociation , the hydrogen exchange of the resulting apo-protein was not exalted . However , in contrast with the alkanol , higher TFE concentrations induced a transition of apo- P09455 to a new alpha-helix conformation capable of exchanging all available hydrogen atoms . Transforming growth factor-beta1 induces tumor stroma and reduces tumor infiltrate in cervical cancer . Cervical carcinomas consist of tumor cell nests surrounded by varying amounts of intratumoral stroma containing different quantities and types of immune cells . Besides controlling ( epithelial ) cell growth , the multifunctional cytokine transforming growth factor-beta(1) ( TGF-beta(1) ) is involved in the formation of stroma and extracellular matrix ( Q13201 ) and in immunosuppression . Several malignancies are known to be associated with enhanced production of TGF-beta(1) , repression or mutation of TGF-beta transmembrane receptors , or mutations at the postreceptor intracellular signaling pathway . The aim of our study was to investigate the role of tumor cell-derived TGF-beta(1) on the amount of intratumoral stroma ; the deposition of collagen IV , fibronectin , and laminin ; and the tumor infiltrate in cervical carcinoma . The expression of TGF-beta(1) mRNA in 108 paraffin-embedded cervical carcinomas was detected by mRNA in situ hybridization . Immunohistochemistry was used to investigate the amount of tumor stroma and Q13201 proteins and the extent of the tumor infiltrate . P00747 activator inhibitor-1 ( P05121 ) protein expression in tumor cells was determined to verify the biological activity of TGF-beta(1.) Cytoplasmatic TGF-beta(1) mRNA expression in tumor cells was significantly correlated with the amount of intratumoral stroma and the deposition of collagen IV . TGF-beta(1) mRNA expression in every tumor was accompanied by P05121 expression , indicating biological activity of TGF-beta(1) . An inverse relationship between TGF-beta(1) mRNA expression in tumor cells and the extent of the tumor infiltrate was demonstrated . Our results indicate that cervical cancer cells affect the amount and the composition of the intratumoral stroma and the tumor infiltrate by the production and secretion of TGF-beta(1) . P10275 in human placental villi . In studies with a synthetic androgen , R 1881 , an androgen-binding component was found in the cytosol of human placental villi . Kinetic analysis indicated that the Kd value of this component was 1.4 nM at 0-4 degrees C and that binding of R 1881 amounted to 277 +/- 73 fmol/mg protein. glycerol density gradient ultracentrifugation showed a peak of binding activity in the 8S region in a medium of low ionic strength , but in the 4.5S region in a medium containing 9.5 M DB00761 . The R 1881-binding component was inactivated by mild heat- or trypsin-treatment , but not by treatment with DNase or RNase . Most of the R 1881-binding activity was sedimented at 20 to 40 % saturation of ammonium sulfate . These findings indicate that the R 1881-binding component in human placental cytosol is quite similar in its characteristics to androgen receptors , which are present in various androgen-responsive organs . DB00624 was a more potent competitor of R 1881-binding than DB02901 or cyproterone acetate . Scatchard plots indicated that the binding site of testosterone was identical with that of R 1881 . These findings suggest that the androgen receptor in placental cytosol is specific for testosterone . The Kd value for testosterone was calculated to be 3.2 nM . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . The androgen axis in recurrent prostate cancer . PURPOSE : Prostate cancer that recurs during androgen deprivation therapy is referred to as androgen-independent . High levels of expression of androgen receptor and androgen receptor-regulated genes in recurrent prostate cancer suggest a role for androgen receptor and its ligands in prostate cancer recurrence . EXPERIMENTAL DESIGN : Recurrent prostate cancer specimens from 22 men whose prostate cancer recurred locally during androgen deprivation therapy and benign prostate specimens from 48 men who had received no prior treatment were studied . P10275 expression was measured using monoclonal antibody and automated digital video image analysis . Tissue androgens were measured using radioimmunoassay . RESULTS : Epithelial nuclei androgen receptor immunostaining in recurrent prostate cancer ( mean optical density , 0.284 +/- SD 0.115 and percentage positive nuclei , 83.7 +/- 11.6 ) was similar to benign prostate ( mean optical density , 0.315 +/- 0.044 and percentage positive nuclei , 77.3 +/- 13.0 ) . Tissue levels of testosterone were similar in recurrent prostate cancer ( 2.78 +/- 2.34 pmol/g tissue ) and benign prostate ( 3.26 +/- 2.66 pmol/g tissue ) . Tissue levels of dihydrotestosterone , dehydroepiandrosterone , and androstenedione were lower ( Wilcoxon , P = 0.0000068 , 0.00093 , and 0.0089 , respectively ) in recurrent prostate cancer than in benign prostate , and mean dihydrotestosterone levels , although reduced , remained 1.45 nM . P10275 activation in recurrent prostate cancer was suggested by the androgen-regulated gene product , prostate-specific antigen , at 8.80 +/- 10.80 nmol/g tissue . CONCLUSIONS : DB00624 and dihydrotestosterone occur in recurrent prostate cancer tissue at levels sufficient to activate androgen receptor . Novel therapies for recurrent prostate cancer should target androgen receptor directly and prevent the formation of androgens within prostate cancer tissue . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions .

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DB08877 for the treatment of primary myelofibrosis . PURPOSE : The pharmacology , pharmacokinetics , pharmacogenomics , clinical efficacy , and safety profile of ruxolitinib for the treatment of primary myelofibrosis are reviewed . SUMMARY : DB08877 , an oral tyrosine kinase inhibitor that targets the Janus-associated kinases ( JAKs ) 1 and 2 , has been recently approved for the treatment of patients with intermediate- or high-risk myelofibrosis . Unlike previous treatment options for patients with myelofibrosis , ruxolitinib offers a targeted therapy option for these patients who often suffer with severe and debilitating symptoms associated with the disease process . After oral administration , ruxolitinib is rapidly absorbed and can be given without regard to meals . DB08877 is primarily metabolized by the cytochrome P-450 ( CYP ) 3A4 isoenzyme system ; therefore , if concomitant use with a strong P08684 inhibitor is unavoidable , an initial dosage reduction is warranted . Two Phase III randomized trials comparing ruxolitinib to either placebo or best available therapy found a rapid and sustained response in the reduction of spleen size and improvements in constitutional symptoms and quality of life , with one study demonstrating an improvement in overall survival . The most commonly reported serious adverse effects of ruxolitinib are anemia and thrombocytopenia . DB08877 is administered as an oral tablet given twice daily , with the initial starting dosage based on the baseline platelet count . Dosage reductions are based on the development of thrombocytopenia . CONCLUSION : By directly targeting both P23458 and O60674 through small-molecule inhibition , ruxolitinib elicits a reduction in splenomegaly and disease-related symptoms in patients with intermediate- or high-risk myelofibrosis while maintaining an acceptable toxicity profile and a low treatment-discontinuation rate . P00533 -induced cell migration is mediated predominantly by the JAK- P35610 pathway in primary esophageal keratinocytes . The epidermal growth factor receptor ( P00533 ) activates several signaling cascades in response to epidermal growth factor stimulation . One of these signaling events involves tyrosine phosphorylation of signal transducer and activator of transcription ( P35610 ) , whereas another involves activation of the phosphatidylinositol 3-OH kinase pathway . Two possibilities for P35610 activation exist : a janus kinase ( JAK ) -dependent and a JAK-independent mechanism . Herein , we demonstrate that P00533 overexpression in primary esophageal keratinocytes activates P35610 in a JAK-dependent fashion with the functional consequence of enhanced cell migration , which can be abolished by use of a JAK-specific inhibitor , AG-490 . We determined the mechanisms underlying the signal transduction pathway responsible for increased cell migration . Stimulation of P00533 induces Tyr701 phosphorylation of P42224 and initiates complex formation of P42224 and P40763 with P23458 and O60674 . Thereafter , the STATs translocate to the nucleus within 15 min . In addition , we found that activation of this signaling pathway results in matrix metalloproteinase-1 ( P03956 ) activity . By contrast , Akt activation does not impact the P00533 -STATs-JAKs complex formation and nuclear translocation of the STATs with subsequent P03956 activity , although Akt activation may contribute to cell migration through an independent mechanism . Taken together , we find that the recruitment of the P35610 -JAK complex by P00533 is responsible for keratinocyte migration that , in turn , might be mediated by P03956 activation . Phase II study of erlotinib ( DB00530 ) in patients with metastatic colorectal cancer . Erlotinib ( Tarceva , DB00530 ) , a potent epidermal growth factor receptor tyrosine kinase inhibitor ( P00533 ) , was evaluated in a phase II study to assess its activity in patients with metastatic colorectal cancer . In all , 38 patients with metastatic colorectal cancer were treated with erlotinib at a continuous daily oral dose of 150 mg . Radiological evaluation was carried out every 8 weeks and tumour biopsies were performed before treatment and on day 8 . Of 31 evaluable patients , 19 ( 61 % ) had progressive disease and 12 ( 39 % ) had stable disease ( s.d. ) . The median time to progression for those patients having s.d. was 123 days ( range 108-329 days ) . The most common adverse events were rash in 34 patients and diarrhoea in 23 patients . Correlative studies were conducted to investigate the effect of erlotinib on downstream signalling . Tumour tissue correlations were based on usable tissue from eight match paired tumour samples pre- and on therapy , and showed a statistically significant decrease in the median intensity of both pEGFR ( P=0.008 ) and phospho-extracellular signal-regulated kinase ( P29323 ) ( P=0.008 ) a week after commencement of treatment . No other statistically significant change in tumour markers was observed . Erlotinib was well tolerated with the most common toxicities being rash and diarrhoea . More than one-third of evaluable patients had s.d. for a minimum of 8 weeks . Correlative studies showed a reduction in phosphorylated P00533 and P29323 in tumour tissue post-treatment . Interstitial lung disease in patients with non-small-cell lung cancer treated with epidermal growth factor receptor inhibitors . Interstitial lung disease ( ILD ) refers to a diverse range of pulmonary fibrotic disorders and may be hard to accurately diagnose , as distinguishing it from other pulmonary diseases can be difficult . Estimations of the incidence in populations are confounded by the complexity of the different forms of the disorder . In addition , ILD is a comorbid disease of lung cancer and is seen after most forms of chemotherapy and radiotherapy for advanced lung cancer . Incidences of > or=10 % have been reported ; however , whatever the true incidence , both chemotherapy and radiotherapy enhance the risk of developing ILD . ILD has also been reported with the epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitors , including erlotinib ( Tarceva , DB00530 ) and gefitinib ( IRESSA ) . In a large number of gefitinib-treated patients ( n > 185,000 ) an incidence of approx 1 % has been observed ( approx 2 % in Japan ; 0.3 % in the rest of the world ) . Nevertheless , as with other treatments for advanced non-small-cell lung cancer , the clinical benefit outweighs the risk of ILD . In this article , we review the data on ILD with P00533 inhibitors and other common lung cancer treatments . Overview of tyrosine kinase inhibitors in clinical breast cancer . Studies of cell models and profiling of clinical breast cancer material to reveal the mechanisms of resistance to anti-oestrogen therapy , and to tamoxifen in particular , have reported that this phenomenon can be associated with increased expression and signalling through erbB Type 1 growth factor receptors , notably the epidermal growth factor receptor ( P00533 ) and P04626 . Further molecular studies have revealed an intricate interlinking between such growth factor receptor pathways and oestrogen receptor ( ER ) signalling . Inhibition of receptor tyrosine kinase activity involved in the P00533 signalling cascade forms the basis for the use of P00533 specific tyrosine kinase inhibitors exemplified by gefitinib ( ZD1839 , DB00317 ) and erlotinib ( DB00530 , Tarceva ) . Such agents have proved promising in pre-clinical studies and are currently in clinical trials in breast cancer , where gefitinib has been studied more extensively to date . Here , we present an overview of the current development of gefitinib in clinical breast cancer . This includes results from our clinical breast cancer trial 1839IL/0057 that demonstrate the efficacy of gefitinib within ER-positive , tamoxifen-resistant patients with locally advanced/metastatic disease , where parallel decreases in P00533 signal transduction and the Ki67 ( Q86YT6 ) proliferation marker can be detected as predicted from model system studies . We also consider trials examining combination treatment with gefitinib and anti-hormonal strategies that will begin to address the clinically important question of whether gefitinib can delay/prevent onset of anti-hormone resistance . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . Efficacy and safety of erlotinib HCl , an epidermal growth factor receptor ( P00533 / P00533 ) tyrosine kinase inhibitor , in patients with advanced ovarian carcinoma : results from a phase II multicenter study . The aim of this single-arm , phase II study was to estimate the tumor response rate and safety profile of erlotinib HCl ( erlotinib , Tarceva , DB00530 ) monotherapy in patients with refractory , recurrent , P00533 / P00533 -positive epithelial ovarian tumors , who had failed prior taxane and/or platinum-based chemotherapy . Thirty-four patients received 150 mg erlotinib orally once daily for up to 48 weeks or until disease progression or dose-limiting toxicity . Two patients had partial responses , lasting 8+ and 17 weeks , giving an objective response rate of 6 % ( 95 % confidence interval [ CI ] , 0.7-19.7 % ) . Fifteen patients ( 44 % ) had stable disease , and 17 patients ( 50 % ) had progressive disease . Median overall survival was 8 months ( 95 % CI , 5.7-12.7 months ) , with a 1-year survival rate of 35.3 % ( 95 % CI , 19.8-53.5 % ) . Patients with rash survived significantly longer than those without ( P= 0.009 ) , correlating with rash grade . Erlotinib was generally well tolerated . The most frequent erlotinib-related adverse events were rash ( 68 % ) and diarrhea ( 38 % ) . Erlotinib had marginal activity but was generally well tolerated . The safety profile appears more favorable than typically experienced with standard chemotherapeutic agents , which is encouraging in these heavily pretreated patients . Combination of erlotinib with chemotherapy or other targeted agents should be considered . Interferon-alpha promotes the anti-proliferative effect of Erlotinib ( DB00530 ) on human colon cancer cell lines . Interferon-alpha ( IFNalpha ) treatment is associated with up-regulation of epidermal growth factor receptor ( P00533 / P00533 ) expression and marked growth inhibition while maintaining the sensitivity of the target colon cancer cells to epidermal growth factor ( Gut 2004;53:123 ) . We aimed to determine the effect of combining IFNalpha and Erlotinib ( an P00533 / P00533 inhibitor ) on colon cancer cell line growth . Crystal-violet staining and flow cytometry were used to assess cell proliferation and expression of P00533 / P00533 . IFNalpha pre-treatment followed by a combination of IFNalpha plus Erlotinib significantly enhanced the sensitivity of 7/9 of colon cancer cell lines by 7-43 % . This approach may have clinical implications for improving treatment based on targeting of P00533 / P00533 . [ Molecular target-based cancer therapy : epidermal growth factor receptor inhibitors ] . Based on recent progress in cancer biology , numerous molecules that contribute to proliferation , invasion , and metastasis of cancer cells have been identified . The epidermal growth factor receptor ( P00533 ) , a member of cell membrane receptors , is overexpressed by many tumors , and P00533 overexpression correlates with poor prognosis and disease progression . The P00533 is an attractive target for novel anticancer therapy . ZD1839 and DB00530 , highly specific P00533 tyrosine kinase inhibitors , have shown promising antitumor activity against cisplatin-resistant non-small cell lung cancer in phase I and phase II trials . IMC-C225 , a monoclonal antibody against P00533 , has achieved significant disease control in head and neck cancer and colorectal cancer in combination with anticancer agents . These agents are under evaluation in phase III trials . In conclusion , it is expected that P00533 -directed therapies will soon be established as an effective novel treatment for many cancer patients . New targeted therapies in gastrointestinal cancers . Despite surgical , radiotherapeutic , and chemotherapeutic advances , a large proportion of gastrointestinal ( GI ) cancers remain incurable . An improved understanding of the molecular pathogenesis of cancer has promulgated the development of novel agents designed to target critical pathways involved in cancer development and progression . The crucial role of the epidermal growth factor receptor ( P00533 ) in tumor proliferation and the overexpression of P00533 in several GI cancers provides the rationale for targeting and interrupting this key signaling network . P00533 blockade through monoclonal antibodies ( C225 and DB01269 ) and tyrosine kinase inhibitors ( ZD1839 and DB00530 ) has translated into promising evidence of clinical benefit . Ras-mediated signal transduction has been targeted using inhibitors of farnesyl transferase ( R115777 and SCH66336 ) to block the post-translation modification of Ras . Inhibitors of vascular growth factor receptor ( bevacizumab and PTK787 ) and matrix metalloproteinase target the effects of the host environment . P35354 inhibitors in colorectal cancer and STI571 in GI stromal tumors represent novel therapies of interest for these specific GI cancers . Evidence suggests that novel agents can be administered alone or in combination with standard therapies with little additional toxicity . The results of ongoing and future research efforts will clarify the optimal use and survival benefit of targeted therapies for patients with GI malignancies . Role of the JAK- P35610 pathway in protection against myocardial ischemia/reperfusion injury . The Janus kinase ( JAK ) -signal transducers and activators of transcription ( P35610 ) pathway is a stress-responsive mechanism that transduces signals from the cell surface to the nucleus , thereby modulating gene expression . Recent studies have demonstrated that myocardial ischemia and reperfusion induce rapid activation of this pathway . Although the functional consequences of this event remain to be elucidated , there is emerging evidence that JAK- P35610 signaling plays an important role in the development of the cardioprotected phenotype associated with ischemic preconditioning . Specifically , brief episodes of myocardial ischemia/reperfusion activate P23458 and O60674 , followed by recruitment of P42224 and P40763 , resulting in transcriptional upregulation of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) , which then mediate the infarct-sparing effects of the late phase of preconditioning . The present review focuses on this novel cardioprotective role of JAK- P35610 signaling and on its potential exploitation for developing therapeutic strategies aimed at limiting ischemia/reperfusion injury . 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 . Somatic mutations of the epidermal growth factor receptor and non-small-cell lung cancer . Frequent overexpression of epidermal growth factor receptor ( P00533 ) in non-small-cell lung cancer ( NSCLC ) makes P00533 a new therapeutic target . Two specific P00533 tyrosine kinase inhibitors , gefitinib ( ZD1839 , DB00317 ) and erlotinib ( DB00530 , Tarceva ) , have been developed and approved by the US Food and Drug Administration for second-line and third-line treatment of advanced NSCLC . Clinical trials have shown considerable variability in the response rate between different patients with NSCLC , which led to the discovery of somatic P00533 -activating mutations . This brief review summarises the discovery and functional consequences of the mutations , their clinicopathological features and significant implications in the treatment and prognosis of NSCLC . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . A unique structure for epidermal growth factor receptor bound to GW572016 ( DB01259 ) : relationships among protein conformation , inhibitor off-rate , and receptor activity in tumor cells . GW572016 ( DB01259 ) is a tyrosine kinase inhibitor in clinical development for cancer that is a potent dual inhibitor of epidermal growth factor receptor ( P00533 , ErbB-1 ) and ErbB-2 . We determined the crystal structure of P00533 bound to GW572016 . The compound is bound to an inactive-like conformation of P00533 that is very different from the active-like structure bound by the selective P00533 inhibitor DB00530 ( Tarceva ) described previously . Surprisingly , we found that GW572016 has a very slow off-rate from the purified intracellular domains of P00533 and ErbB-2 compared with DB00530 and another P00533 selective inhibitor , ZD-1839 ( DB00317 ) . Treatment of tumor cells with these inhibitors results in down-regulation of receptor tyrosine phosphorylation . We evaluated the duration of the drug effect after washing away free compound and found that the rate of recovery of receptor phosphorylation in the tumor cells reflected the inhibitor off-rate from the purified intracellular domain . The slow off-rate of GW572016 correlates with a prolonged down-regulation of receptor tyrosine phosphorylation in tumor cells . The differences in the off-rates of these drugs and the ability of GW572016 to inhibit ErbB-2 can be explained by the enzyme-inhibitor structures . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . The emerging role of epidermal growth factor receptor inhibitors in ovarian cancer . P00533 ( P00533 ) inhibitors are a new biologically targeted therapy , which may offer new hope in the treatment of patients with advanced or recurrent ovarian cancers . In this review , we summarize and discuss the results of research to date on P00533 inhibitors with particular emphasis on ovarian cancer . We reviewed data identified by searches of MEDLINE , PubMed , and abstracts from the proceedings of the American Society of Clinical Oncology meetings from 1998 to 2006 , with the search terms " Ovarian Cancer, " " P00533 , " " gefitinib , ZD1839 , DB00317 , " " erlotinib , DB00530 , Tarceva, " " DB05424 , " " DB01259 , lapatinib, " " PKI-166, " " Q9Y259 569, " " anti- P00533 antibodies, " " trastuzumab , Herceptin, " " cetuximab , Erbitux , IMC-C225, " " matuzumab , P50402 72000, " " panitumamab , DB01269 , " " pertuzumab , " and " vandetanib , rINN , DB05294 , DB05294 . " Phase II trials of both small molecule inhibitors of P00533 - and antibody-based inhibitors are currently ongoing in ovarian cancer and emerging data suggest that their activity in unselected women with advanced or recurrent ovarian cancer is modest , when utilized as a single agent . It is possible that these agents will be highly effective in smaller subsets of patients whose tumors are dependent on P00533 signaling , perhaps through activating mutations in P00533 or its downstream pathway . Targeted therapy with P00533 inhibitors is an untapped potential resource in the treatment of advanced or recurrent ovarian cancer . Ongoing trials will elucidate the most effective strategies to use these agents individually or in combination with traditional chemotherapeutic agents . Tyrosine phosphorylation and activation of P23458 and P40763 by sublytic C5b-9 complement complex in aortic endothelial cells . The pathway involving Janus kinase ( JAK ) and signal transducers and activators of transcription ( STATs ) plays an important role in differentiation and proliferation of cells initiated by receptor activation . In the present study we identified the JAK and P35610 proteins activated by C5b-9 in human aortic endothelial cells ( AEC ) . P23458 but not O60674 was tyrosine phosphorylated in response to sublytic C5b-9 . P40763 was rapidly tyrosine phosphorylated also by C5b-9 . Pertussis toxin inhibited the C5b-9 induced P23458 activation . However , phosphorylation of P40763 was not inhibited by Pertussis toxin , although C5b-9 induced a time-dependent nuclear translocation of P40763 . These observations indicated that P23458 is phosphorylated by C5b-9 through activation of trimeric G proteins of the Gi/Go family . P04049 and P27361 were also activated by C5b-9 in human AEC in a G protein dependent manner . Therefore , P23458 activity may be involved in activation of P04049 and P27361 via G proteins activated by C5b-9 . This study demonstrates the ability of membrane-inserted C5b-9 to activate P23458 and P40763 proteins , thus defining new signalling pathway by which C5b-9 may regulate gene activation . Inhibition of proliferation , migration , and matrix metalloprotease production in malignant mesothelioma cells by tyrosine kinase inhibitors . The epidermal growth factor receptor ( P00533 ) is expressed in a variety of human solid tumors , including malignant mesothelioma . P00533 has been implicated in regulation of cell proliferation , survival , angiogenesis , and metastasis , making it an ideal target for drug development . ZD1839 ( gefitinib ) and DB00530 ( erlotinib ) are new , low-molecular-weight , P00533 -selective tyrosine kinase ( TK ) inhibitors , whereas DB05424 is a pan- P00533 family TK inhibitor . In the present study , we used ZD1839 , DB00530 , and DB05424 and investigated the effect of these drugs on proliferation , migration , and matrix metalloprotease ( MMP ) production in three malignant mesothelioma cell lines ( M14K , ZL34 , and SPC212 ) . Using [3H]thymidine incorporation , DNA synthesis assay , we found that all three drugs inhibited transforming growth factor-alpha ( TGF-alpha ) -induced cellular proliferation in a dose-dependent manner . In addition , all three drugs induced apoptosis in ZL34 cells as determined by flow cytometry using annexin-V staining . Furthermore , all three drugs inhibited TGF-alpha-induced cell migration ( chemotaxis ) in a dose-dependent manner as determined by Boyden chamber assay . TGF-alpha-induced P14780 production was also inhibited in a dose-dependent manner as determined by gelatin zymography in three cell lines tested . In conclusion , our study demonstrates inhibitory effectiveness of P00533 -TK inhibitors in malignant mesothelioma cells and suggests that these drugs may be an effective treatment strategy for malignant mesothelioma . Targeted therapy in non-small-cell lung cancer . Although treatment of advanced non-small-cell lung cancer has been improved with the availability of such new agents as the taxanes , topoisomerase inhibitors , vinorelbine ( Navelbine ) , and gemcitabine ( Gemzar ) , platinum-based combination therapy has appeared to reach a threshold of therapeutic effectiveness . A paradigm shift in approach to non-small-cell lung cancer and other tumors may be heralded by the development of agents targeting specific biologic pathways in tumor development . Such new agents include antibody epithelial growth factor receptor ( P00533 ) inhibitors ( eg , the monoclonal antibodies trastuzumab [ Herceptin ] and cetuximab [ IMC-C225 , Erbitux ] ) and P00533 tyrosine kinase inhibitors ( eg , ZD1839 [ DB00317 ] and DB00530 ) , angiogenesis inhibitors ( eg , matrix metalloproteinase inhibitors ) , vascular endothelial growth factor ( P15692 ) inhibitors ( eg , monoclonal antibody to P15692 ligand and small-molecule tyrosine kinase ) , and signal transduction inhibitors ( eg , ISIS-3521 , an antisense oligonucleotide to protein kinase C-alpha ) . A number of these agents have entered advanced-phase clinical investigation . It is likely that targeted therapy will have applications in combination with cytotoxic chemotherapy or radiation therapy at all stages of treatment , including maintenance therapy . It is even possible that these new biologic therapies will be used together as rational combinations ( based on pathologic diagnosis ) for advanced non-small-cell lung cancer . Nimotuzumab suppresses epithelial-mesenchymal transition and enhances apoptosis in low-dose UV-C treated salivary adenoid cystic carcinoma cell lines in vitro . Salivary adenoid cystic carcinoma ( SACC ) , which is one of the most common malignant tumors of the salivary glands , is associated with a poor long-term outcome . There are currently few therapeutic options for patients with SACC . Recent studies have shown the potential of the application of ultraviolet-C ( UV-C ) irradiation for the treatment of human cancer . In the present study , we investigated the effects of UV-C in the SACC cell lines SACC-83 and SACC-LM . High-dose UV-C ( 200 J/m ) induced apoptosis and inhibited colony formation significantly . However , low-dose UV-C ( 10 J/m ) , which had little effect on apoptosis and colony formation , increased the ability of migration in SACC cells accompanied by a decrease in P12830 and an increase in vimentin , suggesting the occurrence of epithelial-mesenchymal transition ( EMT ) . Low-dose UV-C ( 10 J/m ) also resulted in upregulation of the phosphorylated forms of epidermal growth factor receptor ( P00533 ) and Akt ( p- P00533 and p-Akt , respectively ) . Pretreatment with Nimotuzumab , an anti- P00533 monoclonal antibody , reversed the EMT as well as upregulation of p- P00533 /p-Akt induced by UV-C . Moreover , Nimotuzumab enhanced UV-C induced apoptosis and inhibition of colony formation . Our results indicate that EMT exerts a protective effect against apoptosis induced by low-dose UV-C . Thus , the combined application of Nimotuzumab and low-dose UV-C in vitro has an advantageous antitumor effect in SACC compared with the application of UV-C alone . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Identification of B cells participated in the mechanism of postmenopausal women osteoporosis using microarray analysis . To further understand the molecular mechanism of lymphocytes B cells in postmenopausal women osteoporosis . Microarray data ( GSE7429 ) were downloaded from Gene Expression Omnibus , in which B cells were separated from the whole blood of postmenopausal women , including 10 with high bone mineral density ( BMD ) and 10 with low BMD . Differentially expressed genes ( DEGs ) between high and low BMD women were identified by Student 's t-test , and P < 0.01 was used as the significant criterion . Functional enrichment analysis was performed for up- and down-regulated DEGs using KEGG , REACTOME , and Gene Ontology ( GO ) databases . Protein-protein interaction network ( PPI ) of up- and down-regulated DEGs was respectively constructed by Cytoscape software using the STRING data . Total of 169 up-regulated and 69 down-regulated DEGs were identified . Functional enrichment analysis indicated that the genes ( Q9BY32 , P31939 , P11172 , P00492 , Q12887 and Q7KZN9 ) might participate in metabolic pathways , Q02779 and P80192 might participate in the activation of P45985 activity , Q12887 and Q7KZN9 might involve in mitochondrial electron transport , and P31939 , P11172 and P00492 might involve in transferase activity . P27361 , Q9BY32 , P31939 , P11172 and P00492 with a higher degree in PPI network were identified . P27361 , Q02779 , P80192 , Q12887 , Q7KZN9 , P31939 , P11172 and P00492 might participate in the pathogenesis of osteoporosis . Invasive Lobular Carcinomas Do Not Express Basal Cytokeratin Markers CK5/6 , CK14 and CK17 . The expression of basal cytokeratin markers CK5/6 in breast carcinomas has been associated with high histological grade and poor clinical outcome . A previous study has shown that CK5/6 can be detected in up to 17 % of invasive lobular carcinomas ( Q9Y4X3 ) . Here we study the expression of three basal cytokeratin markers ( CK5/6 , CK14 , and CK17 ) in 53 Q9Y4X3 cases diagnosed by histology and lack of P12830 expression . Among them , 42 were classic lobular carcinomas , 6 were tubular-lobular carcinoma , and 5 were pleomorphic lobular carcinomas . There was no significant difference among these three groups in patients ' age , tumor size , uni- and multi-focality , expression of ER and PR , lymphovascular invasion , perineural invasion and lymph node metastasis . The only statistically different factor was P04626 over-expression , which was observed only in pleomorphic Q9Y4X3 ( P = 0.0073 ) . None of the 53 cases expressed CK5/6 , CK14 or CK17 ; and 51/53 cases expressed luminal markers CK8 and CK18 , and the two negative cases were both classic lobular carcinoma , with positivity for ER and PR . In conclusion , all 53 cases of Q9Y4X3 failed to show expression by any of the three basal CK markers , suggesting that very few Q9Y4X3 will demonstrate a basal phenotype when assessed by immunohistochemistry ( IHC ) . More studies are needed to investigate molecular classification in lobular carcinoma of the breast . P00533 tyrosine kinase inhibitors in late stage clinical trials . The epidermal growth factor receptor ( P00533 ) is a cell membrane receptor that plays a key role in cancer development and progression . Ligand-activated P00533 -dependent signalling is involved in cell proliferation , apoptosis , angiogenesis and metastatic spread . Targeting the P00533 , therefore , represents a promising molecular approach in cancer treatment . Several anti- P00533 agents are in clinical development . Three drugs are currently in Phase II and III development as single agents , or in combination with other anticancer modalities : IMC-225 ( cetuximab/Erbitux ; ImClone ) , a chimaeric human-mouse monoclonal IgG(1) antibody , which blocks ligand binding and functional activation of the P00533 ; DB00530 ( erlotinib/Tarceva ; Genentech/OSI/Roch ) and ZD1839 ( gefitinib/ DB00317 ; AstraZeneca ) , two small molecule P00533 -selective inhibitors of tyrosine kinase enzymatic activity , which prevent P00533 autophosphorylation and activation . DB00317 is the first P00533 -targeting agent to be registered as an anticancer drug in Japan , in Australia and in the US for the third-line treatment of chemoresistant non-small cell lung cancer ( NSCLC ) patients . This review will focus on the preclinical background and on the results from the first series of clinical trials with these drugs . Furthermore , continuing clinical trials and a series of open clinical issues for the development of optimal strategies of using P00533 -targeting agents will be discussed . Potential role for epidermal growth factor receptor inhibitors in combined-modality therapy for non-small-cell lung cancer . There has been a surge of interest in the translation of discoveries in molecular biology into clinically relevant therapies in the field of hematology/oncology . The epidermal growth factor receptor ( P00533 ) has been a molecular target of significant interest and investigation , and preclinical and clinical studies support a role for targeted therapy in a variety of cancers , including non-small-cell lung cancer ( NSCLC ) via compounds that specifically inhibit P00533 . ZD1839 , IMC-C225 , and DB00530 are the most clinically developed of these compounds . Interestingly , preclinical studies have demonstrated that P00533 inhibitors may have radiation-sensitizing properties , as well as increased cytotoxic activity in combination with chemotherapeutic agents , suggesting a potential role for P00533 inhibitors as an adjunct to the current combined-modality approach for therapy of Stage III NSCLC . Therefore , clinical trials have been proposed and initiated to address the issue of determining the impact of the addition of P00533 inhibitors to the standard combined-modality regimen ( chemotherapy/radiation therapy +/- surgery ) for Stage III NSCLC . This article reviews preclinical and clinical data supporting the role for P00533 inhibitors alone or in combination with chemotherapy/radiation therapy for locally advanced NSCLC . Also , it will provide an overview of ongoing and proposed clinical studies investigating the potential role for P00533 inhibitors in Stage III NSCLC . Targeting epidermal growth factor receptor in lung cancer . Among the most promising agents in clinical development to treat non-small-cell lung cancer ( NSCLC ) are the epidermal growth factor receptor ( P00533 ) targeting agents . A series of recent studies have demonstrated the activity of anti- P00533 targeted therapies for NSCLC . In advanced NSCLC that is refractory to chemotherapy , antitumor responses have been reported with P00533 tyrosine kinase inhibitors ( ZD1839 and DB00530 ) . The role of ZD1839 and DB00530 as possible additions to standard chemotherapy in the first-line setting has also been evaluated , and the studies conducted to date should respond to the question of whether these compounds could provide a survival benefit . Other areas of research involve looking at the role of P00533 tyrosine kinase inhibitors in the neoadjuvant treatment of stage III NSCLC and the planning of chemoprevention studies . These exciting results and plans are further complemented by an emerging number of compounds in clinical development , including both monoclonal antibodies ( ie , IMC-C225 ) and other tyrosine kinase inhibitors , directed at the P00533 . Targeting epidermal growth factor receptor : novel therapeutics in the management of cancer . Overexpression of epidermal growth factor receptor ( P00533 ) in epithelial tumors , including head and neck , lung , breast , colon and other solid tumors , has frequently been correlated with poor prognosis , thus stimulating efforts to develop new cancer therapies that target P00533 . Monoclonal antibodies and tyrosine kinase inhibitors specifically targeting P00533 are the most well-studied and hold substantial promise of success . Several compounds of monoclonal antibodies and tyrosine kinase inhibitors targeting P00533 have been studied and clinical trials are now underway to test the safety and efficacy of these targeting strategies in several human tumors . This review will address each of these agents alone or in combination with radiation or chemotherapy and highlight some of these promising developments . Cetuximab ( Erbitux ) is being evaluated in combination with radiation or chemotherapy in Phase III trials . Other compounds such as h-R3 , DB01269 , P50402 -55900 and ICR-62 have proved to be effective in targeting malignant cells alone or in combination with traditional therapies . Tyrosine kinase inhibitors targeting the intracellular domain of P00533 , including ZD-1839 ( gefitinib , DB00317 ) , DB00530 ( Erlotinib/Tarceva ) , PD-153053 , PD-168393 and DB05424 , have been studied in clinical setting alone or in combination with radiation or chemotherapy . ZD-1839 is being studied in a Phase III trial in patients with advanced non-small cell lung cancer . P00533 targeted treatment by monoclonal antibodies and tyrosine kinase inhibitors have been proven to sensitize tumor cells to the effects of chemotherapy and radiation therapy . The synergistic activities and nonoverlapping toxicities of these compounds allow concomitant administration with cytotoxic therapy . Challenges of evaluating P00533 targeted agents exist in selecting the optimal dosages and determining long-term toxicity . 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 . Development of the epidermal growth factor receptor inhibitor Tarceva ( DB00530 ) . The epidermal growth factor receptor ( P00533 ) is a transmembrane receptor involved in the regulation of a complex array of essential biological processes such as cell proliferation and survival . Dysregulation of P00533 signaling network has been frequently reported in multiple human cancers and has been associated with the processes of tumor development , growth , proliferation , metastasis and angiogenesis . Inhibition of the P00533 was associated with antitumor effects in preclinical models . On the bases of these data , therapeutics targeting the P00533 were explore in clinical trials . Tarceva ( DB00530 , OSI Pharmaceuticals , Uniondale , NY ) is a small molecule selective inhibitor of the P00533 tyrosine kinase ( TK ) . In preclinical studies , Tarceva inhibited the phosphorylation of the P00533 in a dose and concentration dependent manner resulting in cell cycle arrest and induction of apoptosis . In in vivo studies , the agent caused tumor growth inhibition and shoved synergistic effects when combined with conventional chemotherapy . Subsequent single agent phase I studies and phase I studies in combination with chemotherapy demonstrated that the agent has a good safety profile and induced tumor growth inhibition in a substantial number of patients with a variety of different solid tumor . Preliminary report from phase II studies confirmed the excellent tolerability of Tarceva as well as showed encouraging preliminary activity . Phase III studies have either been completed or are ongoing in several tumor types such as lung cancer and pancreatic cancer . In summary , Tarceva is a novel inhibitor of the P00533 TK which has shown promising activity in initial studies and is currently undergoing full development as an anticancer drug . Structure of the epidermal growth factor receptor kinase domain alone and in complex with a 4-anilinoquinazoline inhibitor . The crystal structure of the kinase domain from the epidermal growth factor receptor ( EGFRK ) including forty amino acids from the carboxyl-terminal tail has been determined to 2.6-A resolution , both with and without an EGFRK-specific inhibitor currently in Phase III clinical trials as an anti-cancer agent , erlotinib ( DB00530 , CP-358,774 , Tarceva(TM) ) . The P00533 family members are distinguished from all other known receptor tyrosine kinases in possessing constitutive kinase activity without a phosphorylation event within their kinase domains . Despite its lack of phosphorylation , we find that the EGFRK activation loop adopts a conformation similar to that of the phosphorylated active form of the kinase domain from the insulin receptor . Surprisingly , key residues of a putative dimerization motif lying between the EGFRK domain and carboxyl-terminal substrate docking sites are found in close contact with the kinase domain . Significant intermolecular contacts involving the carboxyl-terminal tail are discussed with respect to receptor oligomerization . A phase II study of erlotinib ( DB00530 ) given in combination with carboplatin in patients with recurrent epithelial ovarian cancer ( NCIC CTG IND.149 ) . OBJECTIVES : Approximately 50 % of ovarian cancers have elevated levels of epidermal growth factor receptor ( P00533 ) which correlates with a poor prognosis . Preclinical evidence suggests that P00533 tyrosine kinase inhibitors ( TKIs ) , such as erlotinib ( DB00530 ) , may potentiate the anti-tumour effects of cytotoxic agents , including carboplatin . Blocking P00533 could thus potentially reverse drug resistance . The primary objective of the study was to assess the response rate to the addition of erlotinib in patients with recurrent ovarian cancer who were receiving carboplatin . METHODS : Patients enrolled on this study had either local or advanced recurrent ovarian cancer with measurable disease . They may have had up to 2 prior chemotherapy regimens , one of which must have contained platinum , and they must have responded to prior platinum therapy . Patients were stratified by platinum sensitivity and were treated with erlotinib 150 mg daily on a continuous dosing schedule , and carboplatin at an AUC of 5 every 21 days . RESULTS : Fifty patients with recurrent ovarian cancer entered the study , 33 in the platinum-sensitive arm and 17 in the platinum-resistant arm . Of patients evaluable for response , there were 14 partial responses ( PR ) of 30 evaluable for response ( 57 % objective response rate ( ORR ) ) in the platinum-sensitive arm , and 1 PR of 14 evaluable for response ( 7 % ORR ) in the platinum-resistant arm . CONCLUSIONS : The combination of erlotinib and carboplatin was active in patients with platinum-sensitive disease , but not in platinum-resistant disease . The toxicities seen were those expected with carboplatin and erlotinib . Development of the epidermal growth factor receptor inhibitor DB00530 . The epidermal growth factor receptor ( P00533 ) is a transmembrane receptor involved in the regulation of a complex array of essential biological processes such as cell proliferation and survival . Dysregulation of the P00533 signaling network has been frequently reported in multiple human cancers and has been associated with the processes of tumor development , growth , proliferation , metastasis , and angiogenesis . Inhibition of the P00533 was associated with antitumor effects in preclinical models . On the basis of these data , therapeutics targeting the P00533 were explored in clinical trials . DB00530 is a small-molecule selective inhibitor of the P00533 tyrosine kinase . In preclinical studies , DB00530 inhibited the phosphorylation of the P00533 in a dose-dependent and concentration-dependent manner resulting in cell cycle arrest and induction of apoptosis . In in vivo studies , this agent caused tumor growth inhibition and showed synergistic effects when combined with conventional chemotherapy . Subsequent single-agent phase I studies and phase I studies in combination with chemotherapy showed that the agent has a good safety profile and induced tumor growth inhibition in a substantial number of patients with a variety of different solid tumors . Preliminary reports from phase II studies confirmed the excellent tolerability of DB00530 and showed encouraging preliminary activity . Phase III studies have either been completed or are ongoing in several tumor types such as lung cancer and pancreatic cancer . In summary , DB00530 is a novel inhibitor of the P00533 tyrosine kinase that has shown promising activity in initial studies and is currently undergoing full development as an anticancer drug . Functional role of wogonin in anti-angiogenesis . Constitutive activation of the Janus kinase ( JAK ) /signal transducer and activator of transcription ( P35610 ) pathway occurs commonly in cancer cells and endothelial cells , and contributes to angiogenesis . Wogonin is a compound with many biologically relevant properties . We previously reported that wogonin blocked P05231 -induced angiogenesis through suppression of P15692 expression , an important regulator of angiogenesis . However , the pathway involved in the suppressive effect of wogonin on P05231 -induced P15692 has not been completely clarified . This study aimed to investigate the molecular mechanisms participating in the suppression of wogonin on P05231 -induced P15692 in vitro , focusing on IL-6R/ P23458 / P40763 / P15692 pathway . Both P40763 siRNA and wogonin treatment resulted in an abolition of the expression of P15692 . Moreover , our data revealed that wogonin treatment after P40763 knock-down did not further suppress P15692 expression . The addition of IL-6R siRNA or wogonin resulted in a decrease in the expression level of the phosphorylated P23458 protein . Furthermore , wogonin significantly decreased the amount of phosphorylated P40763 . Finally , by EMSA , wogonin suppressed P05231 -induced P40763 binding activity in a concentration-dependent manner . Taken together , our results show that wogonin suppresses P05231 -induced P15692 by modulating the IL-6R/ P23458 / P40763 signaling pathway . Based on this study , we suggest that wogonin may provide a new potential therapeutic option for treatment of P05231 -related pathological angiogenesis . Biologically targeted treatment of non-small-cell lung cancer : focus on epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) has emerged in recent years as a key target of molecular therapy for solid tumors . The postembryonic role of P00533 is normally limited . In cancer , however , abnormal P00533 -tyrosine kinase ( TK ) activity plays a central role in many of the processes involved in tumor progression , such as proliferation , angiogenesis , invasiveness , decreased apoptosis , and loss of differentiation . Several different approaches have been taken to inhibit P00533 -mediated activity in tumor cells , including monoclonal antibodies directed at the ligand-binding portion of the P00533 and small-molecule agents that directly inhibit the intracellular TK domain of P00533 . Two of these TK inhibitors , gefitinib and erlotinib ( DB00530 , Tarceva ) , have shown antitumor activity and good tolerability across several tumor types in early dose-finding clinical trials , particularly for non-small-cell lung cancer ( NSCLC ) . In heavily pretreated patients with advanced NSCLC , gefitinib showed clinically significant tumor responses and symptom relief with good tolerability . Based on these results , gefitinib has now been approved for the third-line treatment of advanced NSCLC . The use of gefitinib in standard treatment programs or combined with other molecular targeted agents may substantially improve the outlook for patients with NSCLC or other types of solid tumors Why the epidermal growth factor receptor ? The rationale for cancer therapy . There is a need for new , selective anticancer agents that differentiate between malignant and nonmalignant cells . The benefits of such agents would include a higher therapeutic index and lower toxicity than conventional therapies . Although expressed in nonmalignant cells , the epidermal growth factor receptor ( P00533 ) is highly expressed in a variety of tumors , and its expression correlates with poor response to treatment , disease progression , and poor survival . Evidence for a role for the P00533 in the inhibition and pathogenesis of various cancers has led to the rational design and development of agents that selectively target this receptor . Activation of the P00533 signaling pathway in cancer cells has been linked with increased cell proliferation , angiogenesis , and metastasis , and decreased apoptosis . Preclinical data show that anti- P00533 therapies can inhibit these effects in vitro and in vivo . In addition , preclinical data confirm that many such agents have the potential to increase the effectiveness of current cytotoxic agents . Following accelerated drug development programs , phase III trials are now under way for a number of P00533 -targeted therapies , including the monoclonal antibody IMC-C225 and the P00533 -tyrosine kinase inhibitors ZD1839 ( DB00317 ) and DB00530 . Thus , the rationale for P00533 -targeted approaches to cancer treatment is apparent and now well established , and there is increasing evidence that they may represent a significant contribution to cancer therapy . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . P25101 antagonists prevent the development of pulmonary emphysema in rats . We hypothesised that endothelin ( ET ) -1 plays an important role in the pathogenesis of emphysema . We attempted to apply ET-1 receptor antagonists to demonstrate and further elucidate the molecular pathogenesis pathways through which ET-1 may cause emphysematous changes . Sprague-Dawley rats were divided into four groups : control , cigarette smoke extract ( CSE ) , CSE+BQ-123 ( a selective endothelin receptor type A ( ET(A) ) antagonist ) and CSE+ DB00559 ( a mixed ET(A)/ET(B) receptor antagonist ) . The CSE was injected intraperitoneally once a week for 3 weeks , and BQ-123 or DB00559 was administered daily for the same duration . The expression of ET(A) receptor , apoptosis index , caspase-3 activity , matrix metalloproteinase ( MMP ) -2 and P14780 activity , and tumour necrosis factor ( P01375 ) -alpha and interleukin ( IL ) -1beta concentrations were measured in the lung tissue . The ET-1 levels and antioxidant activity were measured in the serum . Both BQ-123 and DB00559 prevented the development of CSE-induced emphysema , blocked the expression of ET(A) receptor , inhibited pulmonary apoptosis , inactivated P08253 and P14780 activities in the lung tissues , reduced the concentrations of inflammatory cytokines P01375 and IL-1beta , and improved the biological antioxidant activity in the serum . Emphysema development is suppressed by ET-1 receptor antagonists . ET-1 may cause emphysematous changes through molecular pathogenesis pathways involving apoptosis , proteinase and antiproteinase imbalance , inflammation and oxidative stress . Ventilation-induced increases in P00533 ligand mRNA are not altered by intra-amniotic LPS or ureaplasma in preterm lambs . Chorioamnionitis and mechanical ventilation are associated with bronchopulmonary dysplasia ( BPD ) in preterm infants . Mechanical ventilation at birth activates both inflammatory and acute phase responses . These responses can be partially modulated by previous exposure to intra-amniotic ( IA ) LPS or Ureaplasma parvum ( UP ) . P00533 ( P00533 ) ligands participate in lung development , and angiotensin converting enzyme ( P12821 ) 1 and Q9BYF1 contribute to lung inflammation . We asked whether brief mechanical ventilation at birth altered P00533 and P12821 pathways and if antenatal exposure to IA LPS or UP could modulate these effects . Ewes were exposed to IA injections of UP , LPS or saline multiple days prior to preterm delivery at 85 % gestation . Lambs were either immediately euthanized or mechanically ventilated for 2 to 3 hr . IA UP and LPS cause modest changes in the P00533 ligands amphiregulin ( P15514 ) , epiregulin ( O14944 ) , heparin binding epidermal growth factor ( HB- P01133 ) , and betacellulin ( P35070 ) mRNA expression . Mechanical ventilation greatly increased mRNA expression of P15514 , O14944 , and HB- P01133 , with no additional increases resulting from IA LPS or UP . With ventilation P15514 and O14944 mRNA localized to cells in terminal airspace . P00533 mRNA also increased with mechanical ventilation . IA UP and LPS decreased ACE1 mRNA and increased Q9BYF1 mRNA , resulting in a 4 fold change in the ACE1/ Q9BYF1 ratio . Mechanical ventilation with large tidal volumes increased both ACE1 and Q9BYF1 expression . The alterations seen in P12821 with IA exposures and P00533 pathways with mechanical ventilation may contribute to the development of BPD in preterm infants . Erlotinib ( Tarceva , DB00530 ) antagonizes DB00171 -binding cassette subfamily B member 1 and DB00171 -binding cassette subfamily G member 2-mediated drug resistance . It has been reported that gefitinib , an epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor ( TKI ) , has the ability to modulate the function of certain DB00171 -binding cassette ( DB01048 ) transporters and to reverse ABC subfamily B member 1 ( P08183 ; P-glycoprotein ) - and ABC subfamily G member 2 ( Q9UNQ0 ; breast cancer resistance protein/mitoxantrone resistance protein ) -mediated multidrug resistance ( MDR ) in cancer cells . However , it is unknown whether other P00533 TKIs have effects similar to that of gefitinib . In the present study , we have investigated the interaction of another P00533 TKI , erlotinib , with selected ABC drug transporters . Our findings show that erlotinib significantly potentiated the sensitivity of established P08183 or Q9UNQ0 substrates and increased the accumulation of paclitaxel or mitoxantrone in P08183 - or Q9UNQ0 -overexpressing cells . Furthermore , erlotinib did not significantly alter the sensitivity of non- P08183 or non- Q9UNQ0 substrates in all cells and was unable to reverse MRP1-mediated MDR and had no effect on the parental cells . However , erlotinib remarkably inhibited the transport of E(2)17 beta G and methotrexate by Q9UNQ0 . In addition , the results of ATPase assays show that erlotinib stimulated the ATPase activity of both P08183 and Q9UNQ0 . Interestingly , erlotinib slightly inhibited the photolabeling of P08183 with [(125)I]iodoarylazidoprazosin ( IAAP ) at high concentration , but it did not inhibit the photolabeling of Q9UNQ0 with IAAP . Overall , we conclude that erlotinib reverses P08183 - and Q9UNQ0 -mediated MDR in cancer cells through direct inhibition of the drug efflux function of P08183 and Q9UNQ0 . These findings may be useful for cancer combinational therapy with erlotinib in the clinic . 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 An P01133 receptor inhibitor induces P10826 expression in breast and ovarian cancer cells . Inhibition of the epidermal growth factor ( P01133 ) -receptor ( P00533 ) has become a promising anticancer treatment strategy . In addition , application of retinoids yields encouraging results for cancer prevention and therapy . Many tumors express no or low amounts of retinoic acid receptor-beta2 ( RAR-beta2 ) due to epigenetic silencing via DNA hypermethylation . RAR-beta2 is the main mediator of the antiproliferative effect of retinoids . RAR-beta2 re-expression causes reversal of transformation , cell cycle arrest , and restoration of retinoid sensitivity . RAR-beta2 is thus a tumor suppressor . Western blotting , colorimetric in vitro cell proliferation assays , and reverse transcription-polymerase chain reaction demonstrated that the P00533 inhibitor PD153035 not only blocked activation of P00533 and inhibited cell growth , but also stimulated P10826 expression in MDA-MB-468 breast and OVCAR-3 ovarian carcinoma cells . Upregulation of P10826 by PD153035 was confirmed by real-time reverse transcription-polymerase chain reaction . In contrast , expression of other retinoid receptors and of estrogen receptor-alpha was not affected . PD153035-mediated re-induction of P10826 was associated with demethylation of the RAR-beta2 gene promoter P2 as demonstrated by methylation-specific polymerase chain reaction . These novel results on the ErbB/retinoid receptor cross-talk may be useful for designing future anticancer combination regimens . P00533 inhibitors , gefitinib and erlotinib ( Tarceva , DB00530 ) , in the treatment of bronchioloalveolar carcinoma . Tamoxifen enhances erlotinib-induced cytotoxicity through down-regulating AKT-mediated thymidine phosphorylase expression in human non-small-cell lung cancer cells . Tamoxifen is a triphenylethylene nonsteroidal estrogen receptor ( ER ) antagonist used worldwide as an adjuvant hormone therapeutic agent in the treatment of breast cancer . However , the molecular mechanism of tamoxifen-induced cytotoxicity in non-small cell lung cancer ( NSCLC ) cells has not been identified . P19971 ( TP ) is an enzyme of the pyrimidine salvage pathway which is upregulated in cancers . In this study , tamoxifen treatment inhibited cell survival in two NSCLC cells , H520 and H1975 . Treatment with tamoxifen decreased TP mRNA and protein levels through AKT inactivation . Furthermore , expression of constitutively active AKT ( AKT-CA ) vectors significantly rescued the decreased TP protein and mRNA levels in tamoxifen-treated NSCLC cells . In contrast , combination treatment with PI3K inhibitors ( LY294002 or wortmannin ) and tamoxifen further decreased the TP expression and cell viability of NSCLC cells . Knocking down TP expression by transfection with small interfering RNA of TP enhanced the cytotoxicity and cell growth inhibition of tamoxifen . Erlotinib ( Tarceva , DB00530 ) , an orally available small molecular inhibitor of epidermal growth factor receptor ( P00533 ) tyrosine kinase , is approved for clinical treatment of NSCLC . Compared to a single agent alone , tamoxifen combined with erlotinib resulted in cytotoxicity and cell growth inhibition synergistically in NSCLC cells , accompanied with reduced activation of phospho-AKT and phospho- P27361 /2 , and reduced TP protein levels . These findings may have implications for the rational design of future drug regimens incorporating tamoxifen and erlotinib for the treatment of NSCLC . Phase II clinical trial data with the epidermal growth factor receptor tyrosine kinase inhibitor erlotinib ( DB00530 ) in non-small-cell lung cancer . Erlotinib ( DB00530 ; Tarceva ) is an orally available , highly specific epidermal growth factor receptor ( P00533 ) tyrosine kinase inhibitor . The results of 3 phase II studies with erlotinib in non-small-cell lung cancer ( NSCLC ) are reviewed herein : ( 1 ) in patients with chemotherapy-resistant , P00533 / P00533 -expressing NSCLC of all histologies , ( 2 ) in patients with bronchoalveolar carcinoma previously untreated or treated with chemotherapy , and ( 3 ) as first-line therapy in elderly patients with NSCLC of all histologies . These studies have evaluated tumor response , survival , and symptom improvement . Erlotinib was given as an oral , continuous daily dose of 150 mg . The drug was well tolerated ; drug-related cutaneous rash and diarrhea were observed in approximately two thirds of patients . Withdrawals caused by toxicity were rare . The response rates were 12.3 % , 25 % , and 13.3 % , respectively . Mature survival data are available for the first trial . The median survival was 8.4 months , and the 1-year survival rate was 40 % . All responding patients in the first and second trials presented skin rash . In addition , survival correlated with the occurrence and severity of rash in the first trial . No data on the correlation between rash and survival are available for the second and third trials . Erlotinib is active and well tolerated in patients with NSCLC as first- and second-line therapy . Cutaneous rash appears to be a surrogate marker of clinical benefit , but this finding needs to be confirmed in ongoing and future studies . DB00530 OSI Pharmaceuticals . DB00530 ( formerly CP-358774 ) , a quinazoline derivative , is an orally active epidermal growth factor receptor ( P00533 ) inhibitor which was originally under joint development by Pfizer and OSI Pharmaceuticals ( formerly Oncogene Science ) for the potential treatment of cancer ( eg , ovarian , non-small cell lung cancer ( NSCLC ) and head and neck ) . It is being evaluated in phase II trials [ 304305 ] , [ 372201 ] . On 8 January 2001 , OSI announced that it had signed an agreement with Roche and Genentech for the global co-development and marketing of DB00530 . The agreement with Genentech covers the United States , that with Roche the rest of the world [ 395371 ] , [ 395526 ] . In June 2000 , OSI gained all development and marketing rights for DB00530 following Pfizer 's merger with Warner-Lambert [ 371439 ] . In September 2000 , Pfizer transferred the IND dossierfor DB00530 to OSI ahead of the timeline agreed in the June 2000 development and marketing rights agreement [ 383786 ] . The phase II trials will assess DB00530 both as a single agent and in combination with existing chemotherapy regimens [ 347783 ] . Phase III trials are expected to be initiated in 2001 [ 347783 ] . In October 2000 , Lehman Brothers predicted that DB00530 would move into pivotal trials in thefirst half of 2001 and that the drug would be launched in 2003 . The analysts also estimated worldwide sales of US $ 66 million , $ 285 million and $ 461 million in 2003 , 2004 and 2005 , respectively , and peak sales in excess of US $ 500 million [ 395189 ] . Rationale and clinical validation of epidermal growth factor receptor as a target in the treatment of head and neck cancer . Recurrent/metastatic head and neck cancer is an area of high , unmet treatment need . There is a strong rationale for targeting the epidermal growth factor receptor ( P00533 ) in head and neck cancer as most of these tumors express high levels of P00533 relative to normal tissue , with high expression correlating with poor patient outcome . This rationale has been validated in extensive preclinical studies . Two small molecules with P00533 inhibitory activity , gefitinib ( ' DB00317 ' , ZD1839 ) and erlotinib ( ' Tarceva ' , DB00530 ) , and a humanized monoclonal antibody against the P00533 extracellular domain , cetuximab ( ' Erbitux ' , C225 ) , are in clinical trials for advanced head and neck cancer . The initial results of these trials are promising . Gefitinib and erlotinib show activity as monotherapy in patients with recurrent or metastatic head and neck cancer , and have an acceptable safety profile compared with conventional chemotherapy . Gefitinib , which can be given at doses below the maximum tolerated dose , is associated with slightly lower rates of adverse events than erlotinib , which is dosed at the maximum tolerated dose . Combinations of cetuximab with radiotherapy or platinum-based chemotherapy have also shown activity in phase I/II studies . Both gefitinib and cetuximab have entered phase III studies . The results of these trials , which will mature over the next few years , will help determine the optimal use of P00533 agents in head and neck cancers . In vitro effects and ex vivo binding of an P00533 -specific immunotoxin on rhabdomyosarcoma cells . PURPOSE : Rhabdomyosarcoma ( RMS ) is a rare and aggressive soft tissue sarcoma with limited treatment options and a high failure rate during standard therapy . New therapeutic strategies based on targeted immunotherapy are therefore much in demand . The epidermal growth factor receptor ( P00533 ) has all the characteristics of an ideal target . It is overexpressed in up to 80 % of embryonal RMS and up to 50 % of alveolar RMS tumors . We therefore tested the activity of the P00533 -specific recombinant immunotoxin ( IT ) 425(scFv)- P25101 ' against P00533 (+) RMS cells in vitro and ex vivo . METHODS : We tested the specific binding and internalization behavior of 425(scFv)- P25101 ' in RMS cell lines in vitro by flow cytometry , compared to the corresponding imaging probe 425(scFv)- P60880 monitored by live cell imaging . The cytotoxic activity of 425(scFv)- P25101 ' was tested using cell viability and apoptosis assays . Specific binding of the IT was confirmed on formalin-fixed paraffin-embedded tissue samples from two RMS patients . RESULTS : We confirmed the specific binding of 425(scFv)- P25101 ' to RMS cells in vitro and ex vivo . Both the IT and the corresponding imaging probe were rapidly internalized . The IT killed P00533 (+) RMS cells in a dose-dependent manner , while showing no effect against control cells . It showed specific apoptotic activity against one selected RMS cell line . CONCLUSIONS : This is the first study showing the promising therapeutic potential of a recombinant , P00533 -targeting , P25101 '-based IT on RMS cells . We confirmed the selective killing with IC50 values of up to 50 pM , and immunohistochemical staining confirmed the specific ex vivo binding to primary RMS material . Inactivation of Akt by the epidermal growth factor receptor inhibitor erlotinib is mediated by HER-3 in pancreatic and colorectal tumor cell lines and contributes to erlotinib sensitivity . Signaling through the receptor for epidermal growth factor receptor ( P00533 ) is frequently deregulated in solid tumors . Erlotinib ( Tarceva , DB00530 , OSI Pharmaceuticals , Inc. , Melville , NY ) is a low molecular weight , orally bioavailable inhibitor of the P00533 that has been approved for both non-small cell lung cancer and pancreatic cancers . Previous studies have indicated that sensitivity to P00533 antagonists correlated with HER-3 signaling for non-small cell lung cancer . Herein , we have sought to understand the signaling pathways that mediate erlotinib sensitivity for pancreatic and colorectal cancers . In a panel of 12 pancreatic tumor cell lines , we find that P00533 is coexpressed with HER-3 in all cell lines sensitive to erlotinib but not in insensitive cell lines . Erlotinib can block HER-3 phosphorylation in these sensitive cell lines , suggesting that HER-3 is transactivated by P00533 . Knockdown of HER-3 in BxPC3 , an erlotinib-sensitive pancreatic tumor cell line , results in inhibition of the phosphorylation for both Akt and S6 and is associated with a decrease in cell proliferation and reduced sensitivity to erlotinib . Therefore , P00533 transactivation of HER-3 mediates Akt signaling and can contribute to erlotinib sensitivity for pancreatic tumors . We extended our analysis to a panel of 13 colorectal tumor cell lines and find that , like pancreatic , HER-3 is coexpressed with P00533 in the most erlotinib-sensitive cell lines but not in erlotinib-insensitive cell lines . These studies suggest that HER-3 could be used as a biomarker to select patients who are most likely to respond to erlotinib therapy . Combination treatment of glioblastoma multiforme cell lines with the anti-malarial artesunate and the epidermal growth factor receptor tyrosine kinase inhibitor DB00530 . New drugs and combination modalities for otherwise non-responsive brain tumors are urgently required . The anti-malarial artesunate ( O00253 ) and the P00533 tyrosine kinase inhibitor DB00530 reveal profound cytotoxic activity . The effectiveness of a combination treatment and the underlying molecular determinants of cellular response are unknown . In the present investigation , we studied O00253 and DB00530 in glioblastoma multiforme ( GBM ) cell lines . Supra-additive inhibition of cell growth was observed in U-87MG.DeltaEGFR cells transduced with a deletion-mutant constitutively active P00533 gene , while additive effects were present in cells transduced with wild-type P00533 ( U-87MG.WT-2N ) , kinase-deficient P00533 ( U-87MG.DK-2N ) , mock vector controls ( U-87MG.LUX ) , or non-transduced parental U-87MG cells . Among nine other non-transduced GBM cell lines , supra-additive effects were found in two cell lines ( G-210GM , G-599GM ) , while O00253 and DB00530 acted in an additive manner in the other seven cell lines ( G-211GM , G-750GM , G-1163GM , G-1187GM , G-1265GM , G-1301GM , and G-1408GM ) . Sub-additive or antagonistic effects were not observed . Genomic gains and losses of genetic material in the non-transduced cell lines as assessed by comparative genomic hybridization were correlated with the IC(50) values for O00253 and DB00530 and subsequently subjected to hierarchical cluster analysis and cluster image mapping . A genomic profile of imbalances was detected that predicted cellular response to O00253 and DB00530 . The genes located at the genomic imbalances of interest may serve as candidate resistance genes of GBM cells towards O00253 and DB00530 . In conclusion , the combination treatment of O00253 and DB00530 resulted in an increased growth inhibition of GBM cell lines as compared to each drug alone . Gefitinib ( ' DB00317 ' , ZD1839 ) and new epidermal growth factor receptor inhibitors . The epidermal growth factor receptor ( P00533 ) is a promising target for cancer therapy and a number of P00533 -targeted agents have been developed . Those most advanced in development are the P00533 tyrosine kinase inhibitors gefitinib ( ' DB00317 ' , ZD1839 ) and erlotinib ( ' Tarceva ' , DB00530 ) , and the monoclonal antibody cetuximab ( ' Erbitux ' , IMC-C225 ) . This review provides a clinical overview of these agents , highlighting their antitumour activities in different tumour types . P00533 -targeted agents are generally well tolerated and are not typically associated with the severe adverse events often seen with cytotoxic chemotherapy . Gefitinib is the agent with the most extensive clinical experience , particularly in non-small-cell lung cancer ( NSCLC ) . Recently , gefitinib became the first-approved P00533 -targeted agent , for use in patients with previously treated advanced NSCLC in Japan , the USA and other countries . Further studies are required to explore the full potential of these novel agents either as monotherapy or combination therapy . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . A transgenic platform for testing drugs intended for reversal of cardiac remodeling identifies a novel 11βHSD1 inhibitor rescuing hypertrophy independently of re-vascularization . RATIONALE : Rescuing adverse myocardial remodeling is an unmet clinical goal and , correspondingly , pharmacological means for its intended reversal are urgently needed . OBJECTIVES : To harness a newly-developed experimental model recapitulating progressive heart failure development for the discovery of new drugs capable of reversing adverse remodeling . METHODS AND RESULTS : A P15692 -based conditional transgenic system was employed in which an induced perfusion deficit and a resultant compromised cardiac function lead to progressive remodeling and eventually heart failure . Ability of candidate drugs administered at sequential remodeling stages to reverse hypertrophy , enlarged LV size and improve cardiac function was monitored . Arguing for clinical relevance of the experimental system , clinically-used drugs operating on the P00797 -Angiotensin- DB04630 -System ( RAAS ) , namely , the P12821 inhibitor Enalapril and the direct renin inhibitor Aliskerin fully reversed remodeling . Remodeling reversal by these drugs was not accompanied by neovascularization and reached a point-of-no-return . Similarly , the PPARγ agonist Pioglitazone was proven capable of reversing all aspects of cardiac remodeling without affecting the vasculature . Extending the arsenal of remodeling-reversing drugs to pathways other than RAAS , a specific inhibitor of 11β-hydroxy-steroid dehydrogenase type 1 ( 11β HSD1 ) , a key enzyme required for generating active glucocorticoids , fully rescued myocardial hypertrophy . This was associated with mitigating the hypertrophy-associated gene signature , including reversing the myosin heavy chain isoform switch but in a pattern distinguishable from that associated with neovascularization-induced reversal . CONCLUSIONS : A system was developed suitable for identifying novel remodeling-reversing drugs operating in different pathways and for gaining insights into their mechanisms of action , exemplified here by uncoupling their vascular affects . Mutant epidermal growth factor receptor up-regulates molecular effectors of tumor invasion . The gene most commonly altered in human glioblastomas is the epidermalgrowth factor receptor ( P00533 ) . We profiled transcripts induced by mutantEGFR to better understand its role in tumor progression . The pattern found suggested enhanced tumor invasion . The highly induced genes included extracellular matrix components , metalloproteases , and a serine protease . We confirmed that mutant P00533 did make glioblastoma cells both more motile and invasive using in vitro assays . Furthermore , inhibitors of P00533 ( DB00530 and Tyrphostin AG1478 ) selectively down-regulated these molecular effectors in glioblastoma cells , eliminating enhanced invasion . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . DB00227 overcomes gefitinib resistance in human non-small cell lung cancer cells with K-Ras mutations . DB00227 is a 3-hydroxy-3-methylglutaryl-coenzyme A ( HMG- DB01992 ) reductase inhibitor . Its inhibitory action on P04035 leads to depletion of isoprenoids , which inhibits post-translational modification of DB01367 . In this study , we investigated the effect of combining lovastatin with gefitinib on gefitinib-resistant human non-small cell lung cancer ( NSCLC ) cell lines with K-Ras mutations . Antitumor effects were measured by growth inhibition and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay . Effects on apoptosis were determined by flow cytometry , DNA fragmentation , and immunoblots . Protein levels of DB01367 , AKT/pAKT , and RAF/ P27361 /2 in cancer cells were analyzed by immunoblot . Compared with gefitinib alone , a combination of gefitinib with lovastatin showed significantly enhanced cell growth inhibition and cytotoxicity in gefitinib-resistant A549 and NCI-H460 human NSCLC cells . In addition , lovastatin combination treatment significantly increased gefitinib-related apoptosis , as determined by fluorescence microscopy and flow cytometric analysis . These effects correlated with up-regulation of cleaved caspase-3 , poly ( ADP-ribose ) polymerase ( PARP ) , and Bax and down-regulation of Bcl-2 . The combination of lovastatin and gefitinib effectively down-regulated DB01367 protein and suppressed the phosphorylation of RAF , P27361 /2 , AKT , and P00533 in both cell lines . Taken together , these results suggest lovastatin can overcome gefitinib resistance , in NSCLC cells with K-Ras mutations , by down regulation of DB01367 protein , which leads to inhibition of both RAF/ P29323 and AKT pathways . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . P10275 controls P00533 and P04626 gene expression at different levels in prostate cancer cell lines . P00533 or P04626 contributes to prostate cancer ( PCa ) progression by activating the androgen receptor ( AR ) in hormone-poor conditions . Here , we investigated the mechanisms by which androgens regulate P00533 and P04626 expression in PCa cells . In steroid-depleted medium ( SDM ) , P00533 protein was less abundant in androgen-sensitive LNCaP than in androgen ablation-resistant 22Rv1 cells , whereas transcript levels were similar . DB02901 ( DB02901 ) treatment increased both P00533 mRNA and protein levels and stimulated RNA polymerase II recruitment to the P00533 gene promoter , whereas it decreased P04626 transcript and protein levels in LNCaP cells . DB02901 altered neither P00533 or P04626 levels nor the abundance of prostate-specific antigen ( PSA ) , TMEPA1 , or O15393 mRNAs in 22Rv1 cells , which express the full-length and a shorter AR isoform deleted from the COOH-terminal domain ( ARDeltaCTD ) . The contribution of both AR isoforms to the expression of these genes was assessed by small interfering RNAs targeting only the full-length or both AR isoforms . Silencing of both isoforms strongly reduced PSA , TMEPA1 , and O15393 transcript levels . Inhibition of both AR isoforms did not affect P00533 and P04626 transcript levels but decreased P00533 and increased P04626 protein levels . Proliferation of 22Rv1 cells in SDM was inhibited in the absence of AR and ARDeltaCTD . A further decrease was obtained with PKI166 , an P00533 / P04626 kinase inhibitor . Overall , we showed that ARDeltaCTD is responsible for constitutive P00533 expression and P04626 repression in 22Rv1 cells and that ARDeltaCTD and tyrosine kinase receptors are necessary for sustained 22Rv1 cell growth . Combined effects of C225 and 125-iodine seed radiation on colorectal cancer cells . BACKGROUND : To characterize the effect of combined treatment of the anti-epidermal growth factor receptor ( P00533 ) monoclonal antibody C225 and 125-iodine ( 125I ) seed radiation in human colorectal cancer . METHODS : We treated LS180 cells with 125I continuous low dose rate radiation in the presence and absence of 100 nM C225 . The clonogenic capacity , cellular proliferation , cell cycle distribution , apoptosis , and molecular pathways of the cells following the treatments were analyzed in vitro . RESULTS : The sensitizer enhancement ratio of C225 was approximately 1.4 . Treatment with C225 and radiation alone produced significant inhibition of cell growth , but combination therapy produced greater inhibition than either treatment administered alone . C225 increased the radiation-induced apoptosis and the fraction of γ- P16104 foci positive cells at 48 h after treatment . The Akt phosphorylation level was lower in the cells receiving the combination treatment than in the cells treated with radiation or C225 alone . CONCLUSIONS : These findings indicate that C225 sensitizes LS180 cells to 125I seed radiation . Growth inhibition is mediated by inducing apoptosis and not cell cycle arrest . Additionally , we confirmed that C225 impairs DNA repair by reducing the cellular level of the P78527 and P12956 proteins . Furthermore , the inhibition of Akt signaling activation may be responsible for the C225-mediated radiosensitization . A novel approach in the treatment of cancer : targeting the epidermal growth factor receptor . The epidermal growth factor receptor ( P00533 ) autocrine pathway contributes to a number of processes important to cancer development and progression , including cell proliferation , apoptosis , angiogenesis , and metastatic spread . The critical role the P00533 plays in cancer has led to an extensive search for selective inhibitors of the P00533 signaling pathway . The results of a large body of preclinical studies and the early clinical trials thus far conducted suggest that targeting the P00533 could represent a significant contribution to cancer therapy . A variety of different approaches are currently being used to target the P00533 . The most promising strategies in clinical development include monoclonal antibodies to prevent ligand binding and small molecule inhibitors of the tyrosine kinase enzymatic activity to inhibit autophosphorylation and downstream intracellular signaling . At least five blocking monoclonal antibodies have been developed against the P00533 . Among these , IMC-225 is a chimeric human-mouse monoclonal IgG1 antibody that has been the first anti- P00533 targeted therapy to enter clinical evaluation in cancer patients in Phase II and III studies , alone or in combination with conventional therapies , such as radiotherapy and chemotherapy . A number of small molecule inhibitors of the P00533 tyrosine kinase enzymatic activity is also in development . DB00530 and ZD1839 ( DB00317 ) are currently in Phase II and III development , respectively . ZD1839 , a p.o. active , selective quinazoline derivative has demonstrated promising in vitro and in vivo antitumor activity . Preliminary results from Phase I and II trials in patients with advanced disease demonstrate that ZD1839 and DB00530 have an acceptable tolerability profile and promising clinical efficacy in patients with a variety of tumor types . This mini-review describes the P00533 inhibitors in clinical development . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . Beyond the TRIBUTE trial : integrating P00533 / P00533 tyrosine kinase inhibitors with chemotherapy in advanced NSCLC . Evaluation of : Herbst RS , Prager D , Hermann R et al. : TRIBUTE : a Phase III trial of erlotinib hydrochloride ( DB00530 ) combined with carboplatin and paclitaxel chemotherapy in advanced non-small cell lung cancer . J . Clin . Oncol . 23 , 5892-5899 ( 2005 ) . Patients diagnosed with advanced non-small cell lung cancer have limited therapeutic options . A large , randomized , controlled trial of the human epidermal growth factor receptor tyrosine-kinase inhibitor , erlotinib ( Tarceva ) , plus standard first-line chemotherapy did not meet its primary end point of improved survival in the overall population , but did reveal a striking survival benefit in a subset of patients who had never smoked . There are a number of possible explanations for the lack of overall benefit , including the use of an unselected patient population , the need for alternatives to concurrent administration , and a postulated pathophysiological interaction between erlotinib and chemotherapy . Ongoing studies investigating alternative schedules and sequences of administration with chemotherapy will help clinicians to determine how agents such as erlotinib can best be combined with standard cytotoxic agents . Targeting the P00533 pathway for cancer therapy . Clinical studies have shown that HER-2/Neu is over-expressed in up to one-third of patients with a variety of cancers , including B-cell acute lymphoblastic leukemia ( B-ALL ) , breast cancer and lung cancer , and that these patients are frequently resistant to conventional chemo-therapies . Additionally , in most patients with multiple myeloma , the malignant cells over-express a number of epidermal growth factor receptors ( P00533 )s and their ligands , HB- P01133 and amphiregulin , thus this growth-factor family may be an important aspect in the patho-biology of this disease . These and other , related findings have provided the rationale for the targeting of the components of the P00533 signaling pathways for cancer therapy . Below we discuss various aspects of P00533 -targeted therapies mainly in hematologic malignancies , lung cancer and breast cancer . Beside novel therapeutic approaches , we also discuss specific side effects associated with the therapeutic inhibition of components of the P00533 -pathways . Alongside small inhibitors , such as DB01259 ( DB01259 , GW572016 ) , Gefitinib ( DB00317 , ZD1839 ) , and Erlotinib ( Tarceva , DB00530 ) , a significant part of the review is also dedicated to therapeutic antibodies ( e.g. : DB00072 /Herceptin , DB06366 / DB06366 /rhuMab-2C4 , Cetuximab/Erbitux/IMC-C225 , DB01269 /Abenix/ DB01269 , and also DB05294 ) . In addition , we summarize , both current therapy development driven by antibody-based targeting of the P00533 -dependent signaling pathways , and furthermore , we provide a background on the history and the development of therapeutic antibodies . Erlotinib in non-small cell lung cancer : a review . Erlotinib ( Tarceva , DB00530 ; Pfizer , Inc. ) is an orally-active , targeted inhibitor of the epidermal growth factor receptor ( P00533 / P00533 ) , which is part of a key regulatory pathway in cancer . Patients with advanced , incurable non-small cell lung cancer ( NSCLC ) may derive a clinical benefit from first- and second-line chemotherapy , but third-line treatment with available cytotoxic agents is not effective . Remarkably , P00533 / P00533 antagonists have demonstrated activity as second- and even third-line treatment for this disease . Erlotinib is the first of this novel class of drug to demonstrate a statistically significant and clinically relevant difference in overall survival , progression free survival and time to disease related symptoms ( cough , pain , shortness of breath ) compared with treatment with best supportive care in patients who have failed standard first- or second-line chemotherapy . This paper reviews the pharmacology , preclinical and clinical data to support the use of erlotinib in NSCLC .

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What future for combination therapies ? For most patients who require lipid-lowering treatment , statin monotherapy is the appropriate treatment . However , in those patients where statin monotherapy does not produce optimal lipid levels , the combination of a statin with niacin , a bile acid sequestrant , a fibric acid derivative , a cholesterol absorption inhibitor or a fish oil preparation may provide improved control . The choice of combination therapy depends upon the patient 's lipid profile and tolerability of the medication . Combination of a statin with niacin , a bile acid sequestrant or ezetimibe , a cholesterol absorption inhibitor , should be considered for patients with very high low-density lipoprotein cholesterol ( LDL-C ) levels , while combination with either a fibric acid derivative or a fish oil should be considered for patients with high LDL-C and high triglyceride levels . A number of new lipid-lowering agents are currently in development , including cholesteryl ester transfer protein ( P11597 ) inhibitors , acyl coenzyme A : cholesterol acyltransferase ( ACAT ) inhibitors , ileal bile acid transport ( Q12908 ) inhibitors , microsomal triglyceride transfer protein ( P55157 ) inhibitors and dual peroxisome proliferator-activated receptor ( Q07869 ) alpha and gamma agonists . Introduction of these novel therapies will provide opportunities for developing different combination strategies that may help to optimise lipid profiles in patients who are currently difficult to treat . The introduction of new combinations will require careful study to ensure that the risks of drug interactions and adverse events are minimised . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . Rat peroxisome proliferator-activated receptors and brown adipose tissue function during cold acclimatization . Brown adipose tissue ( Q14032 ) hyperplasia is a fundamental physiological response to cold ; it involves an acute phase of mitotic cell growth followed by a prolonged differentiation phase . Peroxisome proliferator-activated receptors ( PPARs ) are key regulators of fatty acid metabolism and adipocyte differentiation and may therefore mediate important metabolic changes during non-shivering thermogenesis . In the present study we have investigated Q07869 mRNA expression in relation to peroxisome proliferation in rat Q14032 during cold acclimatization . By immunoelectron microscopy we show that the number of peroxisomes per cytoplasmic volume and acyl- DB01992 oxidase immunolabeling density remained constant ( thus increasing in parallel with tissue mass and cell number ) during the initial proliferative phase and the acute thermogenic response but increased after 14 days of cold exposure , correlating with terminal differentiation of Q14032 . A pronounced decrease in Q14032 PPARalpha and PPARgamma mRNA levels was found within hours of exposure to cold , which was reversed after 14 days , suggesting a role for either or both of these subtypes in the proliferation and induction of peroxisomes and peroxisomal beta-oxidation enzymes . In contrast , PPARdelta mRNA levels increased progressively during cold exposure . Transactivation assays in HIB 1B and P29320 -293 cells demonstrated an adrenergic stimulation of peroxisome proliferator response element reporter activity via Q07869 , establishing a role for these nuclear receptors in hormonal regulation of gene transcription in Q14032 . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Possible involvement of Q07869 gamma in the regulation of basal channel opening of Q99572 receptor in cultured mouse astrocytes . AIMS : Recently , we demonstrated that cultured mouse astrocytes exhibited basal channel opening of Q99572 receptor ( P2X7R ) in the absence of any exogenous ligand , but the regulatory mechanism involved was not elucidated . Since our preliminary experiments suggested possible involvement of peroxisome proliferator-activated receptor ( Q07869 ) gamma in the regulation , we examined whether Q07869 gamma regulated P2X7R basal channel opening in mouse astrocytes . MAIN METHODS : P2X7R channel opening was assessed as to the uptake of a marker dye , YO-PRO-1 ( YP ) , in the presence or absence of agonists and antagonists for Q07869 gamma under a fluorescence microscope . Expression of Q07869 gamma was evaluated by Western blotting and immunocytochemistry . KEY FINDINGS : NSAIDs such as flufenamic acid ( FFA ) and indomethacin , which are a cyclooxygenase inhibitor and a Q07869 gamma agonist , showed enhancing and inhibiting effects on YP uptake at low and high concentrations , respectively , and the enhanced uptake was abolished by periodate-oxidized DB00171 ( oxATP ) , a selective P2X7R antagonist . The Q07869 gamma agonists 15-deoxy-Delta(12,14)-prostaglandin J(2) and ciglitazone decreased the basal and FFA-enhanced YP uptake , while the antagonist GW9662 increased YP uptake , this effect being blocked by the agonists and also by oxATP . Q07869 gamma was distributed in the nucleus and cytosolic/membrane fraction of cultured mouse astrocytes . SIGNIFICANCE : These findings indicate that basal channel opening of P2X7R in mouse astrocytes is at least in part regulated by Q07869 gamma . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Beyond statins : new lipid lowering strategies to reduce cardiovascular risk . Statins are the first-line therapy in LDL- DB04540 ( LDL-C ) reduction and its clinical use has contributed to significant prevention and treatment of atherosclerotic vascular disease . Yet , a significant proportion of patients remain at high risk . Recently , a number of new therapies have been developed to further lower LDL-C . These agents may provide clinical benefit on top of statin therapy in patients with high residual risk , severe hypercholesterolemia or as an alternative for patients who are intolerant to statins . We review four novel approaches based on the inhibition of proprotein convertase subtilisin/kexin type 9 ( Q8NBP7 ) , apolipoprotein-B100 ( apoB ) , Cholesteryl ester transport protein ( P11597 ) and microsomal triglyceride transfer protein ( P55157 ) . ApoB and P55157 inhibitors ( DB05528 and DB08827 ) are indicated only for homozygous familial hypercholesterolemia patients . The results of ongoing trials with P11597 and Q8NBP7 inhibitors may warrant a wider employment in different categories of patients at high risk for cardiovascular disease . Q07869 gene variants influence progression of coronary atherosclerosis and risk of coronary artery disease . BACKGROUND : Q07869 ( PPARalpha ) regulates the expression of genes involved in lipid metabolism and inflammation , making it a candidate gene for atherosclerosis and ischemic heart disease ( IHD ) . METHODS AND RESULTS : We investigated the association between the leucine 162 to valine ( L162V ) polymorphism and a G to C transversion in intron 7 of the PPARalpha gene and progression of atherosclerosis in the DB01241 Coronary Angiography Trial ( LOCAT ) , a trial examining the effect of gemfibrozil treatment on progression of atherosclerosis after bypass surgery and on risk of IHD in the second Northwick Park Heart Study ( Q9NP85 ) , a prospective study of healthy middle-aged men in the United Kingdom . There was no association with plasma lipid concentrations in either study . Both polymorphisms influenced progression of atherosclerosis and risk of IHD . V162 allele carriers had less progression of diffuse atherosclerosis than did L162 allele homozygotes with a similar trend for focal atherosclerosis . Intron 7 C allele carriers had greater progression of atherosclerosis than did G allele homozygotes . The V162 allele attenuated the proatherosclerotic effect of the intron 7 C allele . Homozygotes for the intron 7 C allele had increased risk of IHD , an effect modulated by the L162V polymorphism CONCLUSIONS : The PPARalpha gene affects progression of atherosclerosis and risk of IHD . Absence of association with plasma lipid concentrations suggests that PPARalpha affects atherosclerotic progression directly in the vessel wall . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Regulation of angiotensin II receptors beyond the classical pathway . The DB01367 ( renin-angiotensin system ) plays a role not only in the cardiovascular system , including blood pressure regulation , but also in the central nervous system . AngII ( angiotensin II ) binds two major receptors : the AT(1) receptor ( AngII type 1 receptor ) and AT(2) receptor ( AngII type 2 receptor ) . It has been recognized that AT(2) receptor activation not only opposes AT(1) receptor actions , but also has unique effects beyond inhibitory cross-talk with AT(1) receptor signalling . Novel pathways beyond the classical actions of DB01367 , the P12821 ( angiotensin-converting enzyme ) /AngII/AT(1) receptor axis , have been highlighted : the Q9BYF1 /Ang-(1-7) [ angiotensin-(1-7) ] /Mas receptor axis as a new opposing axis against the P12821 /AngII/AT(1) receptor axis , novel AngII-receptor-interacting proteins and various AngII-receptor-activation mechanisms including dimer formation . Q6RW13 ( AT(1)-receptor-associated protein ) and Q9ULD2 ( AT(2)-receptor-interacting protein ) are well-characterized AngII-receptor-associated proteins . These proteins could regulate the functions of AngII receptors and thereby influence various pathophysiological states . Moreover , the possible cross-talk between Q07869 ( peroxisome-proliferator-activated receptor ) -γ and AngII receptor subtypes is an intriguing issue to be addressed in order to understand the roles of DB01367 in the metabolic syndrome , and interestingly some ARBs ( AT(1)-receptor blockers ) have been reported to have an AT(1)-receptor-blocking action with a partial Q07869 -γ agonistic effect . These emerging concepts concerning the regulation of AngII receptors are discussed in the present review . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Q07869 gamma ligands , rosiglitazone and pioglitazone , inhibit P09038 - and P15692 -mediated angiogenesis . OBJECTIVE : To study the effect of peroxisome proliferator-activated receptor-gamma ( Q07869 gamma ) agonists , pioglitazone and rosiglitazone , on vascular endothelial growth factor ( P15692 ) - and basic fibroblast growth factor ( P09038 ) -induced angiogenesis and on endothelial cell migration . METHODS : Chick chorioallantoic membrane ( P62158 ) model was used to evaluate the efficacy of pioglitazone and rosiglitazone on P15692 - and P09038 -induced angiogenesis . In addition , the effect of pioglitazone and rosiglitazone on endothelial cell migration was evaluated using 8 mm pore filter to a feeder layer containing vitronectin as chemoattractant . RESULTS : Pioglitazone and rosiglitazone inhibited the pro-angiogenic effects of P09038 and P15692 in the P62158 model significantly ( P < 0.001 ) to the same extent . Endothelial cell migration was also inhibited by both pioglitazone and rosiglitazone ( P < 0.001 ) . CONCLUSIONS : These results suggest that Q07869 gamma ligands , pioglitazone and rosiglitazone , in addition to their important regulatory role in adipogenesis and inflammation , possess anti-angiogenic properties . Thus , Q07869 gamma ligands may be useful in the treatment of diabetic retinopathy , macular degeneration , and other ocular disorders and may lower the risk to develop cancer in diabetic patients . Dynamic regulation of platelet-derived growth factor receptor α expression in alveolar fibroblasts during realveolarization . Although the importance of platelet-derived growth factor receptor ( P09619 ) -α signaling during normal alveogenesis is known , it is unclear whether this signaling pathway can regulate realveolarization in the adult lung . During alveolar development , P09619 -α-expressing cells induce α smooth muscle actin ( α-SMA ) and differentiate to interstitial myofibroblasts . Fibroblast growth factor ( FGF ) signaling regulates myofibroblast differentiation during alveolarization , whereas peroxisome proliferator-activated receptor ( Q07869 ) -γ activation antagonizes myofibroblast differentiation in lung fibrosis . Using left lung pneumonectomy , the roles of FGF and Q07869 -γ signaling in differentiation of myofibroblasts from P09619 -α-positive precursors during compensatory lung growth were assessed . FGF receptor ( FGFR ) signaling was inhibited by conditionally activating a soluble dominant-negative P21802 transgene . Q07869 -γ signaling was activated by administration of rosiglitazone . Changes in α-SMA and P09619 -α protein expression were assessed in P09619 -α-green fluorescent protein ( GFP ) reporter mice using immunohistochemistry , flow cytometry , and real-time PCR . Immunohistochemistry and flow cytometry demonstrated that the cell ratio and expression levels of P09619 -α-GFP changed dynamically during alveolar regeneration and that α-SMA expression was induced in a subset of P09619 -α-GFP cells . Expression of a dominant-negative P21802 and administration of rosiglitazone inhibited induction of α-SMA in P09619 -α-positive fibroblasts and formation of new septae . Changes in gene expression of epithelial and mesenchymal signaling molecules were assessed after left lobe pneumonectomy , and results demonstrated that inhibition of P21802 signaling and increase in Q07869 -γ signaling altered the expression of Shh , FGF , Wnt , and Bmp4 , genes that are also important for epithelial-mesenchymal crosstalk during early lung development . Our data demonstrate for the first time that a comparable epithelial-mesenchymal crosstalk regulates fibroblast phenotypes during alveolar septation . Peroxisome proliferator-activated receptor-gamma agonists increase vascular endothelial growth factor expression in human vascular smooth muscle cells . Vascular endothelial growth factor ( P15692 ) , expressed in a variety of mesenchymal cells including vascular smooth muscle cells ( VSMC ) , is a potent mitogen for endothelial cells , and is used clinically applied for ischemic disease of peripheral vessels . To determine whether peroxisome proliferator-activated receptor gamma ( PPARgamma ) regulates P15692 production in VSMC , we examined P15692 secretion from VSMC treated with Q07869 agonists . DB00197 increased P15692 secretion in a time- and dose-dependent manner ( 261 +/- 35 % with 25 mM of troglitazone for 24 h ) , and also increased levels of P15692 mRNA . P15692 secretion was also increased by other PPARgamma agonists , pioglitazone , LY171883 , and 15d-PGJ2 ( 224 +/- 17.1 % , 247 +/- 36.8 % and 171 +/- 7.8 % , respectively ) , but not the PPARgamma agonists bezafibrate and Wy14643 ( 85.2 +/- 1.5 % , 94.6 +/- 3.2 , respectively ) . Our findings suggest that thiazolidinediones might be useful for the therapeutic angiogenesis for ischemic artery disease . LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Positive effects of methylphenidate on hyperactivity are moderated by monoaminergic gene variants in children with autism spectrum disorders . Methylphenidate ( DB00422 ) reduces hyperactive-impulsive symptoms common in children with autism spectrum disorders ( ASDs ) , however , response and tolerability varies widely . We hypothesized monoaminergic gene variants may moderate DB00422 effects in P51689 , as in typically developing children with attention-deficit/hyperactivity disorder . Genotype data were available for 64 children with P51689 and hyperactivity who were exposed to DB00422 during a 1-week safety/tolerability lead-in phase and 58 who went on to be randomized to placebo and three doses of DB00422 during a 4-week blinded , crossover study . Outcome measures included the Clinical Global Impression-Improvement ( CGI-I ) scale and the Aberrant Behavior Checklist ( ABC-hyperactivity index ) . A total of 14 subjects discontinued the study because of DB00422 side effects . Subjects were genotyped for variants in P21728 - P21918 , P08913 , Q01959 , P31645 , P21397 and P27338 , and P21964 . Forty-nine percent of the sample met positive responder criteria . In this modest but relatively homogeneous sample , significant differences by P21728 ( P=0.006 ) , P08913 ( P < 0.02 ) , P21964 ( P < 0.04 ) , P35462 ( P < 0.05 ) , P21917 ( P < 0.05 ) , Q01959 ( P < 0.05 ) and P31645 ( P < 0.05 ) genotypes were found for responders versus non-responders . Variants in P14416 ( P < 0.001 ) and P35462 ( P < 0.04 ) were associated with tolerability in the 14 subjects who discontinued the trial . For this first DB00422 pharmacogenetic study in children with P51689 , multiple monoaminergic gene variants may help explain individual differences in DB00422 's efficacy and tolerability . Genetics of type 2 diabetes mellitus and other specific types of diabetes ; its role in treatment modalities . Type 2 diabetes mellitus ( T2DM ) is among the most challenging health issues of the 21st century and is associated with an alarming rise in the incidence . The pathophysiological processes that lead to development of T2DM are still unclear , however impairment in insulin secretion and/or action is clearly indicated . Type 2 diabetes is a polygenic disorder with multiple genes located on different chromosomes contributing to its susceptibility . Analysis of the genetic factors is further complicated by the fact that numerous environmental factors interact with genes to produce the disorder . Only a minority of cases of type 2 diabetes are caused by single gene defects and one example is maturity onset diabetes of the young ( MODY ) . Previous studies indicated that variants in genes encoding the pancreatic β-cell K+ DB00171 channel subunits Kir6.2 ( Q14654 ) and Q09428 ( Q09428 ) are associated with neonatal diabetes . Six different types of maturity onset diabetes of young ( MODY ) have been identified based on characteristic gene defect . The common Pro12Ala polymorphism in peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) gene was confirmed in several studies to be associated with type 2 diabetes as well . More recently , studies reported variants within a novel gene , Q9NQB0 , as a putative susceptibility gene for type 2 diabetes across many ethnic backgrounds around the world . MODY patients respond better to sulphonylureas and metformin , while neonatal diabetes patients with genetic mutations can be changed from insulin to oral drugs . We hereby provide a comprehensive review on the role of genetics in type 2 diabetes mellitus . Inhibition of c-kit receptor tyrosine kinase activity by DB00619 , a selective tyrosine kinase inhibitor . DB00619 ( formerly known as CGP 57148B ) is a known inhibitor of the c-abl , bcr-abl , and platelet-derived growth-factor receptor ( P09619 ) tyrosine kinases . This compound is being evaluated in clinical trials for the treatment of chronic myelogenous leukemia . We sought to extend the activity profile of DB00619 by testing its ability to inhibit the tyrosine kinase activity of c-kit , a receptor structurally similar to P09619 . We treated a c-kit expressing a human myeloid leukemia cell line , M-07e , with DB00619 before stimulation with Steel factor ( SLF ) . DB00619 inhibited c-kit autophosphorylation , activation of mitogen-activated protein ( Q96HU1 ) kinase , and activation of Akt without altering total protein levels of c-kit , Q96HU1 kinase , or Akt . The concentration that produced 50 % inhibition for these effects was approximately 100 nmol/L . DB00619 also significantly decreased SLF-dependent growth of M-07e cells in a dose-dependent manner and blocked the antiapoptotic activity of SLF . In contrast , the compound had no effect on Q96HU1 kinase activation or cellular proliferation in response to granulocyte-macrophage colony-stimulating factor . We also tested the activity of DB00619 in a human mast cell leukemia cell line ( HMC-1 ) , which has an activated mutant form of c-kit . DB00619 had a more potent inhibitory effect on the kinase activity of this mutant receptor than it did on ligand-dependent activation of the wild-type receptor . These findings show that DB00619 selectively inhibits c-kit tyrosine kinase activity and downstream activation of target proteins involved in cellular proliferation and survival . This compound may be useful in treating cancers associated with increased c-kit kinase activity .

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Blockade of P30556 reduces oxidative stress in adipose tissue and ameliorates adipocytokine dysregulation . Dysregulated production of adipocytokines may be involved in the development of atherosclerotic cardiovascular disease in metabolic syndrome and chronic kidney disease ( CKD ) associated with metabolic syndrome . The aim of this study was to determine the effects of treatment with angiotensin II ( Ang II ) type-1 receptor blocker ( ARB ) on the regulation of adipocytokines . DB00275 , an ARB , significantly blunted the age- and body weight-associated falls in plasma adiponectin both in genetically and diet-induced obese mice , without affecting body weight , but had no effect on plasma adiponectin levels in lean mice . DB00275 also ameliorated dysregulation of adipocytokines in obesity , such as tumor necrosis factor-alpha , plasminogen activator inhibitor-1 , monocyte chemotactic protein-1 , and serum amyloid A3 . DB00275 significantly reduced reactive oxygen species originating from accumulated fat and attenuated the expression of nicotinamide adenine dinucleotide phospho hydrogenase oxidase subunits in adipose tissue . In cultured adipocytes , olmesartan acted as an antioxidant and improved adipocytokine dysregulation . Our results indicate that blockade of Ang II receptor ameliorates adipocytokine dysregulation and that such action is mediated , at least in part , by targeting oxidative stress in obese adipose tissue . Ang II signaling and subsequent oxidative stress in adipose tissue may be potential targets for the prevention of atherosclerotic cardiovascular disease in metabolic syndrome and also in metabolic syndrome-based CKD . [ Association and interaction of AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 and P35354 genes on the risk of hypertension in Antioquian population ] . INTRODUCTION : Hypertension is a multifactorial disease influenced by genetic and environmental components , with its prevalence varying across ethnic groups . Manifold studies on blood pressure regulatory system genes have been carried out -such as the renin-angiotensin-aldosterone system , the sympathetic nervous system , endothelial factor , and sodium balance- , but the results yielded were inconsistent among populations . OBJECTIVES : To evaluate the effect of both variants in genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 P35354 , and the result of the individual ancestry component on hypertension and blood pressure levels among population in Antioquia . METHODS AND MATERIALS : 107 cases and 253 controls were genotyped for 12 variants on genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 y P35354 , and for 20 ancestry informative markers . The association of polymorphisms and their interactions , and the association of ancestral genetic composition with hypertension and blood pressure levels were examined . RESULTS : Genes P35612 , rs4852706 ( OR=3.0 ; p=0.023 ) ; P21728 , rs686 ( OR=0.38 ; p=0.012 ) and P07550 , rs1042718 ( OR=10.0 ; p=0.008 ) ; as well as genotypic combinations of P21728 and P30556 ; AGT and P35611 ; and P35611 to P20020 and P35354 were associated to hypertension . The Amerindian ancestry component was associated to some decrease in diastolic blood pressure . CONCLUSION : Variants on genes P35612 , P21728 , P07550 , P30556 , AGT , P35611 , P20020 and P35354 individually or interacting , are associated to hypertension . The Amerindian ancestry component has an effect on blood pressure . Anomalous altered expressions of downstream gene-targets in P04637 -miRNA pathways in head and neck cancer . The prevalence of head and neck squamous cell carcinoma , HNSCC , continues to grow . Change in the expression of P04637 in HNSCC affects its downstream miRNAs and their gene targets , anomalously altering the expressions of the five genes , O00470 , P30556 , Q9NZJ0 , P04818 and Q16611 . These expression alterations follow the repression of P04637 that upregulates miRNA-107 , miRNA- 215 , miRNA-34 b/c and miRNA-125b , but downregulates miRNA-155 . The above five so far unreported genes are the targets of these miRNAs . Meta-analyses of microarray and RNA-Seq data followed by qRT-PCR validation unravel these new ones in HNSCC . The regulatory roles of P04637 on miRNA-155 and miRNA-125b differentiate the expressions of P30556 and BAK1in HNSCC vis-à-vis other carcinogenesis . Expression changes alter cell cycle regulation , angiogenic and blood cell formation , and apoptotic modes in affliction . Pathway analyses establish the resulting systems-level functional and mechanistic insights into the etiology of HNSCC . Angiotensin II regulates vascular and endothelial dysfunction : recent topics of P30556 signaling in the vasculature . Accumulating evidence strongly implicates angiotensin II ( AngII ) intracellular signaling in mediating cardiovascular diseases such as hypertension , atherosclerosis and restenosis after vascular injury . In vascular smooth muscle cells ( VSMCs ) , through its G-protein-coupled AngII Type 1 receptor ( AT(1) ) , AngII activates various intracellular protein kinases , such as receptor or non-receptor tyrosine kinases , which includes epidermal growth factor receptor ( P00533 ) , platelet-derived growth factor receptor ( P09619 ) , c-Src , Q14289 , Q05397 , O60674 . In addition , AngII activates serine/threonine kinases such as mitogen-activated protein kinase ( MAPK ) family , P08133 S6 kinase , Akt/protein kinase B and various protein kinase C isoforms . In VSMCs , AngII also induces the generation of intracellular reactive oxygen species ( ROS ) , which play critical roles in activation and modulation of above signal transduction . Less is known about endothelial cell ( EC ) AngII signaling than VSMCs , however , recent studies suggest that endothelial AngII signaling negatively regulates the nitric oxide ( NO ) signaling pathway and thereby induces endothelial dysfunction . Moreover , in both VSMCs and ECs , AngII signaling cross-talk with insulin signaling might be involved in insulin resistance , an important risk factor in the development of cardiovascular diseases . In fact , clinical and pharmacological studies showed that AngII infusion induces insulin resistance and AngII converting enzyme inhibitors and AT(1) receptor blockers improve insulin sensitivity . In this review , we focus on the recent findings that suggest the existence of novel signaling mechanisms whereby AngII mediates processes , such as activation of receptor or non-receptor tyrosine kinases and ROS , as well as cross-talk between insulin and NO signal transduction in VSMCs and ECs . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . Single-cell transcription site activation predicts chemotherapy response in human colorectal tumors . Candidate gene and pathway approaches , and unbiased gene expression profiling , have identified marker signatures predictive of tumor phenotypes , such as drug sensitivity and invasive or metastatic potential . However , application of such information to evaluation of tumors in the clinic is limited by cell heterogeneity in the tumor . We have developed a novel method of fluorescence in situ hybridization ( Q5TCZ1 ) that can detect transcriptional activation of individual genes at their site in single cells in the interphase nucleus . A major obstacle in the treatment of colorectal cancer is relative insensitivity to the chemotherapeutic agent DB00544 ( DB00544 ) . Here , we have developed a sensitive approach to predict relative sensitivity of colorectal cancer cells to DB00544 , using Q5TCZ1 with probes targeted to nascent mRNAs to measure the number of individual cells with active transcription sites for a panel of candidate genes . These results reveal that the transcriptional status of four key genes , thymidylate synthase ( P04818 ) , Q8WYB5 -related gene X ( Q15014 ) , Bcl2-antagonist/killer ( Q16611 ) , and ATPase , Cu(2+) transporting beta polypeptide ( P35670 ) , can accurately predict response to DB00544 . As proof of principle , we show that this transcriptional profile is predictive of response to DB00544 in a small number of patient colon tumor tissues . This approach provides a novel ability to identify and characterize unique minor cell populations in the tumor that may exhibit relative resistance to chemotherapy . Comparison of effects of olmesartan and telmisartan on blood pressure and metabolic parameters in Japanese early-stage type-2 diabetics with hypertension . P30556 blockers ( ARBs ) are regarded as first-line treatments for type-2 diabetes with hypertension . Despite the availability of various types of ARBs , there are no comparative studies of their effects on patients with diabetes . In this open-label prospective crossover study , we compared the effects of olmesartan ( 20 mg/day ) and telmisartan ( 40 mg/day ) . Twenty Japanese early-stage type-2 diabetes patients with hypertension treated with valsartan ( 80 mg/day ) for at least 8 weeks were recruited to this study . At study entry , valsartan was changed to olmesartan ( 20 mg/day ) or telmisartan ( 40 mg/day ) and administered for 8 weeks . The drugs were then switched and treatment was continued for another 8 weeks . We analyzed the blood pressure lowering effects of each drug by 24-h ambulatory blood pressure monitoring at 0 , 8 , and 16 weeks . Simultaneously , we measured metabolic parameters and inflammation markers . DB00275 lowered mean systolic and diastolic blood pressure more significantly than did telmisartan . While there were no differences between the groups in metabolic parameters , including HbA1c and adiponectin , the decreases in serum interleukin-6 and highly sensitive P02741 were more significant by olmesartan treatment . Our results indicate that olmesartan has more potent arterial blood pressure lowering and anti-inflammatory effects than telmisartan . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . A-type natriuretic peptide level in angiotensin II type 1a receptor knockout mice . A-type ( atrial ) natriuretic peptide ( P01160 ) levels in the heart and plasma were examined by immunohistochemistry , electron microscopy , and radioimmunoassay ( RIA ) in angiotensin II type 1a receptor knockout ( Agtr1a KO ) mice . Additionally , the P01160 mRNA level in the heart was measured using a real-time polymerase chain reaction ( PCR ) assay . The blood pressure in Agtr1a KO mice was significantly lower than that in wild-type ( WT ) mice . The number of P01160 granules and P01160 immunoreactivity in the auricular cardiocytes were significantly lower in Agtr1a KO mice than in WT mice . Ultrastructurally , the ventricular cardiocytes in Agtr1a KO mice occasionally had P01160 -like granules , which were not present in WT mice . The plasma , auricular , and ventricular P01160 and plasma cyclic guanosine monophosphate ( cGMP ) concentrations were significantly higher in Agtr1a KO mice than in WT mice . The P01160 mRNA levels of the auricular and ventricular cardiocytes in the Agtr1a KO mice were almost twice as large as those in WT mice . The present data suggest that a notable increase in the P01160 biosynthesis and release in the heart of Agtr1a KO mice may account for the reduction in blood pressure together with the lack of an P30556 receptor in this model . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . AT1 receptors mediate the release of prostaglandins in porcine smooth muscle cells and rat astrocytes . Angiotensin II ( AII ) can release arachidonic acid metabolites such as prostacyclin ( DB01240 ) and DB00917 from cells in cultures . It has recently been reported that the AT1 selective nonpeptide AII receptor antagonist losartan had similar effects . The present study was undertaken to further evaluate the effects of AII and losartan on cells which synthesize prostaglandins , including vascular smooth muscle , endothelial , and glial cells . Inhibition of specific [125I]AII binding was demonstrated in porcine smooth muscle cell ( PSMC ) suspensions with unlabeled AII and losartan . The IC50 values were 1.3 x 10(-9) mol/L and 7.7 x 10(-9) mol/L , respectively . PD123177 ( an P50052 selective antagonist ) had no effect on binding . AII produced a concentration-related increase in calcium mobilization ( fura-2 fluorescence ) which was blocked by losartan ( IC50 = 8.4 x 10(-8) mol/L ) but not by PD123177 ( 10(-6) mol/L ) . AII ( 10(-7) to 10(-5) mol/L ) stimulated the basal release of DB01240 by 100 % . This response was blocked by losartan ( 10(-6) to 10(-5) mol/L ) but not by PD123177 ( 10(-6) to 10(-5) mol/L ) and neither agent stimulated basal release in PSMC . Similar effects of AII and antagonists were observed upon receptor binding and DB00917 release in primary rat astrocyte ( RA ) cultures . AII did not release DB01240 from porcine endothelial cells , bovine pulmonary arterial endothelial cells , or rat P13671 glioma cells . Losartan had no significant effect at 10(-5) mol/L . By contrast , bradykinin or the calcium ionophore A23187 dramatically increased DB01240 release in each of these cells. ( ABSTRACT TRUNCATED AT 250 WORDS ) A novel role for farnesyl pyrophosphate synthase in fibroblast growth factor-mediated signal transduction . P14324 ( FPPS ) catalyses the formation of a key cellular intermediate in isoprenoid metabolic pathways . Here we describe a novel role for this enzyme in fibroblast growth factor ( FGF ) -mediated signalling . We demonstrate the binding of FPPS to FGF receptors ( FGFRs ) using the yeast two-hybrid assay , pull-down assays and co-immunoprecipitation . The interaction between FPPS and FGFR is regulated by the cellular metabolic state and by treatment with P09038 . Overexpression of FPPS inhibits P09038 -induced cell proliferation , accompanied by a failure of the P09038 -mediated induction of cyclins D1 and E. Overexpression of FPPS in fibroblasts also promotes increased farnesylation of Ras , and temporally extends P09038 -stimulated activation of the Ras/ P29323 ( extracellular-signal-regulated kinase ) cascade . These data suggest that , in addition to its role in isoprenoid biosynthesis , FPPS may function as a modulator of the cellular response to FGF treatment . Cooperative regulation of extracellular signal-regulated kinase activation and cell shape change by filamin A and beta-arrestins . beta-Arrestins ( betaarr ) are multifunctional adaptor proteins that can act as scaffolds for G protein-coupled receptor activation of mitogen-activated protein kinases ( MAPK ) . Here , we identify the actin-binding and scaffolding protein filamin A ( P21333 ) as a betaarr-binding partner using Son of sevenless recruitment system screening , a classical yeast two-hybrid system , coimmunoprecipitation analyses , and direct binding in vitro . In P21333 , the betaarr-binding site involves tandem repeat 22 in the carboxyl terminus. betaarr binds P21333 through both its N- and C-terminal domains , indicating the presence of multiple binding sites . We demonstrate that betaarr and P21333 act cooperatively to activate the MAPK extracellular signal-regulated kinase ( P29323 ) downstream of activated muscarinic M1 ( M1MR ) and angiotensin II type 1a ( P30556 ) receptors and provide experimental evidence indicating that this phenomenon is due to the facilitation of betaarr- P28482 complex formation by P21333 . In Hep2 cells , stimulation of M1MR or P30556 results in the colocalization of receptor , betaarr , P21333 , and active P29323 in membrane ruffles . Reduction of endogenous levels of betaarr or P21333 and a catalytically inactive dominant negative Q02750 , which prevents P29323 activation , inhibit membrane ruffle formation , indicating the functional requirement for betaarr , P21333 , and active P29323 in this process . Our results indicate that betaarr and P21333 cooperate to regulate P29323 activation and actin cytoskeleton reorganization . Regulation of angiotensin II type 1 receptor expression in ovarian cancer : a potential role for P38398 . BACKGROUND : Both P38398 and angiotensin II type 1 receptor ( P30556 ) play a critical role in ovarian cancer progression . However , the crosstalk between P38398 and P30556 signaling pathways remains largely unknown . METHODS : P38398 promoter methylation was analyzed by bisulfite sequence using primers focused on the core promoter region . Expression levels of P38398 and P30556 were assessed by immunohistochemistry and real-time PCR . Regression analysis was used to examine the possible relationship between P38398 and P30556 protein levels . Knockdown or overexpression of P38398 was achieved by using a lentiviral vector in 293 T cells and SKOV3 ovarian carcinoma cells , and primary non-mutated and P38398 -mutated ovarian cancer cells . RESULTS : P38398 dysfunction ( P38398 mutation or hypermethylated P38398 promoter ) ovarian cancer showed decreased P30556 levels compared to normal tissue . In contrast , P30556 expression was increased in non- P38398 -mutated ovarian cancer . Notably , P38398 activation was an effective way to induce P30556 expression in primary ovarian cancer cells and a positive correlation exists between P38398 and P30556 expression in human ovarian cancer specimens . CONCLUSIONS : These results indicate that P38398 may be a potential trigger involved in the transcriptional regulation of P30556 in the development of ovarian cancer . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Foxp3+ regulatory T cells promote lung epithelial proliferation . Acute respiratory distress syndrome ( ARDS ) causes significant morbidity and mortality each year . There is a paucity of information regarding the mechanisms necessary for ARDS resolution . Foxp3(+) regulatory T cells ( Foxp3(+) T(reg) cells ) have been shown to be an important determinant of resolution in an experimental model of lung injury . We demonstrate that intratracheal delivery of endotoxin ( lipopolysaccharide ) elicits alveolar epithelial damage from which the epithelium undergoes proliferation and repair . Epithelial proliferation coincided with an increase in Foxp3(+) T(reg) cells in the lung during the course of resolution . To dissect the role that Foxp3(+) T(reg) cells exert on epithelial proliferation , we depleted Foxp3(+) T(reg) cells , which led to decreased alveolar epithelial proliferation and delayed lung injury recovery . Furthermore , antibody-mediated blockade of CD103 , an integrin , which binds to epithelial expressed P12830 decreased Foxp3(+) T(reg) numbers and decreased rates of epithelial proliferation after injury . In a non-inflammatory model of regenerative alveologenesis , left lung pneumonectomy , we found that Foxp3(+) T(reg) cells enhanced epithelial proliferation . Moreover , Foxp3(+) T(reg) cells co-cultured with primary type II alveolar cells ( P50052 ) directly increased P50052 cell proliferation in a CD103-dependent manner . These studies provide evidence of a new and integral role for Foxp3(+) T(reg) cells in repair of the lung epithelium . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . Expression of renin-angiotensin system components in the early bovine embryo . The renin-angiotensin system ( DB01367 ) , mainly associated with the regulation of blood pressure , has been recently investigated in female reproductive organs and the developing foetus . Angiotensin II ( Ang II ) influences oviductal gamete movements and foetal development , but there is no information about DB01367 in the early embryo . The aim of this study was to determine whether DB01367 components are present in the pre-implantation embryo , to determine how early they are expressed and to investigate their putative role at this stage of development . Bovine embryos produced in vitro were used for analysis of DB01367 transcripts ( RT-PCR ) and localisation of the receptors P30556 and P50052 ( immunofluorescent labelling ) . We also investigated the effects of Ang II , DB00275 ( P30556 antagonist ) and PD123319 ( P50052 antagonist ) on oocyte cleavage , embryo expansion and hatching . Pre-implanted embryos possessed P30556 and P50052 but not the other DB01367 components . Both receptors were present in the trophectoderm and in the inner cell mass of the blastocyst . P30556 was mainly localised in granular-like structures in the cytoplasm , suggesting its internalisation into clathrin-coated vesicles , and P50052 was found mainly in the nuclear membrane and in the mitotic spindle of dividing trophoblastic cells . Treating embryos with PD123319 increased the proportion of hatched embryos compared with the control . These results , the first on DB01367 in the early embryo , suggest that the pre-implanted embryo responds to Ang II from the mother rather than from the embryo itself . This may be a route by which the maternal DB01367 influences blastocyst hatching and early embryonic development . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . P38398 , Q9BXW9 and Chk1 are potential molecular targets for the modulation of a radiation-induced DNA damage response in bystander cells . Radiotherapy is an important treatment option for many human cancers . Current research is investigating the use of molecular targeted drugs in order to improve responses to radiotherapy in various cancers . The cellular response to irradiation is driven by both direct DNA damage in the targeted cell and intercellular signalling leading to a broad range of bystander effects . This study aims to elucidate radiation-induced DNA damage response signalling in bystander cells and to identify potential molecular targets to modulate the radiation induced bystander response in a therapeutic setting . Stalled replication forks in T98G bystander cells were visualised via bromodeoxyuridine ( BrdU ) nuclear foci detection at sites of single stranded DNA . γ P16104 co-localised with these BrdU foci . P38398 and Q9BXW9 foci formed in T98G bystander cells . Using ATR mutant F02-98 hTERT and Q13315 deficient GM05849 fibroblasts it could be shown that ATR but not Q13315 was required for the recruitment of Q9BXW9 to sites of replication associated DNA damage in bystander cells whereas P38398 bystander foci were Q13315 -dependent . Phospho-Chk1 foci formation was observed in T98G bystander cells . Clonogenic survival assays showed moderate radiosensitisation of directly irradiated cells by the Chk1 inhibitor P55089 -01 but increased radioresistance of bystander cells . This study identifies P38398 , Q9BXW9 and Chk1 as potential targets for the modulation of radiation response in bystander cells . It adds to our understanding of the key molecular events propagating out-of-field effects of radiation and provides a rationale for the development of novel molecular targeted drugs for radiotherapy optimisation . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males .

MH_train_69

MH_train_69

MH_train_69

interacts_with DB00910?

multiple_choice

Examination of ceramic restorative material interfacial debonding using acoustic emission and optical coherence tomography . OBJECTIVE : CAD/ P62158 ceramic restorative material is routinely bonded to tooth substrates using adhesive cement . This study investigates micro-crack growth and damage in the ceramic/dentin adhesive interface under fatigue shear testing monitored using the acoustic emission ( AE ) technique with optical coherence tomography ( O75051 ) . METHODS : Ceramic/dentin adhesive samples were prepared to measure the shear bond strength ( SBS ) under static load . Fatigue shear testing was performed using a modified ISO14801 method . Loads in the fatigue tests were applied at 80 % , 70 % , and 60 % of the SBS to monitor interface debonding . The AE technique was used to detect micro-crack signals in static and fatigue shear bond tests . RESULTS : The results showed that the average SBS value in the static tests was 10.61±2.23MPa ( mean±standard deviation ) . The average number of fatigue cycles in which ceramic/dentin interface damage was detected in 80 % , 70 % and 60 % of the SBS were 152 , 1962 and 9646 , respectively . The acoustic behavior varied according to the applied load level . Events were emitted during 60 % and 70 % fatigue tests . A good correlation was observed between crack location in O75051 images and the number of AE signal hits . SIGNIFICANCE : The AE technique and O75051 images employed in this study could potentially be used as a pre-clinical assessment tool to determine the integrity of cemented load bearing restored ceramic material . Sustainable cyclic load stresses in ceramic/dentin-bonded specimens were substantially lower than the measured SBS . Predicted S-N curve showed that the maximum endured load was 4.18MPa passing 10(6) fatigue cyclic . Use of a cyclo-oxygenase 2 inhibitor for prophylaxis of cystoid macular oedema following cataract surgery : a randomized placebo-controlled trial . BACKGROUND : To assess the efficacy of Celecoxib , a cyclo-oxygenase 2 ( P35354 ) inhibitor , as prophylaxis for cystoid macular oedema after routine cataract surgery . METHODS : A prospective , randomized , double-blind placebo-controlled trial of 69 hospital patients undergoing cataract surgery . Celecoxib 200 mg twice daily or placebo was given immediately after surgery for 14 days . Optical coherence tomography was used to quantify macular thickness before surgery and on day 1 , week 2 and week 6 after surgery . RESULTS : Sixty-nine patients were enrolled , of which 33 received placebo and 36 received active drug . Clinically apparent cystoid macular oedema occurred in four of the treatment group and two of the placebo group ( P = 0.68 ) . No difference in best-corrected visual acuity was seen at 6 weeks ( P = 0.37 ) . Covariate analysis of the results at 2 weeks and 6 weeks showed a macular thickness of 3 % less in the treatment group compared with placebo ( P = 0.050 ) . CONCLUSION : Celecoxib may decrease macular thickening following routine cataract surgery at 2 and 6 weeks after surgery as measured by Stratus O75051 III . No difference in best-corrected visual acuity or clinically apparent cystoid macular oedema was seen . Further investigation of P35354 inhibitors in a larger prospective randomized trial is required . 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 . New analogs of vitamin D3 . Calcitriol , the most active metabolite of vitamin D , controls parathyroid gland growth and suppresses the synthesis and secretion of parathyroid hormone ( PTH ) . However , because of its potent effects on intestinal calcium absorption and bone mobilization , calcitriol treatment can induce hypercalcemia , often precluding its use at therapeutic doses . Hyperphosphatemia is also a persistent problem among patients undergoing chronic hemodialysis and can be aggravated by therapeutic doses of calcitriol . Several pharmaceutical companies were able to modify the side-chain of the 1,25(OH)2D3 , allowing some of these new analogs to retain the action on the parathyroid glands while decreasing their hypercalcemic and hyperphosphatemic effects . The structure-activity relationship for ligand-mediated transcriptional regulation has been studied in detail . In some analogs the serum binding protein ( DBP ) plays a key role in determining the pharmacokinetics of the vitamin D compound . The affinity to DBP for 22-oxacalcitriol ( O75051 ) , an analog of calcitriol for the treatment of secondary hyperparathryoidism , is approximately 300-400 times lower than that of calcitriol and the analog is rapidly cleared from the circulation . The mechanisms for the selectivity of 19-nor-1,25(OH)2D2 ( paricalcitol ) ( DB00910 ) another analog of calcitriol , is clearly different from O75051 . Although the mechanisms of action is not completely known , it does appear that paricalcitol down-regulates the P11473 in the intestine . It is likely that the unique biological profiles of vitamin D analogs in vivo are due to multiple mechanisms . Understanding the molecular basis of the analog selectivity will not only provide an explanation for their unique actions but allow intelligent design of more effective analogs in the future . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Molecular basis of a multiple lymphokine deficiency in a patient with severe combined immunodeficiency . We have previously reported that the T lymphocytes of a child with severe combined immunodeficiency are defective in the transcription of several lymphokine genes that include P60568 , P08700 , P05112 , and P05113 , which encode interleukins 2 , 3 , 4 , and 5 ( P60568 , -3 , -4 , and -5 ) . To determine whether the defect in the patient 's T lymphocytes involved a trans-acting factor common to the affected lymphokine genes , we examined the ability of nuclear factors from the patient 's T lymphocytes to bind response elements present in the regulatory region of P60568 . Nuclear factor NF-kB , activation protein 1 ( AP-1 ) , O75051 -1 , and NF-IL-2B binding activity were normal . In contrast , the binding of the nuclear factor of activated T cells ( NF-AT ) to its response element in the P60568 enhancer and to an NF-AT-like response element present in the P05112 enhancer was abnormal . To ascertain whether the abnormal NF-AT binding activity was related to an impaired function , we transfected patient and control T lymphocytes with constructs containing the reporter gene encoding chloramphenicol acetyl transferase ( CAT ) under the control of the entire P60568 regulatory region or of multimers of individual enhancer sequences . CAT expression directed by the P60568 regulatory region or by a multimer of the NF-AT-binding site was markedly lower in the patient relative to controls . In contrast , CAT gene expression directed by a multimer of the O75051 -1 proximal ( O75051 -1p ) -binding site was equivalent in patient and controls . These results indicate that an abnormality of/or influencing NF-AT may underlie the multiple lymphokine deficiency in this patient . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Vitamin D analogues . The plethora of actions attributed to 1,25(OH)2D3 throughout the body have suggested potential therapeutic applications for the treatment of hyperproliferative diseases , immune dysfunction , endocrine disorders , and metabolic bone disease . However , the potent calcemic activity of the natural vitamin D hormone has precluded its use in most cases . New vitamin D analogues are under development that display greater specificity , in most cases , by retaining the therapeutic properties of 1,25(OH)2D3 , but with lower calcemic activity . Two analogues have been approved for use in patients : calcipotriol ( DB02300 from Leo Pharmaceuticals , Copenhagen , Denmark ) for the treatment of psoriasis ; and 19-nor-1,25(OH)2D2 ( DB00910 from Abbott Laboratories , Abbott Park , IL ) for secondary hyperparathyroidism . Many others analogues are currently being tested in preclinical and clinical trials for the treatment of various types of cancer and osteoporosis , and for immunosuppression . The selectivity of the analogues can be attributed to the combined interactions with four proteins : the vitamin D receptor ( P11473 ) , the serum vitamin D binding protein ( DBP ) , the vitamin D-24-hydroxylase and to a newly described membrane receptor . Low DBP affinity has been shown to be responsible for the reduced calcemic actions of calcipotriol and 22-oxacalcitriol ( O75051 ) , which is being tested for secondary hyperparathyroidism . However , the low calcemic activity of other analogues , including 19-nor-1,25(OH)2D2 , involves other , as yet undefined , mechanisms . Understanding of the molecular basis for the selectivity of the vitamin D analogues will allow the design of more effective and safer vitamin D compounds for the treatment of a wide range of clinical disorders . The hypoxia-associated factor switches cells from HIF-1α- to HIF-2α-dependent signaling promoting stem cell characteristics , aggressive tumor growth and invasion . Most solid tumors and their metastases experience periods of low oxygen or hypoxia , which is of major clinical significance as it promotes both tumor progression and resistance to therapy . Critical mediators of the hypoxic response are the hypoxia-inducible factors HIF-1α and HIF-2α . The HIFs are nonredundant and regulate both overlapping and unique downstream target genes . Here , we describe a novel mechanism for the switch between HIF-1α- and HIF-2α-dependent transcription during tumor hypoxia caused by the hypoxia associated factor ( P00748 ) . P00748 is overexpressed in a variety of tumors and its levels are decreased during acute hypoxia , but increased following prolonged hypoxia . We have previously identified P00748 as an E3 ubiquitin ligase that binds and ubiquitinates HIF-1α by an oxygen and P40337 -independent mechanism , thus targeting HIF-1α for proteasomal degradation . Here , we show that P00748 also binds to HIF-2α , but at a different site than HIF-1α , and increases HIF-2α transactivation without causing its degradation . P00748 , thus , switches the hypoxic response of the cancer cell from HIF-1α-dependent to HIF-2α-dependent transcription and activates genes involved in invasion such as P14780 , P05121 , and the stem cell factor O75051 -3/4 . The switch to HIF-2α-dependent gene expression caused by P00748 also promotes an enriched tumor stem cell population , resulting in highly aggressive tumors in vivo . Thus , P00748 , by causing a switch from a HIF-1α- to HIF-2α-dependent response to hypoxia , provides a mechanism for more aggressive growth of tumors under prolonged hypoxia . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Association between P16109 glycoprotein ligand-1 and pathogenesis in acute coronary syndrome assessed by optical coherence tomography . OBJECTIVE : Although monocytes appear to be actively involved in the onset of acute coronary syndrome ( ACS ) , they are heterogenous in human peripheral blood . How up-regulation of monocyte subsets leads to coronary plaque rupture followed by thrombus formation remains unclear . Recent studies have shown that P16109 glycoprotein ligand-1 ( Q14242 ) is involved in monocyte activation in patients with thrombus formation . We therefore investigated the relationship between the expression of Q14242 on monocyte subsets and thrombus formation using frequency-domain optical coherence tomography ( FD- O75051 ) in patients with ACS . METHODS : We enrolled a total of 100 individuals in this study : patients with acute myocardial infarction ( AMI , n=25 ) , unstable angina pectoris ( UAP , n=20 ) , or stable angina pectoris ( n=35 ) who underwent coronary angiography , and control subjects ( n=20 ) . Three monocyte subsets ( P08571 ++CD16- , P08571 ++CD16+ , and P08571 +CD16+ ) and the expression of Q14242 were measured by flow cytometry . In patients with AMI and UAP , FD- O75051 was performed before percutaneous coronary intervention . RESULTS : Circulating peripheral P08571 ++CD16+ monocytes expressed Q14242 more frequently than P08571 ++CD16- and P08571 +CD16+ monocytes in patients with ACS . The expression of Q14242 on circulating peripheral P08571 ++CD16+ monocytes was significantly elevated in patients with AMI compared with the other 3 groups . Moreover , the expression levels of Q14242 on P08571 ++CD16+ monocytes were significantly higher in patients with plaque rupture or intracoronary thrombus assessed by FD- O75051 . CONCLUSION : Up-regulation of Q14242 on P08571 ++CD16+ monocytes may be a crucial role in plaque rupture and thrombus formation . Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . DB09301 glycosaminoglycans as major P16109 ligands on metastatic breast cancer cell lines . The metastatic breast cancer cell line , 4T1 , abundantly expresses the oligosaccharide sialylated Lewis x ( sLe(x) ) . SLe(x) oligosaccharide on tumor cells can be recognized by E- and P16109 , contributing to tumor metastatic process . We observed that both selectins reacted with this cell line . However , contrary to the P16581 reactivity , which was sLe(x) dependent , P16109 reactivity with this cell line was sLe(x)-independent . The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a) , provided a unique opportunity to characterize P16109 ligands and their contribution to metastasis in the absence of overlapping selectin ligands and P16581 binding . We observed that P16109 binding was Ca(2+)-independent and sulfation-dependent . We found that P16109 reacted primarily with cell surface chondroitin sulfate ( CS ) proteoglycans , which were abundantly and stably expressed on the surface of the 4T1 cell line . P16109 binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans ( GAGs ) . Moreover , DB01109 administration significantly inhibited experimental lung metastasis . In addition , the data suggest that surface CS GAG chains were involved in P16109 mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells . The data suggest that CS GAGs are also the major P16109 -reactive ligands on the surface of human MDA-MET cells . The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Systematic meta-analyses and field synopsis of genetic association studies in schizophrenia : the SzGene database . In an effort to pinpoint potential genetic risk factors for schizophrenia , research groups worldwide have published over 1,000 genetic association studies with largely inconsistent results . To facilitate the interpretation of these findings , we have created a regularly updated online database of all published genetic association studies for schizophrenia ( ' SzGene ' ) . For all polymorphisms having genotype data available in at least four independent case-control samples , we systematically carried out random-effects meta-analyses using allelic contrasts . Across 118 meta-analyses , a total of 24 genetic variants in 16 different genes ( P02649 , P21964 , DAO , P21728 , P14416 , P21917 , Q96EV8 , P47870 , Q13224 , HP , P01584 , P42898 , O75051 , P31645 , P04637 and P17752 ) showed nominally significant effects with average summary odds ratios of approximately 1.23 . Seven of these variants had not been previously meta-analyzed . According to recently proposed criteria for the assessment of cumulative evidence in genetic association studies , four of the significant results can be characterized as showing ' strong ' epidemiological credibility . Our project represents the first comprehensive online resource for systematically synthesized and graded evidence of genetic association studies in schizophrenia . As such , it could serve as a model for field synopses of genetic associations in other common and genetically complex disorders . Effective capillary electrophoresis-based heteroduplex analysis through optimization of surface coating and polymer networks . The efficacy of capillary electrophoresis for detecting DNA mutations via heteroduplex analysis ( HDA ) is dependent upon both the effective passivition of the capillary surface and the choice of the correct polymer network for sieving . Using HDA with laser-induced fluorescence detection of fluorescently labeled DNA fragments , an effective coating and optimal polymer matrix were sought . Optimized separation conditions were determined through the methodological evaluation of a number of different silanizing reagents , polymeric coatings , and polymer networks for resolving the PCR-amplified DNA fragments associated with five mutations ( 185delAG , 1294del40 , 4446C > G , 5382insC , 5677insA ) in the breast cancer susceptibility gene ( P38398 ) . For capillary coating , allyldimethylchlorosilane , 4-chlorobutyldimethylchlorosilane , (gamma-methacryloxypropyl)trimethoxysilane , chlorodimethyloctylsilane ( O75051 ) , and 7-octenyltrimethoxysilane were evaluated as silanizing reagents in combination with poly(vinylprrolidone) ( PVP ) and polyacrylamide ( PA ) as the polymeric coat . The HDA results were compared with those obtained using a commercial ( FC ) coated capillary . Of these , the O75051 -PVP combination was found to be most effective . Using this modified capillary , HDA with polymer networks that included DB11602 ( O14777 ) , linear polyacrylamide , and PVP showed that a PVP- , PA- , or FC-coated capillary , in combination with O14777 as the sieving polymer , could be used effectively to discriminate the mutations in less than 10 min . However , optimal performance was observed with the O75051 -PVP-coated capillary and O14777 as the polymer network . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . DB00082 increases intra-abdominal fat in patients with acromegaly : a pilot study . OBJECTIVE : Acromegalic patients have increased lipolysis and decreased fat mass as well as reduced insulin sensitivity and glucose intolerance . During somatostatin analog therapy , these changes persist despite GH suppression , but they are now due to drug-induced suppression of insulin secretion . By contrast , during pegvisomant ( PEG ) therapy , GH no longer stimulates lipolysis due to the blockade of its receptor , while insulin action is unabated . Hence , both insulin sensitivity and fat mass , including intra-abdominal fat , should increase . We therefore studied intra-abdominal fat and insulin resistance in acromegalic patients after a 3-month octreotide-washout period , i.e. , during untreated acromegaly , and during PEG treatment . METHODS : Five acromegalic patients , not controlled on octreotide ( O75051 ) therapy , were studied after 3-month O75051 washout and 6-month PEG therapy . P01308 sensitivity was determined by homeostatic model assessment value and hyperinsulinemic , normoglycemic clamp . Subcutaneous and intra-abdominal fat were measured by electron beam computed tomography . RESULTS : During PEG therapy , all the patients had normal , age-adjusted P05019 concentrations . Compared with washout , insulin sensitivity ( HOMA and M value ) was not significantly different . However , intra-abdominal fat mass increased significantly during therapy ( median ( range ) cm(2) : 112 ( 84-480 ) and 172 ( 112-524 ) respectively , P < 0.05 ) , while subcutaneous fat was not significantly different . Low-density lipoprotein cholesterol , high-density lipoprotein cholesterol , and triglycerides remained unchanged . CONCLUSIONS : During PEG therapy of acromegalic patients , intra-abdominal fat increases . Visceral obesity is a risk factor for cardiovascular disease . Hence , confirmation and further studies in a larger cohort of acromegalic patients on PEG treatment are warranted . New perspectives of vesicular monoamine transporter 2 chemical characteristics in mammals and its constant expression in type 1 diabetes rat models . Vesicular monoamine transporter 2 ( Q05940 ) has been exploited as a biomarker of β-cell mass in human islets . However , a current report suggested no immunoreactivity of Q05940 in the β cells of rat islets . To investigate the cellular localization of Q05940 in islets further , the pancreatic tissues from monkeys and humans were compared with those of rats and mice . The study was performed using among-species comparisons and a type 1 diabetes model ( T1DM ) for rats by Western blotting , double-label immunofluorescence , and confocal laser scanning microscopy . We found that Q05940 -immunoreactivity ( IR ) was distributed peripherally in the islets of rodents , but was widely scattered throughout the islets of primates . Consistent with rodent islets , Q05940 -IR did not exist in insulin ( P01308 ) -IR cells but was abundantly present in glucagon ( GLU ) -IR and pancreatic polypeptide ( PP ) -IR cells in monkey and human islets . Q05940 -IR had no colocalization with P01308 -IR in any part of the rat pancreas ( head , body , and tail ) . P01308 -IR cells were reduced dramatically in T1DM rat islets , but no significant alteration in the proportion of Q05940 -IR cells and GLU-IR cells was observed . Furthermore , a strong colocalization of Q05940 -IR with GLU-IR was distributed in the peripheral regions of diabetic islets . For the first time , the current study demonstrates the presence of Q05940 in α cells and PP cells but not in β cells in the islets of monkeys and humans . This study provides convinced morphologic evidence that Q05940 is not present in β cells . There needs to be studies for new markers for β cell mass .

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interacts_with DB03615?

multiple_choice

Exploring schizophrenia drug-gene interactions through molecular network and pathway modeling . In this study , we retrieved 39 schizophrenia-related antipsychotic drugs from the DrugBank database . These drugs had interactions with 142 targets , whose corresponding genes were defined as drug targeted genes . To explore the complexity between these drugs and their related genes in schizophrenia , we constructed a drug-target gene network . These genes were overrepresented in several pathways including : neuroactive ligand-receptor pathways , glutamate metabolism , and glycine metabolism . Through integrating the pathway information into a drug-gene network , we revealed a few bridge genes connected the sub-networks of the drug-gene network : Q12879 , O60391 , Q14957 , Q13224 , P21728 , and P14416 . These genes encode ionotropic glutamate receptors belonging to the DB01221 receptor family and dopamine receptors . DB00502 was the only drug to directly interact with these pathways and receptors and consequently may have a unique action at the drug-gene interaction level during the treatment of schizophrenia . This study represents the first systematic investigation of drug-gene interactions in psychosis . A new locus for late-onset , progressive , hereditary hearing loss DFNA20 maps to 17q25 . We report the localization of DFNA20 , a gene causing dominant , nonsyndromic , progressive hearing loss in a three-generation Midwestern family , to chromosome 17q25 . Affected family members show a bilateral , sloping , progressive , sensorineural hearing loss , first evident at 6000 and 8000 Hz , that can be identified in some family members in the early teens and is clearly evident by the early twenties . As age increases , the degree of hearing loss increases with threshold shifts seen at all frequencies . Linkage to known hereditary hearing loss loci was excluded . A genome-wide screen detected positive linkage to D17S784 ( LOD(Z) = 6.62 ; & theta ; = 0 ) . Haplotype analysis refines the DFNA20 critical region to 12 cM between D17S1806 and D17S668 . Radiation hybrid mapping with Stanford P46379 and TNG panels was used to evaluate the genes P63261 , Q14957 , Q92949 , P07237 , P09486 , and P52565 as candidates for DFNA20 . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . P07237 -mediated ER retention and proteasomal degradation of procollagen I in corneal endothelial cells . Procollagen I in corneal endothelial cells ( CECs ) is intracellularly degraded immediately after its synthesis . In this study , we investigated the mechanism of intracellular degradation of procollagen I by determining the role of protein disulfide isomerase ( P07237 ) in endoplasmic reticulum ( ER ) retention and further determined the degradation pathway of procollagen I in CECs . When association of P07237 to monomeric proalpha chains or the trimeric procollagen I carboxyl propeptides ( PICPs ) was analyzed , immune complex precipitated with anti-PICP antibody contained more P07237 than that precipitated with antibodies to monomeric chains . PICPs were completely colocalized with P07237 . When CECs were transfected with P07237 vector , procollagen I and the recombinant P07237 were colocalized in the ER , whereas CECs transfected with P07237 minus KDEL ( the ER retrieval sequence ) vector demonstrated that the two proteins were localized in the Golgi and were subsequently secreted into the medium . DB03615 ( an inhibitor of the chaperone activity of P07237 ) blocked colocalization of P07237 and procollagen I . Cells treated with chloroquine ( lysosome inhibitor ) did not alter the subcellular localization of procollagen I , because the inhibitor failed to induce the accumulation of procollagen I at Golgi . On the other hand , procollagen I was colocalized with ubiquitin in the cytoplasm , and proteasomal inhibitors further facilitated the colocalization of the two proteins and accumulation of ubiquitinated procollagen I ladders . These results suggest that association of P07237 with procollagen I , whether monomeric or trimeric , leads to ER retention of procollagen I before intracellular degradation via the ubiquitin-proteasome pathway . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . The antiangiogenic effects of a vascular endothelial growth factor decoy receptor can be monitored in vivo using contrast-enhanced ultrasound imaging . The development of antiangiogenic therapies has stimulated interest in noninvasive imaging methods to monitor response . We investigated whether the effects of a vascular endothelial growth factor decoy receptor ( DB08885 , Regeneron Pharmaceuticals , Tarrytown , NY ) could be monitored in vivo using contrast-enhanced ultrasonography ( CEUS ) . Twenty nude mice ( in two groups ) were implanted with a human melanoma cell line ( DB-1 ) . The active group received DB08885 ( 4 × 25 mg/kg over 2 weeks ) , whereas the control group received an inactive protein . An ultrasound contrast agent was injected followed by power Doppler imaging ( P07237 ) and pulse inversion harmonic imaging ( PIHI ; regular and intermittent ) . Specimens were sectioned in the same planes as the images and stained for endothelial cells ( CD31 ) , cyclooxygenase-2 ( P35354 ) , P15692 , and hypoxia ( Glut1 ) . Measures of tumor vascularity obtained with the different imaging modes were compared to immunohistochemical markers of angiogenesis . Mean tumor volume was smaller in the active group than in the control group ( 656 ± 225 vs 1,160 ± 605 mm3 ) . Overall , P07237 and P15692 correlated ( r = .34 ; p = .037 ) . Vascularity decreased from control to treated mice with intermittent PIHI , as did the expression of CD31 and P35354 ( p ≤ .02 ) , whereas P15692 increased ( p = .05 ) . CEUS appears to allow in vivo monitoring of the antiangiogenic effects of DB08885 in the DB-1 human melanoma xenograft model . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . TGBp3 triggers the unfolded protein response and P63208 -dependent programmed cell death . The Potato virus X ( PVX ) triple gene block protein 3 ( TGBp3 ) , an 8-kDa membrane binding protein , aids virus movement and induces the unfolded protein response ( UPR ) during PVX infection . TGBp3 was expressed from the Tobacco mosaic virus ( TMV ) genome ( TMV-p3 ) , and we noted the up-regulation of P63208 and several endoplasmic reticulum ( ER ) -resident chaperones , including the ER luminal binding protein ( P11021 ) , protein disulphide isomerase ( P07237 ) , calreticulin ( CRT ) and calmodulin ( P62158 ) . Local lesions were seen on leaves inoculated with TMV-p3 , but not TMV or PVX . Such lesions were the result of TGBp3-elicited programmed cell death ( P61457 ) , as shown by an increase in reactive oxygen species , DNA fragmentation and induction of P63208 expression . UPR-related gene expression occurred within 8 h of TMV-p3 inoculation and declined before the onset of P61457 . TGBp3-mediated cell death was suppressed in plants that overexpressed P11021 , indicating that UPR induction by TGBp3 is a pro-survival mechanism . Anti-apoptotic genes Bcl-xl , DB01333 -9 and Op-IAP were expressed in transgenic plants and suppressed N gene-mediated resistance to TMV , but failed to alleviate TGBp3-induced P61457 . However , TGBp3-mediated cell death was reduced in P63208 -silenced Nicotiana benthamiana plants . The combined data suggest that TGBp3 triggers the UPR and elicits P61457 in plants . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . Living patches engineered from human umbilical cord derived fibroblasts and endothelial progenitor cells . OBJECTIVE : A major shortcoming in contemporary congenital heart surgery is the lack of viable replacement materials with the capacity of growth and regeneration . Here we focused on living autologous patches engineered from human umbilical cord derived fibroblasts and endothelial progenitor cells ( EPCs ) as a ready-to-use cell source for paediatric cardiovascular tissue engineering . METHODS : EPCs were isolated from 20 ml fresh umbilical cord blood by density gradient centrifugation and myofibroblasts were harvested from umbilical cord tissue . Cells were differentiated and expanded in vitro using nutrient media containing growth factors . Before seeding , cell-phenotypes were assessed by immuno-histochemistry . Biodegradable patches fabricated from synthetic polymers ( DB00158 / P07237 ) were seeded with myofibroblasts followed by endothelialization with EPCs . All patches were cultured in a perfusion bioreactor . A subgroup of patches was additionally stimulated by cyclic strain . Analysis of the neo-tissues comprised histology , immuno-histochemistry , extracellular matrix ( Q13201 ) analysis and biomechanical testing . RESULTS : Endothelial phenotypes of EPCs before seeding were confirmed by Ac-Dil-LDL , CD 31 , von-Willebrand-Factor and P29474 staining . Histology of the seeded patches demonstrated layered viable tissue formation in all samples . The cells in the newly formed tissues expressed myofibroblast markers , such as desmin and alpha-SMA . The EPCs derived neo-endothelia showed constant endothelial phenotypes ( CD 31 , P04275 ) . major constituents of Q13201 such as collagen and proteoglycans were biochemically detected . Stress-strain properties of the patches showed features of native-analogous tissues . CONCLUSIONS : Living tissue engineered patches can be successfully generated from human umbilical cord derived myofibroblasts and EPCs . This new cell source may enable the tissue engineering of versatile , living , autologous replacement materials for congenital cardiac interventions . Thiol/disulfide exchange is a prerequisite for P61073 -tropic HIV-1 envelope-mediated T-cell fusion during viral entry . Attachment of gp120 to P01730 during HIV-1 entry triggers structural rearrangement in gp120 that enables binding to an appropriate coreceptor . Following coreceptor engagement , additional conformational changes occur in the envelope ( Env ) , resulting in fusion of virion and cell membranes . Catalysts with redox-isomerase activity , such as protein disulfide isomerase ( P07237 ) , facilitate Env conversion from its inactive to its fusion-competent conformation . We report here that anti- P07237 agents effectively block P61073 Env-mediated fusion and spread of virus infection . Exogenously added P07237 , in turn , can rescue fusion from this blockade . We further find that P07237 facilitates thiol/disulfide rearrangement in gp120 during conformational change , whereas inhibition of this redox shuffling prevents gp41 from assuming the fusogenic 6-helix bundle conformation . At the virus-cell contact site , gp120 induces assembly of P07237 , P01730 , and P61073 into a tetramolecular protein complex serving as a portal for viral entry . Our findings support the hypothesis that Env conformational change depends on a well-coordinated action of a tripartite system in which P07237 works in concert with the receptor and the coreceptor to effectively lower the activation energy barrier required for Env conformational rearrangement . Endoplasmic reticulum calcium depletion impacts chaperone secretion , innate immunity , and phagocytic uptake of cells . A number of immunological functions are ascribed to cell surface-expressed forms of the endoplasmic reticulum ( ER ) chaperone calreticulin ( CRT ) . In this study , we examined the impact of ER stress-inducing drugs upon cell surface CRT induction and the resulting immunological consequences . We showed that cell surface expression of CRT and secretion of CRT , P11021 , gp96 , and P07237 were induced by thapsigargin ( THP ) treatment , which depletes ER calcium , but not by tunicamycin treatment , which inhibits protein glycosylation . Surface expression of CRT in viable , THP-treated fibroblasts correlated with their enhanced phagocytic uptake by bone marrow-derived dendritic cells . Incubation of bone marrow-derived dendritic cells with THP-treated fibroblasts enhanced sterile P05231 production and LPS-induced generation of IL-1β , IL-12 , IL-23 , and P01375 -α . However , extracellular CRT is not required for enhanced proinflammatory responses . Furthermore , the pattern of proinflammatory cytokine induction by THP-treated cells and cell supernatants resembled that induced by THP itself and indicated that other ER chaperones present in supernatants of THP-treated cells also do not contribute to induction of the innate immune response . Thus , secretion of various ER chaperones , including CRT , is induced by ER calcium depletion . CRT , previously suggested as an eat-me signal in dead and dying cellular contexts , can also promote phagocytic uptake of cells subject to ER calcium depletion . Finally , there is a strong synergy between calcium depletion in the ER and sterile P05231 , as well as LPS-dependent IL-1β , IL-12 , IL-23 , and P01375 -α innate responses , findings that have implications for understanding inflammatory diseases that originate in the ER . Phosphodiesterase-5 inhibitor sildenafil prevents neuroinflammation , lowers beta-amyloid levels and improves cognitive performance in P05067 / P49768 transgenic mice . Memory deficit is a marker of Alzheimer 's disease ( AD ) that has been highly associated with the dysfunction of cyclic GMP ( cGMP ) signaling and an ongoing inflammatory process . Phosphodiesterase-5 ( O76074 ) inhibitors prevent the breakdown of cGMP and are currently studied as a possible target for cognitive enhancement . However , it is still unknown whether inhibition of O76074 reversed β-amyloid peptide ( Aβ ) -induced neuroinflammation in P05067 / P49768 transgenic ( Tg P05067 / P49768 ) mice . The present study evaluated the cognitive behaviors , inflammatory mediators , and cGMP/PKG/pCREB signaling in 15-month-old Tg P05067 / P49768 mice and age-matched wild-type ( WT ) mice that were treated with O76074 inhibitor sildenafil and the inhibitor of cGMP-dependent protein kinase Rp-8-Br-PET-cGMPS . In comparison with WT mice , Tg P05067 / P49768 mice were characterized by impaired cognitive ability , neuroinflammatory response , and down-regulated cGMP signaling . DB00203 reversed these memory deficits and cGMP/PKG/pCREB signaling dysfunction ; it also reduced both the soluble Aβ1-40 and Aβ1-42 levels in the hippocampus . These effects of sildenafil were prevented by intra-hippocampal infusion of the Rp-8-Br-PET-cGMPS . These results suggest that sildenafil could restore cognitive deficits in Tg P05067 / P49768 mice by the regulation of PKG/pCREB signaling , anti-inflammatory response and reduction of Aβ levels . Disulfide-dependent protein folding is linked to operation of the vitamin K cycle in the endoplasmic reticulum . A protein disulfide isomerase- Q9BQB6 redox enzyme complex appears to be responsible for vitamin P04264 2,3-epoxide reduction . Gamma-carboxylation of vitamin K-dependent proteins is dependent on formation of reduced vitamin P04264 ( Vit.K1H2 ) in the endoplasmic reticulum ( ER ) , where it works as an essential cofactor for gamma-carboxylase in post-translational gamma-carboxylation of vitamin K-dependent proteins . Vit.K1H2 is produced by the warfarin-sensitive enzyme vitamin K 2,3-epoxide reductase ( Q9BQB6 ) of the vitamin K cycle that has been shown to harbor a thioredoxin-like CXXC center involved in reduction of vitamin P04264 2,3-epoxide ( Vit.K > O ) . However , the cellular system providing electrons to the center is unknown . Here data are presented that demonstrate that reduction is linked to dithiol-dependent oxidative folding of proteins in the ER by protein disulfide isomerase ( P07237 ) . Oxidative folding of reduced RNase is shown to trigger reduction of Vit.K > O and gamma-carboxylation of the synthetic gamma-carboxylase peptide substrate FLEEL . In liver microsomes , reduced RNase-triggered gamma-carboxylation is inhibited by the P07237 inhibitor bacitracin and also by small interfering RNA silencing of P07237 in P29320 293 cells . Immunoprecipitation and two-dimensional SDS-PAGE of microsomal membrane proteins demonstrate the existence of a Q9BQB6 enzyme complex where P07237 and Q9BQB6 appear to be tightly associated subunits . We propose that the P07237 subunit of the complex provides electrons for reduction of the thioredoxin-like CXXC center in Q9BQB6 . We can conclude that the energy required for gamma-carboxylation of proteins is provided by dithiol-dependent oxidative protein folding in the ER and thus is linked to de novo protein synthesis . DB03615 inhibits the chaperone activity of protein disulfide isomerase . In the process of screening of proteins binding to ribostamycin in bovine liver using the affinity column chromatography , we found that ribostamycin inhibited the chaperone activity of protein disulfide isomerase ( P07237 ) , but it did not inhibit the isomerase activity . P07237 was identified by SDS-PAGE , Western blotting , and N-terminal amino acid sequence analysis . A 100:1 molar ratio of ribostamycin to P07237 was almost sufficient to completely inhibit the chaperone activity of P07237 . The binding affinity of ribostamycin to purified bovine P07237 was determined by the Biacore system , which gave a K(D) value of 3.19 x 10(-4) M . This suggests that ribostamycin binds to region distinct from the CGHC motif of P07237 . This is the first report to describe the inhibitor of the chaperone activity of P07237 . Antenatal maternally-administered phosphodiesterase type 5 inhibitors normalize P29474 expression in the fetal lamb model of congenital diaphragmatic hernia . PURPOSE : Pulmonary hypertension ( pHTN ) , a main determinant of survival in congenital diaphragmatic hernia ( Q8NE62 ) , results from in utero vascular remodeling . Phosphodiesterase type 5 ( O76074 ) inhibitors have never been used antenatally to treat pHTN . The purpose of this study is to determine if antenatal O76074 inhibitors can prevent pHTN in the fetal lamb model of Q8NE62 . METHODS : Q8NE62 was created in pregnant ewes . Postoperatively , pregnant ewes received oral placebo or tadalafil , a O76074 inhibitor , until delivery . Near term gestation , lambs underwent resuscitations , and lung tissue was snap frozen for protein analysis . RESULTS : Mean cGMP levels were 0.53±0.11 in placebo-treated fetal lambs and 1.73±0.21 in tadalafil-treated fetal lambs ( p=0.002 ) . Normalized expression of P29474 was 82 % ±12 % in Normal-Placebo , 61 % ±5 % in Q8NE62 -Placebo , 116 % ±6 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . Normalized expression of β-sGC was 105 % ±15 % in Normal-Placebo , 82 % ±3 % in Q8NE62 -Placebo , 158 % ±16 % in Normal- DB00820 , and 86 % ±8 % in Q8NE62 - DB00820 lambs . P29474 and β-sGC were significantly decreased in Q8NE62 ( p=0.0007 and 0.01 for P29474 and β-sGC , respectively ) , and tadalafil significantly increased P29474 expression ( p=0.0002 ) . CONCLUSIONS : O76074 inhibitors can cross the placental barrier . β-sGC and P29474 are downregulated in fetal lambs with Q8NE62 . Antenatal O76074 inhibitors normalize P29474 and may prevent in utero vascular remodeling in Q8NE62 . Echoscintigraphy : a new imaging modality for the reduction of color blooming and acoustic shadowing in contrast sonography . The purpose of this study was to develop and evaluate a new imaging modality ( echoscintigraphy ) to reduce color blooming and acoustic shadowing in contrast sonography . After injection of various amounts ( 700 to 40,000 bubbles/mL ) of the echo contrast agent SH-U 563A into a flow phantom , artificial vessels were insonated in the intermittent harmonic-power Doppler imaging ( H- P07237 ) mode . The receive gain was varied from 50 % to 75 % . The cross-sectional area ( Q13216 ) of the tube was assessed using a new summation algorithm ( echoscintigraphy ) and a conventional single-frame analysis ( S-FA ) of the H- P07237 -signals . Echoscintigraphy is based on the recording and summation of low-intensity signals that are emitted during the ultrasound ( US ) -induced destruction of microbubbles . Application of the summation algorithm at low-contrast concentration allowed a gain-independent automatic calculation of the Q13216 at medium and high gain settings . Using the S-FA method , the assessment of the vessel diameter and the Q13216 was gain-dependent and allowed correct measurements only from 60 % to 65 % gain . At a high receive-gain and high contrast concentration , S-FA resulted in an overestimation of the Q13216 up to 35.5 % . Echoscintigraphy allows correct display of contrast-filled vessels over a wide range of gain settings at low contrast concentrations , where S-FA does not adequately display echo contrast . Thus , echoscintigraphy minimizes artefacts resulting from color blooming and acoustic shadowing . Transient knockdown of presenilin-1 provokes endoplasmic reticulum stress related formation of autophagosomes in HepG2 cells . The involvement of presenilins in the endoplasmic reticulum ( ER ) related autophagy was investigated by their transient knockdown in HepG2 cells . The silencing of P49768 but not of P49810 led to cell growth impairment and decreased viability . P49768 silencing resulted in ER stress response as evidenced by the elevated levels of glucose regulated protein 78 ( Grp78 ) , protein disulfide isomerase ( P07237 ) , and P35638 ( P35638 ) and by the activation of activating transcription factor 6 ( P18850 ) . The activation of autophagy was indicated by the increased procession of microtubule-associated light chain 3 protein isoform B ( LC3B ) and by decreased phosphorylation of mammalian target of rapamycin ( P42345 ) and 70kDa ribosomal protein S6 kinase ( p70S6K ) . Formation of ER-related cytoplasmic vacuolization colocalizing with the autophagic marker LC3B was also observed . The morphological effects and LC3B activation in presenilin-1 knockdown cells could be prevented by using the phosphoinositide 3-kinase ( PI3K ) inhibitor wortmannin or by calcium chelation . The results show that presenilin-1 hampers the ER stress dependent initiation of macroautophagy . Leishmania major protein disulfide isomerase as a drug target : enzymatic and functional characterization . Leishmaniasis is a major health problem worldwide and tools available for their control are limited . Effective vaccines are still lacking , drugs are toxic and expensive , and parasites develop resistance to chemotherapy . In this context , new antimicrobials are urgently needed to control the disease in both human and animal . Here , we report the enzymatic and functional characterization of a Leishmania virulence factor , Leishmania major Protein disulfide isomerase ( LmPDI ) that could constitute a potential drug target . LmPDI possesses domain structure organization similar to other P07237 family members ( a , a ' , b , b ' and c domains ) , and it displays the three enzymatic and functional activities specific of P07237 family members : isomerase , reductase and chaperone . These results suggest that LmPDI plays a key role in assisting Leishmania protein folding via its capacity to catalyze formation , breakage , and rearrangement of disulfide bonds in nascent polypeptides . Moreover , DB00626 , a reductase activity inhibitor , and DB03615 , a chaperone activity inhibitor , were tested in LmPDI enzymatic assays and versus Leishmania promastigote in vitro cultures and Leishmania amastigote multiplication inside infected THP-1-derived macrophages . DB00626 inhibited both isomerase and reductase activities , while DB03615 had no effect on the chaperone activity . Interestingly , DB00626 blocked in vitro promastigote growth as well as amastigote multiplication inside macrophages with EC(50) values of 39 μM . These results suggest that LmPDI may constitute an interesting target for the development of new anti-Leishmania drugs .

MH_train_71

MH_train_71

MH_train_71

interacts_with DB01185?

multiple_choice

P10275 promotes the migration and invasion of upper urinary tract urothelial carcinoma cells through the upregulation of P14780 and P35354 . Dysregulated androgen receptor ( AR ) signaling is implicated in several types of tumor , including carcinomas of the prostate , breast , liver and bladder . However , the contribution of AR to the progression of upper urinary tract urothelial carcinomas ( UUTUC ) has not been fully investigated . In the present study , we demonstrated that the AR is involved in the metastasis and invasiveness of UUTUC cells . We investigated the role of the AR in UUTUC by using UUTUC-derived BFTC 909 cells . The overexpression of AR promotes the migration and invasion of BFTC 909 cells . Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses . The results showed that AR-enhanced migration and invasion of UUTUC cells are linked to the upregulation of the matrix-degrading enzyme P14780 and cyclooxygenase ( P36551 ) -2 . Subsequently , the blocking of P14780 and P35354 signaling by inhibitors suppressed AR-enhanced cell migration and invasion . The results of the present study provide evidence for the first time of the role of AR in the motility and invasion of UUT cancer cells and support the hypothesis that the AR may play a critical role in the establishment of the invasive phenotype in urothelial neoplasia of UUT . Thus , the AR may also serve as a novel biomarker and potential therapeutic target for UUT cancer . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors . P10275 targets NFkappaB and TSP1 to suppress prostate tumor growth in vivo . The androgen role in the maintenance of prostate epithelium is subject to conflicting opinions . While androgen ablation drives the regression of normal and cancerous prostate , testosterone may cause both proliferation and apoptosis . Several investigators note decreased proliferation and stronger response to chemotherapy of the prostate cancer cells stably expressing androgen receptor ( AR ) , however no mechanistic explanation was offered . In this paper we demonstrate in vivo anti-tumor effect of the AR on prostate cancer growth and identify its molecular mediators . We analyzed the effect of AR on the tumorigenicity of prostate cancer cells . Unexpectedly , the AR-expressing cells formed tumors in male mice at a much lower rate than the AR-negative controls . Moreover , the AR-expressing tumors showed decreased vascularity and massive apoptosis . AR expression lowered the angiogenic potential of cancer cells , by increasing secretion of an anti-angiogenic protein , thrombospondin-1 . AR activation caused a decrease in RelA , a subunit of the pro-survival transcription factor NFkappaB , reduced its nuclear localization and transcriptional activity . This , in turn , diminished the expression of its anti-apoptotic targets , Bcl-2 and P05231 . Increased apoptosis within AR-expressing tumors was likely due to the NFkappaB suppression , since it was restricted to the cells lacking nuclear ( active ) NFkappaB . Thus we for the first time identified combined decrease of NFkappaB and increased TSP1 as molecular events underlying the AR anti-tumor activity in vivo . Our data indicate that intermittent androgen ablation is preferable to continuous withdrawal , a standard treatment for early-stage prostate cancer . ( c ) 2007 Wiley-Liss , Inc . P10275 YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) -mediated transcription of vascular endothelial growth factor ( P15692 ) and observed altered CBP-AR binding and P15692 reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 , consistent with a role for P15692 as a neurotrophic/survival factor in motor neuron disease . P10275 (CAG)n polymorphism and androgen levels in women with systemic lupus erythematosus and healthy controls . Systemic lupus erythematosus ( SLE ) is an autoimmune disorder that affects mainly females . Therefore , interrelations between the reproductive and immune system have been assumed . Considering the complex influence of hormones and receptors , we aimed to investigate the influence of androgens and androgen receptor ( AR ) polymorphism in women with SLE . One hundred and sixteen patients and 44 healthy women were investigated . DB00624 , sex hormone-binding globulin ( P04278 ) , dehydroepiandrosterone-sulphate ( DHEAS ) concentrations and AR (CAG)n polymorphism were determined . SLE patients had significantly lower levels of total and free testosterone and DHEAS in comparison with the controls . No differences in the CAG repeat length between the groups were established . Women with two alleles carrying more than 22 CAG repeats had significantly higher levels of P04278 ( 101.51 ± 61.81 vs. 69.22 ± 45.93 nmol/l , p = 0.015 ) and DHEAS ( 3.11 ± 2.65 vs. 2.11 ± 3.06 μmol/l , p = 0.007 ) and a tendency to higher testosterone concentrations ( 2.35 ± 2.10 vs. 1.71 ± 1.70 nmol/l , p = 0.056 ) in comparison with other women . The CAG repeat length in the relatively longer (CAG)n allele was inversely related to the Systemic Lupus International Collaborating Clinics/ P10323 index ( r = -0.258 , p = 0.009 ) . In conclusion , the androgen receptor (CAG)n polymorphism is not related to the development of SLE , but it could modulate the severity of the lupus chronic damages as well as the androgen levels in women . P10275 signals regulate UDP-glucuronosyltransferases in the urinary bladder : a potential mechanism of androgen-induced bladder carcinogenesis . UDP-glucuronosyltransferases ( UGTs ) , major phase II drug metabolism enzymes , play an important role in urinary bladder cancer initiation by detoxifying carcinogens . We aimed to determine if androgens regulate P78381 expression via the androgen receptor ( AR ) pathway in the bladder . Real-time reverse transcription-polymerase chain reaction and Western blot analyses were used to assess P22309 levels in the normal urothelium SVHUC cell line stably expressed with AR and in bladder tissues from AR knockout ( ARKO ) and castrated male mice . Immunohistochemistry was also performed in radical cystectomy specimens . DB02901 ( DB02901 ) treatment in SVHUC-AR reduced mRNA expression of all the P22309 subtypes ( 19-75 % decrease ) , and hydroxyflutamide antagonized the DB02901 effects . In contrast , DB02901 showed only marginal effects on P22309 expression in SVHUC-Vector . Of note were higher expression levels of UGT1As in SVHUC-Vector than in SVHUC-AR . In ARKO mice , all the Ugt1a subtypes were up-regulated , compared to wild-type littermates . In wild-type male mice , castration increased the expression of Ugt1a8 , Ugt1a9 , and Ugt1a10 . Additionally , wild-type female mice had higher levels of Ugt1a than wild-type males . Immunohistochemical studies showed strong ( 3+ ) P22309 staining in 11/24 ( 46 % ) cancer tissues , which was significantly lower than in corresponding benign tissues [ 17/18 ( 94 % ) cases ( P = 0.0009 ) ] . These results suggest that androgen-mediated AR signals promote bladder carcinogenesis by down-regulating the expression of UGTs in the bladder . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . 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 . Targeting eIF4GI translation initiation factor affords an attractive therapeutic strategy in multiple myeloma . BACKGROUND : Deregulation of protein synthesis is integral to the malignant phenotype and translation initiation is the rate limiting stage . Therefore , eIF4F translation initiation complex components are attractive therapeutic targets . METHODS : Protein lysates of myeloma cells ( cell lines/patients ' bone marrow samples ) untreated/treated with bevacizumab were assayed for eIF4GI expression , regulation ( P15559 /proteosome dependent fragmentation ) ( WB , DB00266 , qPCR ) and targets (WB). eIF4GI was inhibited by knockdown and 4EGI-1 . Cells were tested for viability ( ELISA ) , death ( FACS ) and eIF4GI targets ( WB ) . RESULTS : Previously , we have shown that manipulation of P15692 in myeloma cells attenuated P06730 dependent translation initiation . Here we assessed the significance of eIF4GI to MM cells . We demonstrated increased expression of eIF4GI in myeloma cells and its attenuation upon P15692 inhibition attributed to elevated P15559 /proteasome dependent fragmentation and diminished mRNA levels . Knockdown of eIF4GI was deleterious to myeloma cells phenotype and expression of specific molecular targets ( Q99717 /ERα/HIF1α/c-Myc ) . Finally , we showed that the small molecule 4EGI-1 inhibits eIF4GI and causes a reduction in expression of its molecular targets in myeloma . CONCLUSION : Our findings substantiate that translation initiation of particular targets in MM is contingent on the function of eIF4GI , critical to cell phenotype , and mark it as a viable target for pharmacological intervention . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . P10275 gene mutations in androgen insensitivity syndrome cause distinct patterns of reduced activation of androgen-responsive promoter constructs . Assessment of quantitative impairment of reporter gene activation is an important strategy proving pathogenetic relevance of androgen receptor ( AR ) -gene mutations in androgen insensitivity syndrome ( AIS ) . We hypothesized the additional existence of mutation-specific patterns of reduced target gene activation . Four AR-gene mutations causing AIS , L712F , M780I , R855H , and V866M , respectively , were recreated in an AR-expression plasmid . Activation of three structurally different androgen-dependent promoters ( MMTV , (ARE)2TATA , and GRE- O75051 ) was measured in transfected CHO-cells in response to dihydrotestosterone ( DB02901 ) , testosterone , androstenedione and stanozolol ( S ) . V866M showed the lowest activity across all conditions . R855H exhibited strikingly high activation of MMTV in response to DB02901 . M780I showed markedly low activation of (ARE)2TATA by S. L712F demonstrated high activation of GRE- O75051 . In essence , each mutation was characterized in this model by a specific pattern of reduced reporter gene activation . Our AR crystal structure analyses showed that L712 and M780 may cause distinct alterations of AR-ligand- and AR-coregulator interaction interfaces supporting the experimental observations . Our data support the hypothesis that mutations of the AR-gene in AIS induce mutation-specific patterns of reduced promoter activation in vitro . Considering the diversity of natural androgen-regulated promoters , mutation-specific differences of androgen response patterns may be of relevance in vivo and consequently may influence the AIS-phenotype . Assessment of transactivation patterns in vitro may be an interesting concept to extend functional description of AR-gene mutations in AIS . Delayed haemolytic transfusion reaction caused by anti-M antibody in a patient receiving interleukin-2 and interferon for metastatic renal cell cancer . Anti-M is usually a naturally occurring cold-reactive immunoglobulin M ( IgM ) antibody , often with an immunoglobulin G ( IgG ) component , and is seldom implicated in delayed haemolytic transfusion reactions ( P10275 ) . However , cases have been reported . In the majority , a P10275 is not suspected until further blood is requested and a new antibody is detected on pretransfusion testing . We describe the case of a young man receiving therapy with interleukin-2 ( P60568 ) and interferon-alpha ( IFN-alpha ) for metastatic renal cell cancer who developed a clinically suspected P10275 that was confirmed serologically to be caused by anti-M , reactive at 37 degrees C . We discuss the possible role of his biochemotherapy in the development of the P10275 . The expression of the solute carriers Q14973 and O75051 -1 is regulated by cholesterol in HepG2 cells . Drug disposition and response are greatly determined by the activities of drug-metabolizing enzymes and transporters . While the knowledge in terms of CYP enzymes and efflux ABC transporters ( such as P08183 , P-glycoprotein ) is quite extensive , influx transporters are increasingly being unveiled as key contributors to the process of drug disposition . There is little information on the regulation of these proteins in human cells , especially as regards the effect of endogenous compounds . In this study , we analysed the expression of P08684 and three uptake transporters Q14973 ( Q14973 ) , P46721 / P46721 ( P46721 ) and O75051 -1 ( O15245 ) in HepG2 cells following treatment with cholesterol . While P08684 and P46721 expression was unaffected , cholesterol treatment led to increased levels of Q14973 and O75051 -1 mRNAs . Alterations in the functional characteristics and/or expression levels of drug transporters in the liver may conceivably contribute to the variability in drug oral bioavailability often observed in the clinical settings . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin alters postnatal development of seminal vesicle epithelium . 2,3,7,8-Tetrachlorodibenzo-p-dioxin ( TCDD ) has been shown to alter male reproductive development of laboratory animals through in utero and lactational exposure . As a result of exposure , the accessory glands of the male reproductive tract , including the seminal vesicle , are decreased in size as determined by total weight of the tissue . Analysis of seminal vesicle weights over time suggests that the changes may be transient . Administration of 1.0 microg/kg TCDD during gestation caused a significant decrease in seminal vesicle weights of offspring 8-11 months of age . We examined the effects of TCDD on seminal vesicles from rats exposed in utero and lactationally . Pregnant Long Evans rats were gavaged on gestation day 15 with 1.0 microg/kg TCDD in corn oil . Male pups were euthanized and necropsied on postnatal days ( P01160 ) 15 , 25 , 32 , 49 , 63 , and 120 . Seminal vesicles were weighed and then fixed in 10 % neutral buffered formalin and processed for microscopic examination . Seminal vesicle weights were not significantly decreased until P01160 32 . P10275 mRNA expression in P01160 25 seminal vesicles was not different from control . In the present study , TCDD exposure decreased seminal vesicle epithelial branching and differentiation . Control epithelial cells had tall columnar morphology with relatively abundant cytoplasm , whereas TCDD-treated cells had rounded nuclei and less cytoplasm . In addition , immunolocalization of proliferating nuclear antigen was confined to undifferentiated basal epithelial cells of controls but was found in both basal and luminal cells of the treated seminal vesicle . Results indicate that the TCDD-induced impaired growth of the rat seminal vesicles is associated with a dramatic decrease in the development of the epithelium . Epigenetic repression of miR-31 disrupts androgen receptor homeostasis and contributes to prostate cancer progression . P10275 signaling plays a critical role in prostate cancer pathogenesis . Yet , the regulation of androgen receptor signaling remains elusive . Even with stringent androgen deprivation therapy , androgen receptor signaling persists . Here , our data suggest that there is a complex interaction between the expression of the tumor suppressor miRNA , miR-31 , and androgen receptor signaling . We examined primary and metastatic prostate cancer and found that miR-31 expression was reduced as a result of promoter hypermethylation , and importantly , the levels of miR-31 expression were inversely correlated with the aggressiveness of the disease . As the expression of androgen receptor and miR-31 was inversely correlated in the cell lines , our study further suggested that miR-31 and androgen receptor could mutually repress each other . Upregulation of miR-31 effectively suppressed androgen receptor expression through multiple mechanisms and inhibited prostate cancer growth in vivo . Notably , we found that miR-31 targeted androgen receptor directly at a site located in the coding region , which was commonly mutated in prostate cancer . In addition , miR-31 suppressed cell-cycle regulators including Q01094 , Q14209 , Q9UQ84 , Q08050 , and MCM2 . Together , our findings suggest a novel androgen receptor regulatory mechanism mediated through miR-31 expression . The downregulation of miR-31 may disrupt cellular homeostasis and contribute to the evolution and progression of prostate cancer . We provide implications for epigenetic treatment and support clinical development of detecting miR-31 promoter methylation as a novel biomarker .

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Clathrin-dependent internalization of the angiotensin II AT₁A receptor links receptor internalization to P35354 protein expression in rat aortic vascular smooth muscle cells . The major effects of Angiotensin II ( AngII ) in vascular tissue are mediated by AngII AT1A receptor activation . Certain effects initiated by AT1A receptor activation require receptor internalization . In rat aortic vascular smooth muscle cells ( RASMC ) , AngII stimulates cyclooxygenase 2 protein expression . We have previously shown this is mediated by β-arrestin-dependent receptor internalization and NF-κB activation . In this study , a specific inhibitor of clathrin-mediated endocytosis ( CME ) , pitstop-2 , was used to test the hypothesis that clathrin-dependent internalization of activated AT1A receptor mediates NF-κB activation and subsequent cyclooxygenase 2 expression . Radioligand binding assays , real time qt-PCR and immunoblotting were used to document the effects of pitstop-2 on AngII binding and signaling in RASMC . Laser scanning confocal microscopy ( LSCM ) was used to image pitstop-2׳s effects on AT1 receptor/GFP internalization in P29320 -293 cells and p65 NF-κB nuclear localization in RASMC . Pitstop-2 significantly inhibited internalization of AT1A receptor ( 44.7 % ± 3.1 % Control vs. 13.2 % ± 8.3 % Pitstop-2 ; n=3 ) as determined by radioligand binding studies in RASMC . Studies utilizing AT1A receptor/GFP expressed in P29320 293 cells and LSCM confirmed these findings . Pitstop-2 significantly inhibited AngII-induced p65 NF-κB phosphorylation and nuclear localization , P35354 message and protein expression in RASMC without altering activation of Q8NFH3 /44 P29323 or TNFα signaling . Pitstop-2 , a specific inhibitor of clathrin-mediated endocytosis , confirms that internalization of activated AT1A receptor mediates AngII activation of cyclooxygenase 2 expression in RASMC . These data provide support for additional intracellular signaling pathways activated through β-arrestin mediated internalization of G protein-coupled receptors , such as AT1A receptors . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Nuclear factor kappa B inhibition improves conductance artery function in type 2 diabetic mice . BACKGROUND : We previously reported that enhanced nuclear factor kappa B ( NFκB ) activity is responsible for resistance arteries dysfunction in type 2 diabetic mice . METHODS : In this study , we aimed to determine whether augmented NFκB activity also impairs conductance artery ( thoracic aorta ) function in type 2 diabetic mice . We treated type 2 diabetic ( db(-) /db(-) ) and control ( db(-) /db(+) ) mice with two NFκB inhibitors ( dehydroxymethylepoxyquinomicin , 6 mg/kg , twice a week and IKK-NBD peptide , 500 µg/kg/day ) for 4 weeks . RESULTS : As expected , the NFκB inhibition did not affect blood glucose level and body weight . Thoracic aorta vascular endothelium-dependent relaxation ( EDR ) , determined by the wire myograph , was impaired in diabetic mice compared with control and was significantly improved after NFκB inhibition . Interestingly , thoracic EDR was also rescued in db(-) /db(-p50NFκB-/-) and db(-) /db(- P09874 -/-) double knockout mice compared with db(-) /db(-) mice . Similarly , the acute in vitro down regulation of NFκB-p65 using p65 shRNA lentiviral particles in arteries from db(-) /db(-) mice also improved thoracic aorta EDR . Western blot analysis showed that the p65NFκB phosphorylation , cleaved P09874 and P35354 expression were increased in thoracic aorta from diabetic mice , which were restored after NFκB inhibition and in db(-) /db(-p-50NFκB-/-) and db(-) /db(- P09874 -/-) mice . CONCLUSIONS : The present results indicate that in male type 2 diabetic mice , the augmented NFκB activity also impairs conductance artery function through P09874 and P35354 -dependent mechanisms . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . P55210 ablation modulates UPR , reprograms TRAF2-JNK apoptosis and protects T17M rhodopsin mice from severe retinal degeneration . The UPR is activated in the mouse retina expressing misfolded T17M rhodopsin ( P08100 ) during autosomal dominant retinitis pigmentosa ( Q99541 ) progression . Therefore , the goal of this study is to validate the UPR-induced caspase-7 as a new therapeutic target that modulates the UPR , reduces the level of apoptosis and protects the Q99541 retina from retinal degeneration and light-induced damage . Mice were analyzed using ERG , SD- O75051 and histology to determine the role of caspase-7 ablation . The results of these experiments demonstrate the significant preservation of photoreceptors and their function in T17M P08100 P55210 retinas from P30 to P90 compared with control mice . These mice were also protected from the light-induced decline in the ERG responses and apoptosis . The RNA and protein analyses of T17M P08100 +Csp7-siRNA , Tn+Csp7-siRNA 661W cells and T17M P08100 P55210 retinas revealed that caspase-7 ablation reprograms the UPR and reduces JNK-induced apoptosis . This reduction is believed to occur through the downregulation of the P42345 and Hif1a proteins . In addition , decline in activated P09874 was detected in T17M P08100 P55210 retina . Altogether , our findings indicate that the targeting of caspase-7 in T17M P08100 mice could be a feasible therapeutic strategy for advanced stages of Q99541 . Effect of valproic acid through regulation of DB01221 receptor- P29323 signaling in sleep deprivation rats . Although the effect of mood stabilizer valproic acid ( DB00313 ) through multiple signaling pathways has been shown , its therapeutic mechanism is still largely unknown . We investigated the effect of DB00313 ( 200 mg/kg , every 12 h ) in sleep deprivation ( SD ) rats ( 72 h ) , the manic-like animal model , focusing on the N-methyl-D : -aspartic acid ( DB01221 ) receptor and signaling mediators of synaptic plasticity such as extracellular signal-regulated protein kinase ( P29323 ) , DB02527 response element-binding protein ( CREB ) , B cell chronic lymphocytic leukemia/lymphoma 2 ( P10415 ) , and brain-derived neurotrophic factor ( P23560 ) . SD reduced the expression of the Q13224 subunit of the DB01221 receptor in the frontal cortex and hippocampus but did not affect the expression of Q9UHB4 and Q12879 subunits . In comparison , DB00313 inhibited the SD-induced reduction of Q13224 expression in both brain regions . In addition , SD attenuated P29323 phosphorylation in the frontal cortex and hippocampus , whereas DB00313 prevented the attenuation . DB00313 also protected the SD-induced decrease of CREB phosphorylation , P10415 expression , and P23560 expression in the frontal cortex but not in the hippocampus . These results indicate that DB00313 could regulate DB01221 receptor- P29323 signaling in SD rats , preventing the SD-induced decrease of the expression of Q13224 subunit and the activation of P29323 signaling mediators such as P29323 , CREB , P10415 , and P23560 . Endothelial cell-derived nitric oxide enhances aerobic glycolysis in astrocytes via HIF-1α-mediated target gene activation . Astrocytes exhibit a prominent glycolytic activity , but whether such a metabolic profile is influenced by intercellular communication is unknown . Treatment of primary cultures of mouse cortical astrocytes with the nitric oxide ( NO ) donor DetaNONOate induced a time-dependent enhancement in the expression of genes encoding various glycolytic enzymes as well as transporters for glucose and lactate . Such an effect was shown to be dependent on the hypoxia-inducible factor HIF-1α , which is stabilized and translocated to the nucleus to exert its transcriptional regulation . NO action was dependent on both the PI3K/Akt/ P42345 and MEK signaling pathways and required the activation of P36551 , but was independent of the soluble guanylate cyclase pathway . Furthermore , as a consequence of NO treatment , an enhanced lactate production and release by astrocytes was evidenced , which was prevented by downregulating HIF-1α . Several brain cell types represent possible sources of NO . It was found that endothelial cells , which express the endothelial NO synthase ( P29474 ) isoform , constitutively produced the largest amount of NO in culture . When astrocytes were cocultured with primary cultures of brain vascular endothelial cells , stabilization of HIF-1α and an enhancement in glucose transporter-1 , hexokinase-2 , and monocarboxylate transporter-4 expression as well as increased lactate production was found in astrocytes . This effect was inhibited by the NOS inhibitor l-NAME and was not seen when astrocytes were cocultured with primary cultures of cortical neurons . Our findings suggest that endothelial cell-derived NO participates to the maintenance of a high glycolytic activity in astrocytes mediated by astrocytic HIF-1α activation . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . DB04630 stimulates vascular smooth muscle cell proliferation via big mitogen-activated protein kinase 1 activation . The nongenomic effects of aldosterone have been implicated in the pathogenesis of various cardiovascular diseases . DB04630 -induced nongenomic effects are attributable in part to the activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) , a classical mitogen-activated protein ( Q96HU1 ) kinase . Q13164 ( Q13164 ) , a newly identified Q96HU1 kinase , has been shown to be involved in cell proliferation , differentiation , and survival . We examined whether aldosterone stimulates Q13164 -mediated proliferation of cultured rat aortic smooth muscle cells ( RASMCs ) . P08235 ( MR ) expression and localization were evaluated by Western blotting analysis and fluorolabeling methods . P27361 /2 and Q13164 activities were measured by Western blotting analysis with the respective phosphospecific antibodies . Cell proliferation was determined by Alamar Blue colorimetric assay . DB04630 ( 0.1 to 100 nmol/L ) dose-dependently activated Q13164 in RASMCs , with a peak at 30 minutes . To clarify whether aldosterone-induced Q13164 activation is an MR-mediated phenomenon , we examined the effect of eplerenone , a selective MR antagonist , on aldosterone-induced Q13164 activation . DB00700 ( 0.1 to 10 micromol/L ) dose-dependently inhibited aldosterone-induced Q13164 activation in RASMCs . DB04630 also stimulated RASMC proliferation , which was inhibited by eplerenone . DB04630 -mediated phenomena were concluded to be attributable to a nongenomic effect because cycloheximide failed to inhibit aldosterone-induced Q13164 activation . Transfection of dominant-negative Q96HU1 kinase/ P29323 kinase 5 ( Q13163 ) , which is an upstream regulator of Q13164 , partially inhibited aldosterone-induced RASMC proliferation , which was almost completely inhibited by MEK inhibitor PD98059 . In addition to the classical steroid activity , rapid nongenomic effects induced by aldosterone may represent an alternative etiology for vascular diseases such as hypertension . Modulation of sympathetic nerve activity by microinjection of the P08908 receptor agonist 8-OH-DPAT into the rostroventrolateral medulla . In the present study , renal sympathetic nerve activity was recorded simultaneously with sympathetic nerve activity to skeletal muscle vasculature to determine if the sympatho-inhibition evoked by microinjection of the P08908 receptor agonist 8-hydroxy-2-(di-n-propylamino)teralin ( 8-OH-DPAT ) into the rostroventrolateral medulla ( RVLM ) was uniform or regional . Three patterns of sympatho-inhibition were observed in these sympathetic outflows and the type of response depended upon location of microinjection within the subretrofacial nucleus ( P11831 ) . Inhibition of renal nerve activity only was elicited by microinjections at rostral sites at the caudal pole of the facial nucleus . In contrast , inhibition of muscle sympathetic nerve activity was evoked from more caudal injections at the rostral pole of the inferior olives . Microinjection in the area between these two regions produced inhibition of both sympathetic outflows . This study demonstrates that differential inhibition of regional sympathetic outflows can be elicited by microinjection of the P08908 receptor agonist 8-OH-DPAT into the RVLM . These data suggests that this modulation is due to differences in anatomical arrangement of the medullary neurons rather than differences in neuron sensitivity to the serotonergic agonist . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Coronavirus 3CLpro proteinase cleavage sites : possible relevance to P49591 virus pathology . BACKGROUND : Despite the passing of more than a year since the first outbreak of Severe Acute Respiratory Syndrome ( P49591 ) , efficient counter-measures are still few and many believe that reappearance of P49591 , or a similar disease caused by a coronavirus , is not unlikely . For other virus families like the picornaviruses it is known that pathology is related to proteolytic cleavage of host proteins by viral proteinases . Furthermore , several studies indicate that virus proliferation can be arrested using specific proteinase inhibitors supporting the belief that proteinases are indeed important during infection . Prompted by this , we set out to analyse and predict cleavage by the coronavirus main proteinase using computational methods . RESULTS : We retrieved sequence data on seven fully sequenced coronaviruses and identified the main 3CL proteinase cleavage sites in polyproteins using alignments . A neural network was trained to recognise the cleavage sites in the genomes obtaining a sensitivity of 87.0 % and a specificity of 99.0 % . Several proteins known to be cleaved by other viruses were submitted to prediction as well as proteins suspected relevant in coronavirus pathology . Cleavage sites were predicted in proteins such as the cystic fibrosis transmembrane conductance regulator ( P13569 ) , transcription factors CREB-RP and O75051 -1 , and components of the ubiquitin pathway . CONCLUSIONS : Our prediction method NetCorona predicts coronavirus cleavage sites with high specificity and several potential cleavage candidates were identified which might be important to elucidate coronavirus pathology . Furthermore , the method might assist in design of proteinase inhibitors for treatment of P49591 and possible future diseases caused by coronaviruses . It is made available for public use at our website : http://www.cbs.dtu.dk/services/NetCorona/ . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Celecoxib blocks cardiac Kv1.5 , Kv4.3 and Kv7.1 ( P51787 ) channels : effects on cardiac action potentials . Celecoxib is a P35354 inhibitor that has been related to an increased cardiovascular risk and that exerts several actions on different targets . The aim of this study was to analyze the effects of this drug on human cardiac voltage-gated potassium channels ( Kv ) involved on cardiac repolarization Kv1.5 ( I(Kur) ) , Kv4.3+ Q9NS61 ( I(to1) ) and Kv7.1+ P15382 ( I(Ks) ) and to compare with another P35354 inhibitor , rofecoxib . Currents were recorded in transfected mammalian cells by whole-cell patch-clamp . Celecoxib blocked all the Kv channels analyzed and rofecoxib was always less potent , except on Kv4.3+ Q9NS61 channels . Kv1.5 block increased in the voltage range of channel activation , decreasing at potentials positive to 0 mV . The drug modified the activation curve of the channels that became biphasic . Block was frequency-dependent , increasing at fastest frequencies . Celecoxib effects were not altered by DB08837 (out) in R487Y mutant Kv1.5 channels but the kinetics of block were slower and the degree of block was smaller with DB08837 (in) , indicating that celecoxib acts from the cytosolic side . We confirmed the blocking properties of celecoxib on native Kv currents from rat vascular cells , where Kv1.5 are the main contributors ( IC(50)≈ 7 μM ) . Finally , we demonstrate that celecoxib prolongs the action potential duration in mouse cardiac myocytes and shortens it in guinea pig cardiac myocytes , suggesting that Kv block induced by celecoxib may be of clinical relevance . Opposite effect of Hsp90α and Hsp90β on P29474 ability to produce nitric oxide or superoxide anion in human embryonic kidney cells . Heat shock protein 90 subfamily is composed by two cytosolic isoforms known as Hsp90α and Hsp90β . Endothelial nitric oxide synthase ( P29474 ) is regulated by Hsp90 , however the specific role of each Hsp90 isoform on NO production has not been established . This study was designed to evaluate the effect of Hsp90α and Hsp90β over-expression on P29474 /NO pathway . Rat Hsp90α and Hsp90β were cloned into pcDNA3.1(+) and transfected in human embryonic kidney cells ( P29320 -293 ) . Hsp90α and Hsp90β transfection was corroborated by Western blot analysis and their effect on NO production ( NO(2)/NO(3) ) , P29474 protein and its phosphorylation at Ser1177 and Thr495 , as well as Akt/ P31749 Ser473 phosphorylation was determined . The interaction of Hsp90α and Hsp90β with P29474 and the dimer/monomer ratio of Hsp90 , as well as O(2)(-) generation were also assessed . After transfection , Hsp90α and Hsp90β levels were significantly increased in P29320 -293 cells . The Hsp90α over-expression induced a significant increase in NO(2)/NO(3) levels , an effect that was associated with increased phosphorylation of P29474 DB00133 1177 and Akt/ P31749 Ser473 , as well as with a greater Hsp90α dimerization . Noteworthy , pcHsp90β transfection reduced significantly NO(2)/NO(3) and increased O(2)(-) generation . These effects were associated with a reduction of P29474 dimeric conformation , increased P29474 Thr495 phosphorylation , reduced Akt/ P31749 phosphorylation , and by a greater amount of monomeric Hsp90β conformation . These data show for first time that Hsp90α and Hsp90β differentially modulate NO and O(2)(-) generation by P29474 through promoting changes in P29474 conformation and phosphorylation state . Differential regulation of cystic fibrosis transmembrane conductance regulator by interferon gamma in mast cells and epithelial cells . P13569 ( P13569 ) is a P13569 in epithelial cells ; recently , we identified it in mast cells . Previous work that we confirmed showed that interferon gamma ( IFNgamma ) down-regulated P13569 expression in epithelial cells ( T84 ) , but by contrast , we found that IFNgamma up-regulated P13569 mRNA and protein expression in rat and human mast cells . IFNgamma up-regulation of P13569 in mast cells was inhibited by p38 and extracellular signal-regulated kinase ( P29323 ) kinase inhibitors but not a Janus tyrosine kinase (JAK)2 inhibitor , whereas in T84 cells IFNgamma-mediated down-regulation of P13569 was O60674 -dependent and P29323 - and p38-independent . Furthermore , IFNgamma down-regulation of P13569 in T84 epithelial cells was P42224 -dependent , but up-regulation of P13569 in mast cells was P42224 -independent . Thus , differential regulatory pathways of P13569 expression in mast cells and epithelial cells exist that depend upon either p38/ P29323 or JAK/ P35610 pathways , respectively . Surprisingly , IFNgamma treatment of mast cells inhibited Cl(-) efflux , in contrast to up-regulation of P13569 /mRNA and protein expression . However , down-regulation of Cl(-) flux correlated with IFNgamma-mediated inhibition of mediator secretion . This and other work suggests that the effect of IFNgamma on P13569 expression in mast cells is important for their function . Binding of antipsychotic drugs to human brain receptors focus on newer generation compounds . Using radioligand binding assays and post-mortem normal human brain tissue , we obtained equilibrium dissociation constants ( K(d)s ) for nine new antipsychotic drugs ( iloperidone , melperone , olanzapine , ORG 5222 , quetiapine , risperidone , sertindole , ziprasidone , and zotepine ) , one metabolite of a new drug ( 9-OH-risperidone ) , and three older antipsychotics ( clozapine , haloperidol , and pimozide ) at nine different receptors ( alpha1-adrenergic , alpha2-adrenergic , dopamine D2 , histamine H1 , muscarinic , and serotonin P08908 , P28221 , 5- Q13049 , and P28335 receptors ) . DB04946 was the most potent drug at the two adrenergic receptors . ORG 5222 was the most potent drug at dopamine D2 and 5-HT2c receptors , while ziprasidone was the most potent compound at three serotonergic receptors ( P08908 , P28221 , and 5- Q13049 ) . At the remaining two receptors , olanzapine was the most potent drug at the histamine H1 receptor ( Kd=0.087 nM ) ; clozapine at the muscarinic receptor ( Kd=9 nM ) . Certain therapeutic and adverse effects , as well as certain drug interactions can be predicted from a drug 's potency for blocking a specific receptor . These data can provide guidelines for the clinician in the choice of antipsychotic drug . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Association of P35462 and Q13224 with impulse control and related behaviors in Parkinson 's disease . We aimed to assess whether allelic variants of dopamine receptor , glutamate receptor , and serotonin transporter genes are associated with the appearance of impulse control and related behaviors ( ICRB ) in Parkinson 's disease ( PD ) with dopamine replacement therapy ( P29323 ) . We surveyed ICRB in consecutive Korean patients with PD who were treated with stable P29323 using modified Minnesota Impulsive Disorders Interview over a period of 4 months . In the 404 patients who completed the interview and the 559 Korean healthy normal controls , genotyping was performed for variants of the P35462 p.S9G , P14416 Taq1A , Q13224 c.366C > G , c.2664C > T and c.-200T > G , and the promoter region of the serotonin transporter gene ( 5-HTTLPR ) . Behavioral abnormalities suggestive of ICRB including compulsive buying , gambling , sexual behavior and eating , and punding , were present in 14.4 % of the patients . Variants of P14416 and 5-HTTLPR were not associated with the risk of developing ICRB . However , the AA genotype of P35462 p.S9G and the CC genotype of Q13224 c.366C > G were more frequent in patients with ICRB than in nonaffected patients ( odds ratio [ OR ] = 2.21 , P = 0.0094 ; and 2.14 , P = 0.0087 , after adjusting for age and sex ) . After controlling for clinical variables in the multivariate analysis , carriage of either AA genotype of P35462 or CC genotype of Q13224 was identified as an independent risk factor for ICRB ( adjusted OR : 2.57 , P = 0.0087 ) . Variants of P35462 p.S9G and Q13224 c.366C > G may be associated with the appearance of ICRB in PD . P50406 mediates defective brain development in monoamine oxidase A-deficient mouse embryos . Monoamine oxidases A and B ( P21397 and P27338 ) are enzymes of the outer mitochondrial membrane that metabolize biogenic amines . In the adult central nervous system , MAOs have important functions for neurotransmitter homeostasis . Expression of MAO isoforms has been detected in the developing embryo . However , suppression of P27338 does not induce developmental alterations . In contrast , targeted inhibition and knockdown of P21397 expression ( E7.5-E10.5 ) caused structural abnormalities in the brain . Here we explored the molecular mechanisms underlying defective brain development induced by P21397 knockdown during in vitro embryogenesis . The developmental alterations were paralleled by diminished apoptotic activity in the affected neuronal structures . Moreover , dysfunctional P21397 expression led to elevated levels of embryonic serotonin ( 5-hydroxytryptamine ( 5-HT ) ) , and we found that knockdown of serotonin receptor-6 ( 5-Htr6 ) expression or pharmacologic inhibition of 5-Htr6 activity rescued the P21397 knockdown phenotype and restored apoptotic activity in the developing brain . Our data suggest that excessive 5-Htr6 activation reduces activation of caspase-3 and -9 of the intrinsic apoptotic pathway and enhances expression of antiapoptotic proteins Bcl-2 and Bcl-XL . Moreover , we found that elevated 5-HT levels in P21397 knockdown embryos coincided with an enhanced activation of extracellular signal-regulated kinase 1/2 ( P27361 /2 ) and a reduction of proliferating cell numbers . In summary , our findings suggest that excessive 5-HT in P21397 -deficient mouse embryos triggers cellular signaling cascades via 5-Htr6 , which suppresses developmental apoptosis in the brain and thus induces developmental retardations . P42345 supports long-term self-renewal and suppresses mesoderm and endoderm activities of human embryonic stem cells . Despite the recent identification of the transcriptional regulatory circuitry involving P48431 , Q9H9S0 , and O75051 -4 , the intracellular signaling networks that control pluripotency of human embryonic stem cells ( hESCs ) remain largely undefined . Here , we demonstrate an essential role for the serine/threonine protein kinase mammalian target of rapamycin ( P42345 ) in regulating hESC long-term undifferentiated growth . Inhibition of P42345 impairs pluripotency , prevents cell proliferation , and enhances mesoderm and endoderm activities in hESCs . At the molecular level , P42345 integrates signals from extrinsic pluripotency-supporting factors and represses the transcriptional activities of a subset of developmental and growth-inhibitory genes , as revealed by genome-wide microarray analyses . Repression of the developmental genes by P42345 is necessary for the maintenance of hESC pluripotency . These results uncover a novel signaling mechanism by which P42345 controls fate decisions in hESCs . Our findings may contribute to effective strategies for tissue repair and regeneration . Dietary phytochemicals alter epigenetic events and signaling pathways for inhibition of metastasis cascade : phytoblockers of metastasis cascade . Cancer metastasis is a multistep process in which a cancer cell spreads from the site of the primary lesion , passes through the circulatory system , and establishes a secondary tumor at a new nonadjacent organ or part . Inhibition of cancer progression by dietary phytochemicals ( DPs ) offers significant promise for reducing the incidence and mortality of cancer . Consumption of DPs in the diet has been linked to a decrease in the rate of metastatic cancer in a number of preclinical animal models and human epidemiological studies . DPs have been reported to modulate the numerous biological events including epigenetic events ( noncoding micro-RNAs , histone modification , and DNA methylation ) and multiple signaling transduction pathways ( Wnt/β-catenin , Notch , Sonic hedgehog , P35354 , P00533 , MAPK- P29323 , JAK- P35610 , Akt/PI3K/ P42345 , NF-κB , AP-1 , etc. ) , which can play a key role in regulation of metastasis cascade . Extensive studies have also been performed to determine the molecular mechanisms underlying antimetastatic activity of DPs , with results indicating that these DPs have significant inhibitory activity at nearly every step of the metastatic cascade . DPs have anticancer effects by inducing apoptosis and by inhibiting cell growth , migration , invasion , and angiogenesis . Growing evidence has also shown that these natural agents potentiate the efficacy of chemotherapy and radiotherapy through the regulation of multiple signaling pathways . In this review , we discuss the variety of molecular mechanisms by which DPs regulate metastatic cascade and highlight the potentials of these DPs as promising therapeutic inhibitors of cancer . S-sulfhydration of Q02750 leads to P09874 activation and DNA damage repair . The repair of DNA damage is fundamental to normal cell development and replication . Hydrogen sulfide ( H2S ) is a novel gasotransmitter that has been reported to protect cellular aging . Here , we show that H2S attenuates DNA damage in human endothelial cells and fibroblasts by S-sulfhydrating Q02750 at cysteine 341 , which leads to P09874 activation . H2S-induced Q02750 S-sulfhydration facilitates the translocation of phosphorylated P27361 /2 into nucleus , where it activates P09874 through direct interaction . Mutation of Q02750 cysteine 341 inhibits P29323 phosphorylation and P09874 activation . In the presence of H2S , activated P09874 recruits P18887 and P49916 to DNA breaks to mediate DNA damage repair , and cells are protected from senescence . Cardiac channelopathies associated with infantile fatal ventricular arrhythmias : from the cradle to the bench . BACKGROUND : Fatal ventricular arrhythmias in the early period of life have been associated with cardiac channelopathies for decades , and postmortem analyses in P22304 victims have provided evidence of this association . However , the prevalence and functional properties of cardiac ion channel mutations in infantile fatal arrhythmia cases are not clear . METHODS AND RESULTS : Seven infants with potentially lethal arrhythmias at age < 1 year ( 5 males , age of onset 44.1 ± 72.1 days ) were genetically analyzed for P51787 , Q12809 , P15382 -5 , P63252 , Q14524 , P36382 , and P62158 by using denaturing high-performance liquid chromatography and direct sequencing . Whole-cell currents of wildtype and mutant channels were recorded and analyzed in Chinese hamster ovary cells transfected with Q14524 and Q12809 cDNA . In 5 of 7 patients , we identified 4 mutations ( p.N1774D , p.T290fsX53 , p.F1486del and p.N406K ) in Q14524 , and 1 mutation ( p.G628D ) in Q12809 . N1774D , F1486del , and N406K in Q14524 displayed tetrodotoxin-sensitive persistent late Na(+) currents . By contrast , Q14524 -T290fsX53 was nonfunctional . Q12809 -G628D exhibited loss of channel function . CONCLUSION : Genetic screening of 7 patients was used to demonstrate the high prevalence of cardiac channelopathies . Functional assays revealed both gain and loss of channel function in Q14524 mutations , as well as loss of function associated with the Q12809 mutation . DB06287 downregulates P38936 without altering cyclin D1 expression and induces autophagy and synergizes with vorinostat in mantle cell lymphoma . OBJECTIVE : To investigate the mechanisms of antiproliferative effect induced by the mammalian target of rapamycin ( P42345 ) inhibitor temsirolimus in mantle cell lymphoma ( Q8WXI8 ) . MATERIALS AND METHODS : The antiproliferative effect of temsirolimus on three well-defined Q8WXI8 cell lines was examined by the MTS assay . Induction of cell-cycle arrest , autophagy , and apoptosis were determined by fluorescence-activated cell sorting analysis . The molecular mechanisms underlining these effects were determined by Western blot . Synergy between temsirolimus and vorinostat were examined by MTS assay and the combination index was calculated . RESULTS : DB06287 has antiproliferative activity in three Q8WXI8 cell lines in a dose- and time-dependent manner . Mechanistically , temsirolimus inhibited P42345 , as evidenced by inhibition of ribosomal S6 phosphorylation , and induced cell-cycle arrest in the G(0)/G(1) phase and a decrease in P38936 expression without altering p27 or cyclin D1 levels . Furthermore , temsirolimus increased the number of acidic vesicular organelles and the amount of microtubule-associated protein 1 light-chain 3 processing , which are characteristic of autophagy , without induction of apoptosis . These changes were not associated with alteration in phosphorylated extracellular signal-regulated kinase ( P29323 ) , beclin-1 , Bax , or Bak levels . In contrast , treatment of these cell lines with the histone deacetylase inhibitor vorinostat decreased P29323 phosphorylation , activated caspase 3 , and induced apoptosis . Moreover , temsirolimus synergized with submaximal concentrations of vorinostat in all Q8WXI8 cell lines . CONCLUSION : This is the first report of temsirolimus-induced autophagy in Q8WXI8 , and of vorinostat inhibition of P29323 phosphorylation in Q8WXI8 . Collectively , these data suggest that the combination of temsirolimus and vorinostat have synergistic antiproliferative activity in Q8WXI8 cells by distinctively targeting apoptosis and autophagy . Systematic meta-analyses and field synopsis of genetic association studies in schizophrenia : the SzGene database . In an effort to pinpoint potential genetic risk factors for schizophrenia , research groups worldwide have published over 1,000 genetic association studies with largely inconsistent results . To facilitate the interpretation of these findings , we have created a regularly updated online database of all published genetic association studies for schizophrenia ( ' SzGene ' ) . For all polymorphisms having genotype data available in at least four independent case-control samples , we systematically carried out random-effects meta-analyses using allelic contrasts . Across 118 meta-analyses , a total of 24 genetic variants in 16 different genes ( P02649 , P21964 , DAO , P21728 , P14416 , P21917 , Q96EV8 , P47870 , Q13224 , HP , P01584 , P42898 , O75051 , P31645 , P04637 and P17752 ) showed nominally significant effects with average summary odds ratios of approximately 1.23 . Seven of these variants had not been previously meta-analyzed . According to recently proposed criteria for the assessment of cumulative evidence in genetic association studies , four of the significant results can be characterized as showing ' strong ' epidemiological credibility . Our project represents the first comprehensive online resource for systematically synthesized and graded evidence of genetic association studies in schizophrenia . As such , it could serve as a model for field synopses of genetic associations in other common and genetically complex disorders . Pulmonary arterial dysfunction in insulin resistant obese Zucker rats . BACKGROUND : P01308 resistance and obesity are strongly associated with systemic cardiovascular diseases . Recent reports have also suggested a link between insulin resistance with pulmonary arterial hypertension . The aim of this study was to analyze pulmonary vascular function in the insulin resistant obese Zucker rat . METHODS : Large and small pulmonary arteries from obese Zucker rat and their lean counterparts were mounted for isometric tension recording . mRNA and protein expression was measured by RT-PCR or Western blot , respectively . KV currents were recorded in isolated pulmonary artery smooth muscle cells using the patch clamp technique . RESULTS : Right ventricular wall thickness was similar in obese and lean Zucker rats . Lung Q13873 , KV1.5 and 5- Q13049 receptor mRNA and protein expression and KV current density were also similar in the two rat strains . In conductance and resistance pulmonary arteries , the similar relaxant responses to acetylcholine and nitroprusside and unchanged lung P29474 expression revealed a preserved endothelial function . However , in resistance ( but not in conductance ) pulmonary arteries from obese rats a reduced response to several vasoconstrictor agents ( hypoxia , phenylephrine and 5-HT ) was observed . The hyporesponsiveness to vasoconstrictors was reversed by L-NAME and prevented by the P35228 inhibitor 1400W . CONCLUSIONS : In contrast to rat models of type 1 diabetes or other mice models of insulin resistance , the obese Zucker rats did not show any of the characteristic features of pulmonary hypertension but rather a reduced vasoconstrictor response which could be prevented by inhibition of P35228 . Protective effects of mineralocorticoid receptor blockade against neuropathy in experimental diabetic rats . AIMS : P08235 ( MR ) blockade is an effective treatment for hypertension and diabetic nephropathy . There are no data on the effects of MR blockade on diabetic peripheral neuropathy ( DPN ) . The aim of this study was to determine whether MRs are present in the peripheral nerves and to investigate the effectiveness of MR blockade on DPN in streptozotocin ( Q11206 ) -induced diabetic rats . METHODS : Expression of MR protein and messenger RNA ( mRNA ) was examined in the peripheral nerves using Western blot analysis and RT-PCR . We next studied the effects of the selective MR antagonist eplerenone and the angiotensin II receptor blocker candesartan on motor and sensory nerve conduction velocity ( NCV ) , morphometric changes and cyclooxygenase-2 ( P35354 ) gene and NF-κB protein expression in the peripheral nerves of Q11206 -induced diabetic rats . RESULTS : Expression of MR protein and mRNA in peripheral nerves was equal to that in the kidney . Motor NCV was significantly improved by 8 weeks of treatment with either eplerenone ( Q04695 ± 1.2 m/s ) or candesartan ( 46.4 ± 6.8 m/s ) compared with control diabetic rats ( 33.7 ± 2.0 m/s ) ( p < 0.05 ) . Sensory NCV was also improved by treatment with candesartan or eplerenone in diabetic rats . DB00700 and candesartan caused significant improvement in mean myelin fibre area and mean myelin area compared with control diabetic rats ( p < 0.05 ) . P35354 mRNA and NF-κB protein were significantly elevated in the peripheral nerves of diabetic rats compared with control rats , and treatment with eplerenone or candesartan reduced these changes in gene expression ( p < 0.05 ) . CONCLUSION : MR blockade may have neuroprotective effects on DPN . Differential role for phospholipase D1 and phospholipase D2 in 12-O-tetradecanoyl-13-phorbol acetate-stimulated MAPK activation , Cox-2 and P10145 expression . Phospholipase D ( PLD ) is expressed in many tissues and stimulated by growth factors and cytokines . However , the role of PLD in signal transduction is still not well-understood . Human embryonic kidney ( P29320 -293 ) cells exhibit low levels of both Q13393 and O14939 mRNA , however , only Q13393 protein was detected by Western blot . When either isoform of PLD was stably expressed in P29320 -293 cells , we observed an increased PLD activity in a cell-free system and a 12-O-tetradecanoyl-13-phorbol acetate ( TPA ) -stimulated increase in PLD activity in intact cells . This system was then used to elucidate the effects of PLD activity on TPA-stimulated signaling pathways . Two such pathways , the mitogen-activated protein kinases ( MAPK ) , extracellular regulated protein kinase ( P29323 ) and p38 are activated by growth factors and cellular stress , respectively . We found that TPA stimulated P29323 phosphorylation regardless of the expression status of PLD . In contrast to P29323 kinase , P29320 -293 cells were unable to induce p38 phosphorylation by TPA stimulation . When P29320 -293 cells expressed either Q13393 or O14939 , we observed elevated p38 phosphorylation in response to TPA stimulation . The P29323 and p38 MAPKs can also stimulate the expression of both cyclooxygenase-2 ( Cox-2 ) and interleukin-8 ( P10145 ) . We used this system to differentiate the effect of Q13393 or O14939 activity on the expression of Cox-2 and P10145 . Increased Cox-2 and P10145 expression was found only in P29320 -293 cells expressing Q13393 . These data identify a novel role for the Q13393 isoform in the induction of gene expression and provide new insight into the differential role of Q13393 and O14939 in cells . P08235 antagonism in the treatment of chronic central serous chorioretinopathy : a pilot study . PURPOSE : Based on experimental data showing that central serous chorioretinopathy could result from overactivation of mineralocorticoid receptor pathway in choroid vessels , the authors studied eplerenone , a mineralocorticoid receptor antagonist , as a potential treatment for chronic central serous chorioretinopathy . METHODS : This nonrandomized pilot study included 13 patients with central serous chorioretinopathy of at least 4-month duration , treated with 25 mg/day of oral eplerenone for a week followed by 50 mg/day for 1 or 3 months . The primary outcome measure was the changes in central macular thickness recorded by optical coherence tomography , and the secondary outcomes included changes in foveal subretinal fluid ( P11831 ) measured by O75051 , in best-corrected visual acuity ( BCVA ) and the percentage of eyes achieving complete resolution of subretinal fluid during the treatment period . RESULTS : Central macular thickness decreased significantly from 352 ± 139 μm at baseline to 246 ± 113 μm and 189 ± 99 μm at 1 and 3 months under eplerenone treatment ( P < 0.05 and P < 0.01 , respectively ) . At 3 months , the subretinal fluid significantly decreased compared with baseline subretinal fluid ( P < 0.01 ) and best-corrected visual acuity significantly improved compared with baseline best-corrected visual acuity ( P < 0.001 ) . CONCLUSION : DB00700 treatment was associated with a significant reduction in central macular thickness , subretinal fluid level , and an improvement in visual acuity . Randomized controlled trials are needed to confirm these encouraging results . Effect of dietary NaCl on tyrosine hydroxylase in the superior cervical ganglia of Dahl rats . To investigate the involvement of peripheral catecholamines in the development of Dahl-Iwai salt-sensitive ( Q8IX12 /Eis ) hypertension , we performed immunohistochemical staining of tyrosine hydroxylase ( TH ) in the superior cervical ganglia ( SCG ) of Q8IX12 /Eis rats and Dahl-Iwai salt-resistant ( P30518 /Eis ) rats , and in situ hybridization histochemistry for demonstration of TH mRNA localization in the SCG of these rats . Q8IX12 /Eis and P30518 /Eis rats were fed on a high ( 8 % ) salt diet or on a low ( 0.3 % ) salt diet for 4 weeks . Nerve cells in the SCG of Q8IX12 /Eis high salt rats exhibited more intense TH-immunoreactivity ( P < 0.01 ) and hybridization signals ( P < 0.01 ) than those of the other experimental groups . These findings suggest that activation of peripheral sympathetic nerves may account for hypertension in Q8IX12 /Eis rats on a high salt diet . Performance of in silico analysis in predicting the effect of non-synonymous variants in inherited steroid metabolic diseases . BACKGROUND : Unclassified genetic variants are commonly encountered in molecular diagnostic service . In silico analyses using web-based predictive programs may provide information on the nature of the genetic variants , and help to prioritize novel variants for in vitro functional characterization . The objective of this study was to compare the performance of three such programs in genes related to steroid metabolism . METHODS : The effects of non-synonymous benign and pathogenic sequence variants in the P05108 , P15538 , P19099 , P05093 , P11511 , P08686 , Q9UBM7 , P26439 , P80365 , P37058 , P16435 , and P31213 genes listed in the Human Gene Mutation Database and dbSNP were tested by SIFT , PolyPhen-2 and P27169 -P . Their concordance , sensitivity , specificity , positive and negative predictive values and accuracy were assessed , using the reported phenotype and the in vitro functional data as gold standards . RESULTS : 797 sequence variants were tested . SIFT and PolyPhen-2 had high concordance , with PolyPhen-2 being slightly superior to SIFT in all assessments . P27169 -P behaved differently , with one-third of the variants unclassified . CONCLUSIONS : SIFT and PolyPhen-2 behaved similarly while P27169 -P behaved differently in predicting pathogenicity in genes related to steroid metabolism . Molecular pathologists should verify the performance of these programs before considering them in clinical decision making , and be aware that these programmes can not replace in vitro function studies . Clinicians and patients should also be informed about the limitations of genetic testing , particularly when a novel variant is encountered . P01308 suppresses IKs ( P51787 / P15382 ) currents , which require β-subunit P15382 . Abnormal QT prolongation in diabetic patients has become a clinical problem because it increases the risk of lethal ventricular arrhythmia . In an animal model of type 1 diabetes mellitus , several ion currents , including the slowly activating delayed rectifier potassium current ( IKs ) , are altered . The IKs channel is composed of P51787 and P15382 subunits , whose genetic mutations are well known to cause long QT syndrome . Although insulin is known to affect many physiological and pathophysiological events in the heart , acute effects of insulin on cardiac ion channels are poorly understood at present . This study was designed to investigate direct electrophysiological effects of insulin on IKs ( P51787 / P15382 ) currents . P51787 and P15382 were co-expressed in Xenopus oocytes , and whole cell currents were measured by a two-microelectrode voltage-clamp method . Acute application of insulin suppressed the P51787 / P15382 currents and phosphorylated Akt and extracellular signal-regulated kinase ( P29323 ) , the two major downstream effectors , in a concentration-dependent manner . Wortmannin ( 10(-6) M ) , a phosphoinositide 3-kinase ( PI3K ) inhibitor , attenuated the suppression of the currents and phosphorylation of Akt by insulin , whereas U0126 ( 10(-5) M ) , a mitogen-activated protein kinase kinase ( MEK ) inhibitor , had no effect on insulin-induced suppression of the currents . In addition , insulin had little effect on P51787 currents without P15382 , which indicated an essential role of P15382 in the acute suppressive effects of insulin . Mutagenesis studies revealed amino acid residues 111-118 within the distal third C-terminus of P15382 as an important region . P01308 has direct electrophysiological effects on IKs currents , which may affect cardiac excitability . Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 ) and cyclooxygenase ( P36551 ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg/day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq/day ) , and the experimental group was supplied with a higher sodium diet ( 2. /day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 was increased in the inner medulla . Neither the expression of P29474 nor that of P35228 was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 was increased . Neither the expression of P16066 or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 was increased in the inner medulla , while that of P23219 remained unchanged . In conclusion , the upregulation of P29475 , P01160 , and P35354 may be causally related with the aldosterone escape . Role of cardiovascular aldosterone in hypertension . DB04630 plays an important role in the pathogenesis of cardiovascular disease . We have reported that aldosterone is synthesized in cardiovascular tissues and local aldosterone synthesis plays important roles for hypertension and cardiac hypertrophy . High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP . Plasma aldosterone concentrations and P06703 were decreased by high salt intake in SHRSP . DB04630 production , the expression of P19099 mRNA and angiotensin II receptor AT1R mRNA in blood vessels were significantly increased by high salt intake . These results suggest that high salt intake increases aldosterone production and expression of the AT1R mRNA in the vascular tissue in SHRSP , which may contribute to the development of malignant hypertension in salt-loaded SHRSP . However , there are several reports of conflicting data . P08235 ( MR ) binding is tightly regulated by the enzyme 11beta-hydroxysteroid dehydrogenase type 2 ( 11beta-HSD2 ) which selectively metabolizes glucocorticoids to inactive metabolites , thus allowing for MR activation by aldosterone . We have reported that decreased 11beta-HSD2 in blood vessels in Dahl salt-sensitive ( DS ) rats , a model for salt-sensitive hypertension . Local aldosterone excess may play a significant role in the salt sensitivity and development of hypertension . High sodium intake decreased circulating rennin-angiotensin-aldosterone system and increased blood pressure and cardiac hypertrophy in DS rats , which were prevented by the treatment with a selective MR antagonist , eplerenone . DB00700 also improved endothelial nitric oxide synthase ( P29474 ) activity and P29474 mRNA expression in blood vessels in DS rats . These results further suggest that not only circulating aldosterone but also local aldosterone is of critical importance in the pathophysiology of cardiovascular disorders . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . Serial assessment of vessel interactions after drug-eluting stent implantation in unprotected distal left main coronary artery disease using frequency-domain optical coherence tomography . OBJECTIVES : This study sought to assess stent-vessel interactions after drug-eluting stent ( DES ) implantation in unprotected left main coronary artery ( ULM ) by frequency-domain optical coherence tomography ( FD- O75051 ) . BACKGROUND : Percutaneous coronary intervention using DES in ULM has been increasingly performed in routine practice . Recently , FD- O75051 assessments of DES-vessel interactions have been used as surrogates for DES safety ; however , there are no FD- O75051 studies in ULM . METHODS : We prospectively enrolled 33 consecutive patients with ULM disease treated with sirolimus- ( n = 11 ) and everolimus-eluting stents ( n = 22 ) . FD- O75051 assessments were performed post-percutaneous coronary intervention and at 9-month follow-up . Three different segments of ULM were compared : distal ( Q8IX12 ) , bifurcation ( BIF ) , and ostial-body ( BODY ) . The primary endpoints were percentages of uncovered and malapposed struts at 9-month follow-up , and the secondary endpoint was neointimal hyperplasia area . RESULTS : We analyzed 25,873 stent struts . Significant differences were demonstrated for percentage of uncovered struts ( 3.4 % , 11.7 % , and 18.7 % , respectively for Q8IX12 , BIF , and BODY ; p < 0.05 for all the comparisons ) . Malapposition was also more common in BODY ( 5.3 % ) than in Q8IX12 ( 0.6 % ) and BIF ( 2.0 % ) segments ( p < 0.05 for BODY vs. Q8IX12 , and BODY vs. BIF ) . Equivalent neointimal hyperplasia areas were demonstrated in all segments . Acute malapposition rates led to different patterns of DES-vessel interactions at 9-month follow-up . CONCLUSIONS : Distinct patterns of DES-vessel interactions were demonstrated in different segments of ULM . Acute stent strut malapposition affects these findings . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . Regulation of male fertility by P13569 and implications in male infertility . BACKGROUND : The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -activated Cl(-) and HCO(3)(-) conducting channel , mutations of which are known to be associated with male infertility . However , the underlying mechanisms remain elusive . METHODS : Literature databases were searched for papers on the topics related to P13569 and male fertility and infertility with relevant keywords . Unpublished data from authors ' laboratory were also included for analysis . RESULTS : Clinical evidence shows increased mutation frequency or reduced P13569 expression in men with congenital bilateral absence of vas deferens ( CBAVD ) or sperm abnormalities , such as azoospermia teratospermia and oligoasthenospermia . Studies on primary rodent Sertoli cells and germ cells , as well as testes from P13569 knockout mice or a cryptorchidism model , yield findings indicating the involvement of P13569 in spermatogensis through the HCO(3)(-)/ Q96PN6 / DB02527 /CREB( Q03060 ) pathway and the NF-κB/ P35354 /PGE(2) pathway . Evidence also reveals a critical role of P13569 in sperm capacitation by directly or indirectly mediating HCO(3)(-) entry that is essential for capacitation . P13569 is emerging as a versatile player with roles in mediating different signaling pathways pertinent to various reproductive processes , in addition to its long-recognized role in electrolyte and fluid transport that regulates the luminal microenvironment of the male reproductive tract . CONCLUSIONS : P13569 is a key regulator of male fertility , a defect of which may result in different forms of male infertility other than CBAVD . It would be worthwhile to further investigate the potential of developing novel diagnostic and contraceptive methods targeting P13569 . Studying sudden and unexpected infant deaths in a time of changing death certification and investigation practices : evaluating sleep-related risk factors for infant death in New York City . We describe an approach for quantifying and characterizing the extent to which sudden and unexpected infant deaths ( SUIDs ) result from unsafe sleep environments ( e.g. , prone position , bedsharing , soft bedding ) ; and present data on sleep-related infant deaths in NYC . Using a combination of vital statistics and medical examiner data , including autopsy and death scene investigation findings , we analyzed any death due to accidental threat to breathing ( ATB ) ( ICD-10 W75 & W84 ) , and deaths of undetermined intent ( Q05707 ) ( Y10-Y34 ) between 2000 and 2003 in NYC for the presence of sleep-related factors ( P11831 ) . Homicide deaths were excluded as were P22304 , since in NYC P22304 is not a certification option if environmental factors were possibly contributors to the death . All 19 ATB and 69 ( 75 % ) Q05707 had SRFs as per the OCME investigation . Black infants and infants born to teen mothers had higher P11831 death rates for both ATB and Q05707 deaths . Bedsharing was the most common P11831 ( 53 % -ATB ; 72 % - Q05707 deaths ) ; the majority of non-bedsharing infants were found in the prone position ( 60 % -ATB ; 78 % - Q05707 deaths ) . We found a high prevalence of SRFs among ATB and Q05707 deaths . This is the first local study to illustrate the importance of knowing how SUIDs are certified in order to ascertain the prevalence of infant deaths with SRFs . Advancing the research requires clarity on the criteria used by local medical examiners to categorize SUIDs . This will help jurisdictions interpret their infant mortality statistics , which in turn will improve education and prevention efforts . The V2 vasopressin receptor stimulates P27361 /2 activity independently of heterotrimeric G protein signalling . The V2 vasopressin receptor ( P30518 ) activates the mitogen activated protein kinases ( MAPK ) P27361 /2 through a mechanism involving the scaffolding protein beta arrestin . Here we report that this activating pathway is independent of G alpha s , G alpha i , G alpha q or G betagamma and that the P30518 -mediated activation of G alpha s inhibits P27361 /2 activity in a DB02527 /PKA-dependent manner . In the HEK293 cells studied , the beta arrestin-promoted activation was found to dominate over the PKA-mediated inhibition of the pathway , leading to a strong vasopressin-stimulated P27361 /2 activation . Despite the strong MAPK activation and in contrast with other GPCR , P30518 did not induce any significant increase in DNA synthesis , consistent with the notion that the stable interaction between P30518 and beta arrestin prevents signal propagation to the nucleus . Beta arrestin was found to be essential for the P27361 /2 activation , indicating that the recruitment of the scaffolding protein is necessary and sufficient to initiate the signal in the absence of any other stimulatory cues . Based on the use of selective pharmacological inhibitors , dominant negative mutants and siRNA , we conclude that the beta arrestin-dependent activation of P27361 /2 by the P30518 involves c-Src and a metalloproteinase-dependent trans-activation event . These findings demonstrate that beta arrestin is a genuine signalling initiator that can , on its own , engage a MAPK activation machinery upon stimulation of a GPCR by its natural ligand . In vitro investigations into the roles of drug transporters and metabolizing enzymes in the disposition and drug interactions of dolutegravir , a HIV integrase inhibitor . DB08930 ( DTG ; S/GSK1349572 ) is a potent HIV-1 integrase inhibitor with a distinct resistance profile and a once-daily dose regimen that does not require pharmacokinetic boosting . This work investigated the in vitro drug transport and metabolism of DTG and assessed the potential for clinical drug-drug interactions . DTG is a substrate for the efflux transporters P-glycoprotein ( Pgp ) and human breast cancer resistance protein ( Q9UNQ0 ) . Its high intrinsic membrane permeability limits the impact these transporters have on DTG 's intestinal absorption . UDP-glucuronosyltransferase ( P78381 ) 1A1 is the main enzyme responsible for the metabolism of DTG in vivo , with cytochrome P450 ( P450 ) 3A4 being a notable pathway and P35503 and O60656 being only minor pathways . DTG demonstrated little or no inhibition ( IC(50) values > 30 μM ) in vitro of the transporters Pgp , Q9UNQ0 , multidrug resistance protein 2 , organic anion transporting polypeptide 1B1/3 , organic cation transporter ( O75051 ) 1 , or the drug metabolizing enzymes P05177 , 2A6 , 2B6 , 2C8 , 2C9 , 2C19 , 2D6 , 3A4 , P22309 , or 2B7 . Further , DTG did not induce P05177 , 2B6 , or 3A4 mRNA in vitro using human hepatocytes . DTG does inhibit the renal OCT2 ( IC(50) = 1.9 μM ) transporter , which provides a mechanistic basis for the mild increases in serum creatinine observed in clinical studies . These in vitro studies demonstrate a low propensity for DTG to be a perpetrator of clinical drug interactions and provide a basis for predicting when other drugs could result in a drug interaction with DTG . Reduction of neuronal and inducible nitric oxide synthase gene expression in patients with cystic fibrosis . As a consequence of diminished nitric oxide synthase ( NOS ) protein concentration , the airway concentration of nitric oxide ( NO ) is reduced in patients with cystic fibrosis ( CF ) . This appears to lead to a reduced elimination of such microorganisms as Pseudomonas aeruginosa . The objective of this study was to analyze whether inducible ( P35228 ) , endothelial ( P29474 ) and neuronal ( P29475 ) NOS are reduced at mRNA level and if so whether this is caused directly by the defective CF transmembrane conductance regulator ( P13569 ) . Nasal polyps from three patients with CF and four otherwise healthy patients were obtained . The expression of the three NOS isoenzymes was quantified using real-time PCR . The P35228 expression was assessed in colon carcinoma cells ( CaCo ) transfected with a normal and a mutated ( DeltaF508 ) P13569 . In CF patients , P35228 mRNA expression was 10-to 20-fold and P29475 gene expression was one-fifth to one-tenth that in control patients ( P < 0.001 ) . In CaCo cells , P35228 gene expression under basal and endotoxin-stimulated conditions did not differ between cells transfected with a mutated P13569 and those transfected with an intact P13569 . This observation suggests that cystic fibrosis is associated with reduced P35228 and P29475 gene expression in nasopharyngeal tissue , possibly disturbing the barrier against infective agents already at the site of entrance . Enhanced NF-κB activity impairs vascular function through P09874 - , SP-1- , and P35354 -dependent mechanisms in type 2 diabetes . Type 2 diabetes ( T2D ) is associated with vascular dysfunction . We hypothesized that increased nuclear factor-κB ( NF-κB ) signaling contributes to vascular dysfunction in T2D . We treated type 2 diabetic ( db(-)/db(-) ) and control ( db(-)/db(+) ) mice with two NF-κB inhibitors ( 6 mg/kg dehydroxymethylepoxyquinomicin twice a week and 500 μg/kg/day IKK-NBD peptide ) for 4 weeks . Pressure-induced myogenic tone was significantly potentiated , while endothelium-dependent relaxation ( EDR ) was impaired in small coronary arterioles and mesenteric resistance artery from diabetic mice compared with controls . Interestingly , diabetic mice treated with NF-κB inhibitors had significantly reduced myogenic tone potentiation and improved EDR . Importantly , vascular function was also rescued in db(-)/db(-p50NF-κB-/-) and db(-)/db(- P09874 -/-) double knockout mice compared with db(-)/db(-) mice . Additionally , the acute in vitro downregulation of NF-κB-p65 using p65NF-κB short hairpin RNA lentivirus in arteries from db(-)/db(-) mice also improved vascular function . The NF-κB inhibition did not affect blood glucose level or body weight . The RNA levels for Sp-1 and P29474 phosphorylation were decreased , while p65NF-κB phosphorylation , cleaved poly(ADP-ribose) polymerase ( PARP ) -1 , and cyclooxygenase ( P36551 ) -2 expression were increased in arteries from diabetic mice , which were restored after NF-κB inhibition and in db(-)/db(-p50NF-κB-/-) and db(-)/db(- P09874 -/-) mice . In the current study , we provided evidence that enhanced NF-κB activity impairs vascular function by P09874 - , Sp-1- , and P35354 -dependent mechanisms in male type 2 diabetic mice . Therefore , NF-κB could be a potential target to overcome diabetes-induced vascular dysfunction . Acute heat stress induces edema and nitric oxide synthase upregulation and down-regulates mRNA levels of the Q05586 , Q12879 and Q13224 subunits in the rat hippocampus . The influence of heat stress on constitutive isoform of neuronal nitric oxide synthase ( P29474 ) and DB01221 receptor gene expression in hippocampus was examined in a rat model . Subjection of animals to 4 h heat stress at 38 degrees C resulted in a marked upregulation of P29474 in the hippocampus accompanied with a marked general expansion and edematous cell changes . On the other hand DB01221 receptor messenger RNA encoding Q05586 , Q12879 and Q13224 subunits showed a marked downregulation in the hippocampus of heat stressed rats compared to the controls . Our results show that upregulation of P29474 is instrumental in heat stress associated edema and cell injury . Furthermore , an increased production of NO as evident with upregulation of P29474 appears to be a key factor in the downregulation of DB01221 receptor gene expression in heat stress . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . Uptake of DB06704 and catecholamines in noradrenaline- and organic cation transporter-expressing cells : potential use of corticosterone for a preferred uptake in neuroblastoma- and pheochromocytoma cells . For imaging of neuroblastoma and phaeochromocytoma , [(123)I]meta-iodobenzylguanidine ( [(123)I] DB06704 ) is routinely used , whereas [(18)F]6-fluorodopamine ( [(18)F]6-FDA ) is sporadically applied for positron emission tomography in pheochromocytoma . Both substances are taken up by catecholamine transporters ( CATs ) . In competition , some other cell types are able to take up catecholamines and related compounds probably by organic cation ( O75051 ) [ extraneuronal monoamine ( EMT ) ] transporters ( OCT1 , OCT2 , OCT3=EMT ) . In this study , we investigated the uptake of radioiodine-labeled meta-iodobenzylguanidine ( DB06704 ) as well as [(3)H]dopamine ( mimicring 6-fluorodopamine ) and [(3)H]noradrenaline . SK-N-SH ( neuroblastoma ) and PC-12 ( phaeochromocytoma ) cells were used and compared with P29320 -293 cells transfected with OCT1 , OCT2 and OCT3 , respectively . In order to gain a more selective uptake in CAT expressing tumor cells , different specific inhibitors were measured . Uptake of DB06704 into O75051 -expressing cells was similar or even better as into both CAT-expressing cell lines , whereas dopamine and noradrenaline uptake was much lower in O75051 -expressing cells . In presence of corticosterone ( f.c . 10(-4) M ] , catecholamine and DB06704 uptake into SK-N-SH and PC-12 cells was only slightly reduced . In contrast , this process was significantly inhibited in OCT2 and OCT3 transfected P29320 -293 as well as in Caki-1 cells , which naturally express OCT3 . We conclude that the well-known corticosteroid corticosterone might be used in combination with [(18)F]6-FDA or [(123)I] DB06704 to improve specific imaging of neuroblastoma and pheochromocytoma and to reduce irradiation dose to nontarget organs in [(131)I] DB06704 treatment . Expression of P35354 and DB01221 receptor genes at the cochlea and midbrain in salicylate-induced tinnitus . OBJECTIVE/HYPOTHESIS : The expression of the genes for cyclooxygenase ( P36551 ) and DB01221 receptor ( NR ) has seldom been reported in tinnitus . We hypothesized that expression of P35354 and NR was altered in the cochlea and midbrain in salicylate-induced tinnitus . STUDY DESIGN : Experimental study on mice . METHODS : We evaluated the tinnitus score and mRNA expression levels of P35354 and NR subtype 2B ( Q13224 ) in the cochlea and midbrain in response to intraperitoneal injections of salicylate for 4 days . RESULTS : At day 4 of tinnitus induction , the mean weights of the whole body and midbrain did not change greatly in both control and salicylate groups . The tinnitus score was not elevated from day 1 to day 4 in the control group , but increased day by day in the salicylate group . The mRNA expression level of P35354 decreased slightly in the salicylate group in the cochlea ( 1.1 ± 0.33 vs. 1.3 ± 0.49 , P = .0752 ) and in the midbrain ( 0.9 ± 0.10 versus 1.0 ± 0.35 , P = .0489 ) . Inversely , the expression levels of the Q13224 gene increased moderately in the salicylate group in the cochlea ( 3.7 ± 0.47 versus 2.3 ± 1.13 , P < 0.0001 ) and in the midbrain ( 1.6 ± 0.64 versus 1.0 ± 0.44 , P = .0007 ) . CONCLUSIONS : Salicylate induced tinnitus and altered the expression of the P35354 and Q13224 genes in the cochlea and midbrain of mice . These findings might contribute to further understanding of pathophysiology and therapy of tinnitus . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability .

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interacts_with DB01427?

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Phosphodiesterase 4 inhibitor cilomilast inhibits fibroblast-mediated collagen gel degradation induced by tumor necrosis factor-alpha and neutrophil elastase . Tissue destruction , resulting in emphysema , can be a consequence of several pathologic processes . The current study evaluated the effects of the phosphodiesterase (PDE)4 inhibitor , cilomilast , and other PDE inhibitors on the ability of fibroblasts to degrade extracellular matrix . Using the three-dimensional collagen gel culture system , fibroblasts ( HFL-1 ) were cultured with tumor necrosis factor ( P01375 ) -alpha , known to induce matrix metalloproteinase ( MMP ) release , and/or neutrophil elastase ( NE ) , which can induce MMP activation . On Day 4 , gels containing P01375 and NE were significantly degraded ( 20.8 +/- 2.9 % of original collagen content ) . DB03849 ( 10 micro M ) inhibited this degradation ( 84.4 +/- 8.4 % ) . DB01427 , a PDE3 inhibitor , and zaprinast , a O76074 inhibitor , had no effect . Gelatin zymography and immunoblotting revealed that fibroblasts cultured with P01375 released increased amounts of latent P03956 and -9 . The addition of NE resulted in the conversion of P03956 and -9 to their active forms , indicative of collagen degradation . DB03849 inhibited the release of P03956 and -9 , as well as conversion of P03956 to its active form . Using real-time PCR analysis , cilomilast 's effect on P03956 release was not associated with the proteinase 's mRNA expression , suggesting that the inhibition of release is regulated at the post-transcriptional level . These results suggest that cilomilast may be a potentially effective therapeutic agent in diseases characterized by excessive tissue destruction , such as emphysema . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Desmopressin ( DB00035 ) induces NO production in human endothelial cells via V2 receptor- and DB02527 -mediated signaling . The hemostatic agent desmopressin ( DB00035 ) also has strong vasodilatory effects . DB00035 is a selective agonist for the vasopressin V2 receptor ( P30518 ) , which is coupled to DB02527 -dependent signaling . DB00035 -induced vasodilation may be due to endothelial NO synthase ( P29474 ) activation . This hypothesis implies DB02527 -mediated P29474 activation . It also implies wide extrarenal , endothelial P30518 expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 -raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 enzymatic activity , in a partly calcium-independent manner . DB02527 -mediated P29474 activation is associated with phosphorylation of residue Ser1177 , in a phosphatidyl inositol 3-kinase ( PI3K ) -independent manner . HUVECs do not express P30518 . However , after heterologous P30518 expression , DB00035 induces DB02527 -dependent P29474 activation via Ser1177 phosphorylation . We have previously found P30518 expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 and other DB02527 -raising agents can activate P29474 via PI3K-independent Ser1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 -induced vasodilation . Circulating apoptotic proteins are increased in long-term disease-free breast cancer survivors . Circulating apoptotic proteins are increased in patients with heart failure . We evaluated whether circulating soluble ( s ) apoptosis-related proteins and inflammation markers are increased in long-term disease free breast cancer survivors and associated with cardiotoxicity , and if subgroups could be identified based on the applied treatments . Circulating tumour necrosis factor ( P01375 ) alpha , sTNF-receptor ( sTNF-R ) 1 and 2 , sFas , sFas ligand , sTNF-related apoptosis inducing ligand ( sTRAIL ) and serum P04626 were measured with immunoassay . High-sensitivity P02741 ( HS-CRP ) , fibrinogen , plasma B-type and N-terminal atrial natriuretic peptide ( NT- P01160 and DB04899 ) were also determined . Thirty-four patients with median 6.0 years follow-up and 12 healthy age-matched women were enrolled . Chemotherapy , consisting of five cycles fluorouracil , epirubicin ( 90 mg/m(2) ) , cyclophosphamide ( FEC ) ( n=14 ) or four cycles FEC followed by myeloablation with high-dose carboplatin , cyclophosphamide , thiotepa ( n=20 ) , preceded irradiation and tamoxifen . Circulating apoptosis markers were higher in patients than in controls . No associations with cardiac dysfunction were observed . sFas ligand and sTRAIL were higher in the high-dose than in the standard-dose group . In conclusion , we observed increased circulating apoptotic protein levels in long-term disease-free breast cancer survivors , treated with adjuvant chemoradiotherapy , particularly after myeloablative chemotherapy . The potential relation with late cardiotoxicity of antineoplastic therapy deserves further study . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . The use of cytokine inhibitors . A new therapeutic insight into heart failure . Cytokines are being increasingly recognized as important factors in the pathogenesis and pathophysiology of heart failure . Elevated levels of circulating cytokines have been reported in patients with heart failure , and various cytokines have been shown to depress myocardial contractility in vitro and in vivo . We have recently compared the effects on cytokine production of drugs for therapy of heart failure that have different effects on survival . DB01427 , pimobendan and vesnarinone , phosphodiesterase III inhibitors that have been shown to have short term haemodynamic benefits , inhibited P01375 production . Differential modulation of the production of IL-1beta and P05231 was observed ; amrinone and pimobendan enhanced the production of IL-1beta , whereas vesnarinone did not . As inotropic agents differentially modulate cytokine production , these agents may interfere with induction of inducible nitric oxide ( NO ) synthase through an inhibition of cytokine formation . Although differential modulation of the production of NO by inotropic agents may explain their different effect in patients with heart failure , further study is necessary to reach this conclusion . We have shown that amlodipine increases the survival of mice with viral myocarditis and inhibits expression of inducible NO synthase and production of NO in vivo and in vitro . The therapeutic effect of amlodipine may in part result from inhibition of overproduction of NO . As we learn more about the pathophysiological and pathogenetic role of cytokines in heart failure , it should be possible to design better and more targeted pharmacological agents . Furthermore , the investigation of inotropic agents that are effective against the production of cytokines may help in the classification of these agents . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . Enhanced goblet cell hyperplasia in HDC knockout mice with allergic airway inflammation . BACKGROUND : DB11320 is known to have immunoregulatory roles in allergic reactions through histamine receptor 1 ( P35367 ) , P25021 , Q9Y5N1 and Q9H3N8 . However , its role in goblet cell hyperplasia in the airways of asthma patients is yet to be clarified . OBJECTIVE : This study was designed to examine the role of histamine in goblet cell hyperplasia using histamine-deficient mice ( Hdc-/- mice ) with allergic airway inflammation . METHODS : Wild-type and Hdc-/- C57BL/6 mice were sensitized with ovalbumin ( OVA ) . After a 2-week exposure to OVA , goblet cell hyperplasia was evaluated . Cell differentials and cytokines in BALF were analyzed . The mRNA levels of P98088 and Gob-5 gene were determined quantitatively . RESULTS : The number of eosinophils in BALF increased in both the sensitized wild-type mice and Hdc-/- mice with OVA inhalation . In addition , the numbers of alveolar macrophages and lymphocytes in BALF increased significantly in the sensitized Hdc-/- mice with OVA inhalation compared to the wild-type mice under the same conditions . The concentrations of P05112 ( P05112 ) , P05113 , P35225 , Interferon-gamma ( P01579 ) , tumor necrosis factor-alpha ( P01375 ) and P60568 in the BALF all increased significantly in both groups compared to those exposed to saline . In particular , the concentration of P01375 in the Hdc-/- mice exposed to OVA was significantly higher than that in the wild-type mice under the same conditions . The mRNA levels of Gob-5 and P98088 , and the ratio of the goblet cells in the airway epithelium significantly increased in Hdc-/- mice exposed to OVA compared to wild-type mice . CONCLUSIONS : These results suggested that histamine may play a regulatory role in goblet cell hyperplasia in allergic airway inflammation . DB01427 and theophylline differentially regulate cytokine and nitric oxide production in endotoxemic mice . Intracellular cyclic nucleotide levels play an important role in the regulation of several immunological processes . Since elevation of intracellular cyclic adenosine monophosphate and/or cyclic guanosine monophosphate concentration by inhibition of phosphodiesterase ( PDE ) is known to modulate the inflammatory response , we compared the effect of amrinone , an inhibitor of the PDE III isoenzyme , and of theophylline , a nonspecific PDE inhibitor , on the plasma tumor necrosis factor-alpha ( P01375 ) , interleukin-6 ( P05231 ) , interleukin-10 ( P22301 ) , and nitric oxide response in mice to intraperitoneal injection of bacterial lipopolysaccharide ( LPS ) . Intraperitoneal treatment of animals with amrinone ( 100 mg/kg ) 30 min before LPS administration decreased both plasma P05231 and P22301 concentrations in the first phase of the response , but enhanced plasma levels of these cytokines in the second part . In contrast , pretreatment of the animals with theophylline ( 100 mg/kg ) enhanced LPS-induced plasma P05231 and P22301 levels during the whole response . However , pretreatment with both PDE inhibitors resulted in a marked inhibition of LPS-evoked plasma concentrations of P01375 and nitrite/nitrate ( breakdown products of nitric oxide ) throughout the response . This study demonstrates for the first time that amrinone and theophylline possess differential , but primarily anti-inflammatory , properties during LPS-induced systemic inflammation in the mouse . Pituitary-adrenal axis regulation in P06850 -deficient mice . P06850 ( P06850 ) -deficient ( knockout ( KO ) ) mice demonstrate severely impaired adrenal responses to restraint , ether , and fasting , and lack the normal diurnal glucocorticoid ( GC ) rhythm . Here , we summarize recent studies determining the role of P06850 in augmenting plasma adrenocorticotrophic hormone ( DB01285 ) concentration after glucocorticoid withdrawal and pituitary-adrenal axis stimulation in the context of inflammation . Even though GC insufficient , basal pituitary proopiomelanocortin ( P01189 ) mRNA , DB01285 peptide content within the pituitary , and plasma DB01285 concentrations are not elevated in P06850 KO mice . P01189 mRNA content in P06850 KO mice increases following adrenalectomy , and this increase is reversed by GC , but not aldosterone , replacement . In marked contrast to the increase in P01189 mRNA , plasma DB01285 does not increase in the P06850 KO mice following adrenalectomy . Administration of P06850 to adrenalectomized P06850 KO mice results in acute , robust DB01285 secretion . Thus , loss of GC feedback can increase P01189 gene expression in the pituitary , but P06850 action is essential for increased secretion of DB01285 into the circulation . While GC secretion is impaired in P06850 KO mice after most stimuli , we have found near-normal GC responses to inflammation and systemic immune challenge . Studies in mice with P06850 and P05231 deficiency reveal that P05231 is essential for activation of the pituitary-adrenal axis during inflammatory and other stressors in the absence of P06850 . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Myocardial tumor necrosis factor-alpha secretion in hypertensive and heart failure-prone rats . Acute increases in blood pressure ( BP ) increase myocardial tumor necrosis factor ( P01375 ) -alpha production , but it is not known whether chronic hypertensive stress elevates myocardial P01375 production , possibly contributing to cardiac remodeling , decreased cardiac function , and faster progression to heart failure . BP , cardiac function , and size were evaluated in normotensive [ Sprague-Dawley ( SD ) ] , spontaneously hypertensive ( SHR ) , and spontaneously hypertensive heart failure-prone ( SHHF ) rats at 6 , 12 , 15 , and 18 mo of age and in failing SHHF . Left ventricular tissues were evaluated for secretion of bioactive P01375 and inhibition of P01375 secretion by phosphodiesterase inhibitors . All ventricles secreted bioactive and immunoreactive P01375 , but secretion decreased with age . SHR and SHHF rats secreted more P01375 than SD rats at 6 mo of age , but only failing SHHF rats secreted significantly more P01375 at 18 mo . DB01427 inhibited P01375 secretion in all rats and was less potent but more efficacious than RO-201724 in all strains . P01375 secretion correlated with BP and left ventricular mass in 6-mo-old rats , but this relationship disappeared with age . Results suggest that hypertension and/or cardiac remodeling is associated with elevated myocardial P01375 , and , although hypertension , per se , did not maintain elevated cardiac P01375 levels , SHHF rats increase P01375 production during the end stages of failure . Effect of the phosphodiesterase III inhibitor amrinone on cytokine and nitric oxide production in immunostimulated J774.1 macrophages . The level of intracellular cyclic nucleotides is a regulatory factor in a variety of immune processes . Increases in intracellular cyclic AMP ( DB02527 ) and/or cyclic GMP ( cGMP ) concentration by the inhibition of phosphodiesterase have been shown to modulate the inflammatory response . DB01427 is a clinically used positive inotropic agent which elevates intracellular DB02527 and cGMP levels by selective inhibition of the phosphodiesterase III isoenzyme . In the current study , we investigated the effect of various concentrations ( 1-300 microM ) of amrinone on lipopolysaccharide-induced production of pro- and anti-inflammatory cytokines and of nitric oxide ( NO ) in vitro . In cultured murine J774.1 macrophages , 1 ng/ml-10 microg/ml of lipopolysaccharide from Escherichia coli O55: P46977 induced production of tumor necrosis factor-alpha ( P01375 ) , interleukin-10 , and nitrite ( breakdown product of NO ) . Pretreatment of cells with amrinone caused a dose-dependent suppression of P01375 production in the concentration range of 1-100 microM . Furthermore , this drug suppressed NO production in the range of 30-300 microM . Similarly to the results in the J774.1 cells , amrinone also inhibited P01375 and NO production in the range of 10-100 microM in primary rat peritoneal macrophages . At 300 microM , but not at lower concentrations , amrinone inhibited interleukin-10 production in lipopolysaccharide-treated J774.1 macrophages . Pretreatment of the macrophages with 100 and 300 microM amrinone increased the lipopolysaccharide-elicited translocation of nuclear factor-kappa B . Taken together , our results indicate that the phosphodiesterase III inhibitor amrinone modulates the activation/production of many pro- and anti-inflammatory factors in endotoxin-stimulated cells . It remains to be further investigated how such immunomodulatory effects contribute to the clinical profile of the agent . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . A functional comparison between the P04626 (high)/ P21860 and the P04626 (low)/ P21860 dimers on heregulin-β1-induced P03956 and P14780 expression in breast cancer cells . Overexpression of P04626 correlates with more aggressive tumors and increased resistance to cancer chemotherapy . However , a functional comparison between the P04626 (high)/ P21860 and the P04626 (low)/ P21860 dimers on tumor metastasis has not been conducted . Herein we examined the regulation mechanism of heregulin- β1 ( P04196 ) -induced P03956 and -9 expression in breast cancer cell lines . Our results showed that the basal levels of P03956 and -9 mRNA and protein expression were increased by P04196 treatment . In addition , P04196 -induced P03956 and -9 expression was significantly decreased by Q02750 /2 inhibitor , U0126 but not by phosphatidylinositol 3-kinase ( PI-3K ) inhibitor , LY294002 . To confirm the role of MEK/ P29323 pathway on P04196 -induced P03956 and -9 expression , MCF7 cells were transfected with constitutively active adenoviral- MEK ( CA-MEK ) . The level of P03956 and -9 expressions was increased by CA-MEK . P03956 and -9 mRNA and protein expressions in response to P04196 were higher in P04626 overexpressed cells than in vector alone . The phosphorylation of P04626 , P21860 , P29323 , Akt , and JNK were also significantly increased in P04626 overexpressed MCF7 cells compared with vector alone . P04196 -induced P03956 and -9 expressions were significantly decreased by lapatinib , which inhibits P00533 and P04626 activity , in both vector alone and P04626 overexpressed MCF7 cells . Finally , P04196 -induced P03956 and P14780 expression was decreased by P21860 siRNA overexpression . Taken together , we suggested that P04196 -induced P03956 and P14780 expression is mediated through P21860 dependent pathway and highly expressed P04626 may be associated with more aggressive metastasis than the low expressed P04626 in breast cancer cells . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . Pharmacological modulation of myocardial tumor necrosis factor alpha production by phosphodiesterase inhibitors . Phosphodiesterase ( PDE ) inhibitors are used as therapeutic agents for management of congestive heart failure . PDE inhibitors are potent inotropic and vasodilator drugs , which have also been shown to inhibit tumor necrosis factor alpha ( P01375 ) production . P01375 is a pleiotropic cytokine that has the ability to produce cardiac depressant and other cardiovascular effects in many disease conditions . P01375 levels are elevated in patients with chronic congestive heart failure , and it is possible that P01375 may play a role in this condition . The effects of PDE inhibitors on P01375 secretion from rat heart were evaluated in this study . Rat left ventricle was minced and incubated for 4 hr with various PDE inhibitors , and the amount of P01375 secretion was evaluated by cytotoxicity assay . Ro-20 , 1724 , etazolate , amrinone , milrinone and pentoxifylline inhibited unstimulated P01375 production , with IC50 values of 1.87 , 2.07 , 13.9 , 153 and 201 microM , respectively . Lipopolysaccharide-induced P01375 secretion from rat left ventricle was also evaluated in this study . DB01427 , milrinone and pentoxifylline inhibited lipopolysaccharide-induced P01375 secretion , with IC50 values of 14.8 , 81.6 and 748 microM , respectively , whereas Ro-2D , 1724 and etazolate had no effect on lipopolysaccharide-induced P01375 secretion . These results demonstrated that P01375 was secreted from rat left ventricle after 4 hr and different pharmacological manipulations were able to inhibit the secretion of P01375 from left ventricle . These initial pharmacological results may provide an important tool for further investigation into the beneficial effects of PDE inhibitors in congestive heart failure or other conditions where P01375 levels are elevated . Bacterial translocation in cirrhotic rats stimulates P29474 -derived NO production and impairs mesenteric vascular contractility . DB00435 ( NO ) has been implicated in the arterial vasodilation and associated vascular hyporesponsiveness to vasoconstrictors observed in liver cirrhosis . Bacteria , potent activators of NO and P01375 synthesis , are found in the mesenteric lymph nodes ( MLNs ) of ascitic cirrhotic rats . Here , we investigated the impact of bacterial translocation ( BT ) to MLNs on P01375 production , vascular NO release , and contractility in the mesenteric vasculature of ascitic cirrhotic rats . Vascular response to the alpha-adrenoagonist methoxamine , which is diminished in the superior mesenteric arterial beds of cirrhotic rats , is further blunted in the presence of BT . BT promoted vascular NO release in cirrhotic rats , an effect that depended on pressure-induced shear stress and was blocked by the NO inhibitor N(omega)-nitro-L-arginine . Removing the endothelium had the same effect . Endothelial NO synthase ( P29474 ) , but not the inducible isoform ( P35228 ) , was present in mesenteric vasculature of cirrhotic rats with and without BT , and its expression was enhanced compared with controls . P01375 was induced in MLNs by BT and accumulated in parallel in the serum . This P01375 production was associated with elevated levels of tetrahydrobiopterin ( BH(4) ) , a P01375 -stimulated cofactor and enhancer of P29474 -derived NO biosynthesis and NOS activity in mesenteric vasculature . These findings establish a link between BT to MLNs and increased P01375 production and elevated BH(4) levels enhancing P29474 -derived NO overproduction , further impairing contractility in the cirrhotic mesenteric vasculature . DB01427 suppresses the synthesis of tumor necrosis factor-alpha in human mononuclear cells . P01375 -alpha ( P01375 ) exerts a wide spectrum of biological activities and contributes to the pathophysiology of septic shock . Elevated circulating levels of P01375 have also been reported in patients with severe chronic heart failure . We studied the effect of amrinone , a class III cyclic nucleotide phosphodiesterase inhibitor used in the treatment of acute heart failure , on the synthesis of P01375 in vitro . Peripheral blood mononuclear cells from healthy volunteers or cells of a permanent monoblast cell line were stimulated for 20 h with bacterial lipopolysaccharide and different doses of amrinone . P01375 production is suppressed in a dose-dependent manner to a minimum of 9 % of controls with 1000 microM of amrinone , reaching half-maximal inhibition at 80 microM amrinone . This effect appears to be mediated via DB02527 , which accumulated nearly twofold in the presence of amrinone . Suppression of P01375 synthesis by therapeutically administered phosphodiesterase inhibitors such as amrinone may contribute to their beneficial effect in the treatment of heart failure . Interleukins 1α and 1β as regulators of steroidogenesis in human NCI-H295R adrenocortical cells . Inflammatory cytokines interleukin-1 ( IL-1 ) and tumor necrosis factor-α ( P01375 -α ) regulate the activity of the hypothalamo-pituitary-adrenal ( Q9Y251 ) axis at several levels . Although hypothalamic P06850 secretion may be the primary mechanism by which these cytokines activate the Q9Y251 axis , IL-1 expression is increased within the adrenal glands in models for systemic inflammation , and IL-1 may augment adrenal glucocorticoid production . Our aim was to investigate the direct effects of IL-1α and IL-1β on adrenal steroidogenesis and expression of three key steroidogenic genes in human adrenocortical cells using the NCI-H295R cell line as a model . mRNAs encoding receptors for IL-1 , P01375 -α , and leukemia inhibitory factor ( P15018 ) were detectable in the cell line ( Affymetrix microarray analysis ) . Both IL-1α and IL-1β increased cortisol , androstenedione , dehydroepiandrosterone and DB05804 production , and the accumulation of mRNAs for steroidogenic acute regulatory protein ( STAR ) , 17α-hydroxylase/17,20-lyase ( P05093 ) and 3β-hydroxysteroid dehydrogenase 2 ( P26439 ) in these cells ( P < 0.05 for all ) . Both ILs augmented P01375 -α- and P15018 -induced STAR and P05093 mRNA accumulation , and P01375 -α-induced cortisol production ( P < 0.05 for all ) . Both ILs also increased the apoptotic index of the cells ( P < 0.05 ) , which was efficiently neutralized by their specific antibodies . The IL-induced changes in the STAR , P26439 , and P05093 protein levels were not as evident as those in the respective mRNA levels . In conclusion , the combined effect of inflammatory cytokines at the adrenal level in acute or chronic inflammatory states could significantly stimulate glucocorticoid production , and thus explain the observed discrepancy between the cortisol and DB01285 concentrations sometimes seen in sepsis and chronic inflammatory states . Role of DB05875 signaling in enhanced nociceptive sensitization and local cytokine production after incision . Substance P ( SP ) signaling facilitates nociceptive sensitization in various inflammatory and chronic pain models and we postulated that SP signaling might also contribute to the development of post-incisional hyperalgesia . These studies used mice with a deletion of the pre-protachykinin A gene ( ppt-A(-/-) ) which codes for SP to determine the role of SP signaling in post-incisional pain and in the increased cytokine and nerve growth factor ( P01138 ) expression observed in the incised skin . SP deficient ppt-A(-/-) mice displayed reduced mechanical allodynia and heat hyperalgesia compared to the wild-type ( wt ) mice at all post-incision time points , despite similar baseline values ( p < 0.001 ) . Furthermore , the P25103 antagonist LY303870 attenuated mechanical allodynia produced by incision in the wt mice ( p < 0.001 ) . Incision also up-regulated P05231 , P01375 and KC levels but not IL-1beta after 2h in the wt mice skin . However , ppt-A(-/-) mice had more skin P01138 levels 2h post-incision . Subcutaneous hind paw SP injection produced acute and transient elevations of IL-1beta , P05231 , and KC but modest elevations in P01375 levels in the wt mice . Systemic LY303870 reversed the SP-induced elevations of these cytokines . Hind paw injection of P05231 and P01138 dose dependently produced less mechanical allodynia in the ppt-A(-/-) compared to wt mice . Additionally , SP produced mechanical allodynia in a dose-dependent fashion in wt mice . Therefore , SP supports nociceptive sensitization after hind paw incision and potentially participates directly in modulating the intensity of inflammatory response in peri-incisional tissue . Experimental autoimmune encephalomyelitis in the Wistar rat : dependence of MBP-specific T cell responsiveness on P33681 costimulation . Experimental autoimmune encephalomyelitis ( EAE ) is an animal model of human multiple sclerosis that requires the activation of autoreactive T cells for the expression of pathology . EAE has been most frequently studied in the Lewis rat model as well as in several murine models of EAE including the PLJ and B10PL strains . In the present study we describe a novel model of EAE induced in the Wistar rat strain by immunization with guinea pig spinal cord antigens and pertussis toxin ( PT ) . T cell responses were induced to myelin basic protein . Autoreactive T cells could be totally blocked by the in vitro treatment with DB01281 , a protein that blocks the costimulation of autoreactive T cells . The addition of P60568 could reverse the inhibition seen in vitro with DB01281 . The effects of inhibition of P33681 costimulation were also examined by an analysis of cytokine responses and P60568 receptor on T cells . DB01281 treatment in vitro reduced the expression of P60568 receptor on T cells , enhanced T cell apoptosis and decreased the synthesis of P60568 , P01579 and P01375 . DB01281 treatment had no effect on P22301 synthesis by T cells , a cytokine implicated in the functions of regulatory T cell subsets . Overall , our studies support the rationale of P33681 blocking therapies as a potential treatment for models of multiple sclerosis . The induction of EAE in the Wistar rat provides yet another novel model in which to examine the regulation of T cell autoimmunity . Hypoxic/normoxic preconditioning increases endothelial differentiation potential of human bone marrow CD133+ cells . CD133+ cells are hemangioblasts that have capacity to generate into both hematopoietic and endothelial cells ( ECs ) . Hypoxia/normoxia has shown to be the regulator of the balance between stemness and differentiation . In this study we performed Agilent 's whole human genome oligo microarray analysis and examined the differentiation potential of the bone-marrow-derived CD133+ cells after hypoxic/normoxic preconditioning of CD133+ cells . Results showed that there was no significant increase in erythroid colony forming unit ( CFU-E ) and CFU-granulocyte , erythrocyte , monocyte , and megakaryocyte formation with cells treated under hypoxia/normoxia . However , a significant increment of EC forming unit at 24 h ( 143.2 +/- 8.0 % ) compared to 0 h ( 100 +/- 11.4 % ) was observed in CFU-EC analysis . Reverse transcription-polymerase chain reaction and immunostaining analysis showed that the differentiated cells diminished hematopoietic stem cell surface markers and acquired the gene markers and functional phenotype of ECs . The transcriptome profile revealed a cluster of 232 downregulated and 498 upregulated genes in cells treated for 24 h under hypoxia . The upregulated genes include angiogenic genes , angiogenic growth factor genes , angiogenic cytokine and chemokine genes , as well as angiogenic-positive regulatory genes , including Q14512 , PDGFB , Q16663 , P48061 , P80162 , P05231 , P21246 , O14944 , P04626 , O95136 , P11487 , Q92913 , Q99988 , P05412 , L1CAM , Q02297 , P08138 , and PDGFB . On the other hand , angiogenesis inhibitors and related genes , including P29459 , P98177 , Q9NY15 , and P16035 , are downregulated . Taken together , hypoxic/normoxic preconditioning may lead to the differentiation of CD133+ cells toward endothelial lineage , which may improve the current clinical trial studies . Effect of matrine on the expression of DB05875 receptor and inflammatory cytokines production in human skin keratinocytes and fibroblasts . Matrine is a kind of alkaloid found in certain Sophora plants , which has been extensively used in China for the treatment of viral hepatitis , cancer , cardiac diseases and skin diseases ( such as atopic dermatitis and eczema ) . It also has been confirmed that DB05875 ( SP ) and its receptor ( neurokinin-1 receptor , P25103 ) are involved in the pathogenesis of inflammatory skin disorders . So the present study was designed to investigate the effect of matrine on the expression of P25103 and cytokines production induced by SP in HaCaT cells ( a human epidermal keratinocyte cell line ) and dermal fibroblasts . In addition , cell viability was also evaluated . The results showed that matrine inhibited the expression of P25103 in HaCaT cells and fibroblasts . SP induced the production of interleukin ( IL ) -1beta , P10145 , interferon ( IFN ) -gamma , and monocyte chemotactic protein ( MCP ) -1 in both cell types . Matrine 5-100 microg/mL had little effect on cell viability . It inhibited SP-induced IL-1beta , P10145 and P13500 production in HaCaT cells and fibroblasts , while it increased the production of P01579 in HaCaT cells . Both SP and matrine had no effect on the secretion of P05231 . These findings suggest that matrine may have potential treatment function on SP related cutaneous inflammation by inhibition of the expression of DB05875 receptor and regulation of the production of inflammatory cytokines . The influence of costimulation and regulatory P01730 + T cells on intestinal IgA immune responses . It is thought that IgA B-cell differentiation is highly dependent on activated P01730 + T cells . In particular , cell-cell interactions in the Peyer 's patches involving P25942 and/or P33681 / P42081 have been implicated in germinal-center formation and IgA B-cell development . Also soluble factors , such as P05112 , P05113 , P05231 , and TGF beta may be critical for IgA B-cell differentiation in vivo . Here we report on some paradoxical findings with regard to IgA B-cell differentiation and specific mucosal immune responses that we have recently made using gene knockout mice . More specifically , we have investigated to what extent absence of P01730 + T cells , relevant cytokines , or T-cell-B-cell interactions would influence IgA B-cell differentiation in vivo . Using P01730 - or P05112 -gene knockout mice or mice made transgenic for DB01281 , we found that , although specific responses were impaired , total IgA production and IgA B-cell differentiation appeared to proceed normally . However , a poor correlation was found between , on the one hand , GC formation and IgA differentiation and , on the other hand , the ability to respond to T-cell-dependent soluble protein antigens in these mice . Thus , despite the various deficiencies in P01730 + T-cell functions seemingly intact IgA B-cell development was observed . DB00067 decreases sepsis-induced pulmonary inflammation through the P30518 . The early use of vasopressors in sepsis has been associated with a decrease in immune activation independent of hemodynamic effects , although the mechanism behind this remains unclear . We hypothesize that low dose vasopressin will reduce the pulmonary inflammation associated with sepsis . Our aims were to ( 1 ) determine whether vasopressin reduces lipopolysaccharide ( LPS ) -induced pulmonary inflammation and ( 2 ) determine which vasopressin receptor is responsible for pulmonary immune modulation . Mice were treated with intraperitoneal LPS to induce both systemic and pulmonary inflammation . DB00067 or saline was infused via peritoneal pump and interleukin 6 ( P05231 ) in lung and serum was measured at 6h . NF-kappaB activation as was determined in the lung through immunoblotting total and phospho-IkappaB . Hemodynamic data was also obtained at the 6h mark . In a separate series of experiments mice received both LPS and vasopressin infusion following pretreatment with vasopressin receptor antagonists to V1R , P30518 and OTR . Low dose LPS dramatically raises both serum P05231 and pulmonary levels of P05231 and phospho-IkappaB despite no significant changes in mean arterial pressure at 6h . Compared to saline , vasopressin infusion significantly decreases both the pulmonary P05231 levels and phospho-IkappaB in LPS treated mice without raising arterial pressure . Pretreatment with P30518 antagonist results in complete attenuation of vasopressin 's immunosuppressive effects , with restoration of pulmonary P05231 and phospho-IkappaB levels to those seen with LPS alone . CONCLUSIONS : DB00067 exerts a local anti-inflammatory effect on the lung through the P30518 in a model of sepsis .

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interacts_with DB09078?

multiple_choice

DB06643 -- an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 ) , a cytokine member of the P01375 family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . The role of HIV-related chemokine receptors and chemokines in human erythropoiesis in vitro . In order to better define the role of HIV-related chemokines in human erythropoiesis we studied : A ) the expression of chemokine receptors , both on human P28906 (+) cells which include erythroid progenitors and on more mature erythroid cells ; B ) the functionality of these receptors by calcium flux , chemotaxis assay and phosphorylation of mitogen-activated protein kinases ( MAPK ) Q8NFH3 /44 ( P27361 / P28482 ) and AKT , and finally C ) the influence of chemokines on BFU-E formation . We found that HIV-related chemokine receptor P61073 , but not P51681 , is detectable on human P28906 (+) BFU-E cells . P61073 surface expression decreased during erythroid maturation , although P61073 mRNA was still present in cells isolated from differentiated erythroid colonies . P48061 , a P61073 ligand , induced calcium flux and phosphorylation of MAPK ( Q8NFH3 /44 ) and AKT in P28906 (+) P10721 (+) bone marrow mononuclear cells which contain BFU-E , as well as chemotactic activity of both human P28906 (+) BFU-E progenitors and erythroid cells isolated from day 2-6 BFU-E colonies . Responsiveness to P48061 decreased when the cells differentiated to the point of surface expression of the erythroid-specific marker P02724 . In contrast , the P51681 ligands ( macrophage inflammatory protein-1alpha [ MIP-1alpha ] , MIP-1beta , and RANTES ) did not activate calcium flux , MAPK and AKT phosphorylation or chemotaxis of P28906 (+) P10721 (+) cells or cells isolated from the BFU-E colonies . Interestingly , none of the chemokines tested in this study had any effect on BFU-E colony formation . In conclusion , only P61073 is functional , and its specific ligand P48061 may therefore play an important role in the homing and/or retention of early erythroid precursors in the bone marrow environment . Antitumor activity of lenvatinib ( e7080 ) : an angiogenesis inhibitor that targets multiple receptor tyrosine kinases in preclinical human thyroid cancer models . Inhibition of tumor angiogenesis by blockading the vascular endothelial growth factor ( P15692 ) signaling pathway is a promising therapeutic strategy for thyroid cancer . DB09078 mesilate ( lenvatinib ) is a potent inhibitor of P15692 receptors ( P17948 -3 ) and other prooncogenic and prooncogenic receptor tyrosine kinases , including fibroblast growth factor receptors ( P11362 -4 ) , platelet derived growth factor receptor α ( PDGFRα ) , P10721 , and P07949 . We examined the antitumor activity of lenvatinib against human thyroid cancer xenograft models in nude mice . Orally administered lenvatinib showed significant antitumor activity in 5 differentiated thyroid cancer ( DTC ) , 5 anaplastic thyroid cancer ( ATC ) , and 1 medullary thyroid cancer ( P04629 ) xenograft models . DB09078 also showed antiangiogenesis activity against 5 DTC and 5 ATC xenografts , while lenvatinib showed in vitro antiproliferative activity against only 2 of 11 thyroid cancer cell lines : that is , RO82-W-1 and TT cells . Western blot analysis showed that cultured RO82-W-1 cells overexpressed P11362 and that lenvatinib inhibited the phosphorylation of P11362 and its downstream effector Q8WU20 . DB09078 also inhibited the phosphorylation of P07949 with the activated mutation C634W in TT cells . These data demonstrate that lenvatinib provides antitumor activity mainly via angiogenesis inhibition but also inhibits FGFR and P07949 signaling pathway in preclinical human thyroid cancer models . DB00104 -targeted liposomes loaded with CPT-11 enhanced cytotoxicity for the treatment of medullary thyroid carcinoma . Medullary thyroid carcinoma ( P04629 ) is a rare endocrine tumor that frequently metastasizes , but treatment with irinotecan ( CPT-11 ) is limited because of side effects . P04629 is known to overexpress the somatostatin receptor subtype 2 ( P30874 ) . DB00104 ( Oct ) is a somatostatin analogue that has a high binding affinity for SSTR and can be used as a tumor-targeting ligand . We prepared Oct-targeted liposomes loaded with CPT-11 using Oct-poly ( ethylene glycol ) ( PEG ) -lipid and evaluated Oct-mediated association and cytotoxicity of the liposomes with an P04629 cell line TT . The association of higher concentrations of modified Oct-targeted liposomes with TT cells was significantly higher than PEGylated liposomes and was significantly inhibited by empty Oct-targeted liposomes but not by free Oct . With exposure for 96 h , the cytotoxicity of Oct-targeted liposomal CPT-11 ( IC50 : 1.05 ± 0.47 μM ) was higher than free CPT-11 ( IC50 : 3.76 ± 0.61 μM ) or PEGylated liposomal CPT-11 ( IC50 : 3.05 ± 0.28 μM ) . In addition , empty Oct-targeted liposomes showed significantly higher cytotoxicity than empty nontargeted liposomes at a concentration where free Oct did not show cytotoxicity , suggesting that Oct as a ligand showed cytotoxicity . Moreover , Oct-targeted liposomal CPT-11 led to significantly higher antitumor activity and prolonged the survival time compared with nontargeted liposomal and free CPT-11 at a one-third dose and lower administration times with free CPT-11 . These findings indicated that Oct-targeted liposomes loaded with CPT-11 may offer considerable potential for P04629 chemotherapy because cytotoxicity of both CPT-11 and Oct was enhanced by effective cellular uptake via P30874 . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . [ 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 . [ Bone metastasis in prostate cancer ] . Bone metastasis and skeletal complications have a devastating impact on the quality of life and are a major cause of morbidity in prostate cancer patients . In addition to established bone-targeted therapies , new drugs such as endothelin A receptor antagonists , MET and P35968 antagonists or radiopharmaceuticals are in the focus of development . The standard care in prostate cancer patients with bone metastases to prevent skeletal-related events ( SRE ) are bisphosphonates . DB06643 , a human monoclonal antibody against O14788 , appeared to be superior to zoledronic acid for prevention of SRE and has been shown to prolong bone metastases-free survival . In contrast to zoledronic acid , denosumab clearance is not dependent on kidney function and can be administered subcutaneously . Similar rates of toxicity were observed for both substances ; however , long-term data for denosumab are limited . Distinct binding mode of multikinase inhibitor lenvatinib revealed by biochemical characterization . DB09078 is an oral multikinase inhibitor that selectively inhibits vascular endothelial growth factor ( P15692 ) receptors 1 to 3 and other proangiogenic and oncogenic pathway-related receptor tyrosine kinases . To elucidate the origin of the potency of lenvatinib in P15692 receptor 2 ( P35968 ) inhibition , we conducted a kinetic interaction analysis of lenvatinib with P35968 and X-ray analysis of the crystal structure of P35968 -lenvatinib complexes . Kinetic analysis revealed that lenvatinib had a rapid association rate constant and a relatively slow dissociation rate constant in complex with P35968 . Co-crystal structure analysis demonstrated that lenvatinib binds at its DB00171 mimetic quinoline moiety to the DB00171 binding site and to the neighboring region via a cyclopropane ring , adopting an DB00128 - DB00120 - DB00145 ( DFG ) - " in " conformation . These results suggest that lenvatinib is very distinct in its binding mode of interaction compared to the several approved P35968 kinase inhibitors . Regulation of gene expression in melanoma : new approaches for treatment . The molecular changes associated with the transition of melanoma cells from radial growth phase ( RGP ) to vertical growth phase ( VGP , metastatic phenotype ) are not yet well defined . We have demonstrated that the progression of human melanoma is associated with loss of expression of the transcription factor P05549 . In metastatic melanoma cells , this loss resulted in overexpression of P43121 / P43121 , P08253 , the thrombin receptor ( P25116 ) , and lack of c- P10721 expression . The transition from RGP to VGP is also associated with overexpression of the angiogenic factor P10145 . Additionally , the transition of melanoma cells from RGP to VGP is associated with overexpression of the transcription factors CREB and P39905 -1 , both of which may act as survival factors for human melanoma cells . Inactivation of CREB/ P39905 -1 activities in metastatic melanoma cells by dominant-negative CREB or by anti- P39905 -1 single chain antibody fragment ( ScFv ) , resulted in deregulation of P08253 and P43121 / P43121 , increased the sensitivity of melanoma cells to apoptosis , and inhibition of their tumorigenicity and metastatic potential in vivo . In this prospect article , we summarize our data on the role of P05549 and CREB/ P39905 -1 in the progression of human melanoma and report on the development of new fully human antibodies anti- P43121 / P43121 and anti- P10145 which could serve as new modalities for the treatment of melanoma . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Effects of cytokines on P15692 expression and secretion by human first trimester trophoblast cell line . PROBLEM : The mechanism through which vascular endothelial growth factor ( P15692 ) regulation occurs at the feto-maternal interface is poorly understood . The aim of this study was to investigate the effects of various cytokines on P15692 expression and secretion by trophoblast cells . METHOD OF STUDY : We investigated the effects of cytokines on P15692 expression in human first trimester trophoblast cell line by analyzing P15692 messenger RNA ( mRNA ) by reverse transcription-polymerase chain reaction and P15692 protein secretion by enzyme linked immunosorbent assay . RESULTS : The trophoblast cells expressed P15692 mRNA constitutively and the main subtypes were identified as VEGF121 and VEGF165 . When cultured in the presence of interferon ( IFN ) -gamma , interleukin ( IL ) - 1beta , tumor necrosis factor ( P01375 ) -alpha , P60568 , or P22301 , P15692 mRNA expression was found to be significantly increased by IL-1beta , P01579 and P01375 but to be unaffected by P60568 and P22301 . Moreover , P15692 secretion was most significantly increased by P01579 treatment . CONCLUSION : These results suggest that IL-1beta , P01579 , and P01375 may regulate the production of P15692 in early gestational trophoblasts . P39905 influences proliferation of osteoblastic cells . Little is known about the role of neurotrophic growth factors in bone metabolism . This study investigated the short-term effects of glial cell line-derived neurotrophic factor ( P39905 ) on calvarial-derived MC3T3-E1 osteoblasts . MC3T3-E1 expressed P39905 as well as its canonical receptors , GFRα1 and P07949 . Addition of recombinant P39905 to cultures in serum-containing medium modestly inhibited cell growth at high concentrations ; however , under serum-free culture conditions P39905 dose-dependently increased cell proliferation . P39905 effects on cell growth were inversely correlated with its effect on alkaline phosphatase ( AlP ) activity showing a significant dose-dependent inhibition of relative AlP activity with increasing concentrations of P39905 in serum-free culture medium . Live/dead and lactate dehydrogenase assays demonstrated that P39905 did not significantly affect cell death or survival under serum-containing and serum-free conditions . The effect of P39905 on cell growth was abolished in the presence of inhibitors to GFRα1 and P07949 indicating that P39905 stimulated calvarial osteoblasts via its canonical receptors . Finally , this study found that P39905 synergistically increased tumor necrosis factor-α ( P01375 -α ) -stimulated MC3T3-E1 cell growth suggesting that P39905 interacted with P01375 -α-induced signaling in osteoblastic cells . In conclusion , this study provides evidence for a direct , receptor-mediated effect of P39905 on osteoblasts highlighting a novel role for P39905 in bone physiology . P22681 mutation-related patterns of phosphorylation and sensitivity to tyrosine kinase inhibitors . Recurrent homozygous P22681 -inactivating mutations in myeloid malignancies decrease ubiquitin ligase activity that inactivates P12931 family kinases ( SFK ) and receptor tyrosine kinases ( RTK ) . However , the most important SFK and RTK affected by these mutations , and hence , the most important therapeutic targets , have not been clearly characterized . We compared SFK and RTK pathway activity and inhibitors in acute myeloid leukemia cell lines containing homozygous R420Q mutation ( GDM-1 ) , heterozygous deletion ( MOLM13 ) and wild-type ( WT ) P22681 ( THP1 , U937 ) . As expected with P22681 loss , GDM-1 displayed high P10721 expression and granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) hypersensitivity . Ectopic expression of WT P22681 decreased GDM-1 proliferation but not cell lines with WT P22681 . GDM-1 , but not the other cell lines , was highly sensitive to growth inhibition by dasatinib ( dual SFK and RTK inhibitor , LD50 50 nM ) ; there was less or no selective inhibition of GDM-1 growth by sunitinib ( RTK inhibitor ) , imatinib ( P00519 , P10721 inhibitor ) , or Q99463 ( SFK inhibitor ) . Phosphoprotein analysis identified phosphorylation targets uniquely inhibited by dasatinib treatment of GDM-1 , including a number of proteins in the P10721 and GM- P04141 receptor pathways ( for example , P10721 Tyr721 , P40763 Tyr705 ) . In conclusion , the promiscuous effects of P22681 loss on SFK and RTK signaling appear to be best targeted by dual SFK and RTK inhibition . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Creating a genotype-based dosing algorithm for acenocoumarol steady dose . DB01418 is a commonly prescribed anticoagulant drug for the prophylaxis and treatment of venous and arterial thromboembolic disorders in several countries . In counterpart of warfarin , there is scarce information about pharmacogenetic algorithms for steady acenocoumarol dose estimation . The aim of this study was to develop an algorithm of prediction for acenocoumarol.The algorithm was created using the data from 973 retrospectively selected anticoagulated patients and was validated in a second independent cohort adding up to 2,683 patients . The best regression model to predict stable dosage in the Primary Cohort included clinical factors ( age and body mass index , BSA ) and genetic variants ( Q9BQB6 , P11712 * and P78329 polymorphisms ) and explained up to 50 % of stable dose . In the validation study the clinical algorithm yielded an adjusted R²=0.15 ( estimation´s standard error=4.5 ) and the genetic approach improved the dose forecast up to 30 % ( estimation´s standard error=4.6 ) . Again , the best model combined clinical and genetic factors ( R² = 0.48 ; estimation´s standard error=4 ) which provided the best results of doses estimates within 20 % of the real dose in patients taking lower ( ≤ 7 mg/week ) or higher ( ≥ 25 mg/week ) acenocoumarol doses . In conclusion , we developed a prediction algorithm using clinical data and three polymorphisms in Q9BQB6 , P11712 * and P78329 genes that provided a steady acenocoumarol dose for about 50 % of patients in the Validation Cohort . Such algorithm was especially useful to patients who need higher or lower acenocoumarol doses , those patients with higher time required until their stabilisation and are more prone to suffer a treatment derived complication . Differential regulation of P14780 and P16035 expression in malignant melanoma developed in metallothionein/ P07949 transgenic mice . We recently established a metallothionein-I(MT)/ P07949 transgenic mouse line in which skin melanosis , benign melanocytic tumor and malignant melanoma develop stepwise . Malignant melanoma cells but not benign melanocytic tumor cells had metastatic ability in transgenic mice . In the present study , we investigated the expression of several matrix metalloproteinases ( MMPs ) and tissue inhibitors of matrix metalloproteinases ( TIMPs ) , including P03956 , P08253 , P08254 , P09237 , P14780 , P50281 , P01033 and P16035 , in these tumors . Western and northern blot analyses revealed that malignant transformation of melanocytic tumors developed in MT/ P07949 transgenic mice accompanied with upregulation of P14780 and downregulation of P16035 . Expression of other MMP and P01033 genes examined was very low or undetectable in both benign and malignant tumors . Since activation of P14780 in malignant tumors was detected by gelatin zymography , these results suggest that imbalance of expression of the P14780 and P16035 genes might be associated with metastatic ability of melanoma cells developed in MT/ P07949 transgenic mice . DB09078 : first global approval . DB09078 ( Lenvima™ ) is a multitargeted receptor kinase inhibitor that inhibits the kinase activities of vascular endothelial-derived growth factor receptors 1 , 2 and 3 , fibroblast growth factor receptors 1 , 2 , 3 and 4 , platelet-derived growth factor receptor α , P07949 and P10721 . In addition to their role in normal cellular function , these kinases have been implicated in pathogenic angiogenesis , tumour growth and cancer progression . DB09078 is being developed by Eisai Co . Ltd for the treatment of solid tumours , primarily for differentiated thyroid cancer , and other malignancies . A capsule formulation of the drug has received approval in the USA for use in locally recurrent or metastatic , progressive , radioactive iodine-refractory differentiated thyroid cancer . DB09078 is in pre-registration for this indication in the EU , Australia , Brazil , Canada , Japan , South Korea , Russia , Singapore and Switzerland , and is in phase 3 development in Argentina , Chile and Thailand . DB09078 has orphan designation in the EU and Japan for use in differentiated thyroid cancer . In addition , an ongoing global , phase 3 trial is evaluating the use of lenvatinib as first-line treatment in unresectable hepatocellular carcinoma . This article summarizes the milestones in the development of lenvatinib leading to this first approval in locally recurrent or metastatic , progressive , radioactive iodine-refractory differentiated thyroid cancer . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . P25116 genotype influences platelet aggregation and procoagulant responses in patients with coronary artery disease prior to and during clopidogrel therapy . Genetic variations of the protease-activated receptor-1 ( P25116 ) have been associated with platelet receptor density and linked to thrombin receptor-activating peptide ( TRAP ) -induced phenotypes of platelet aggregation and P16109 expression . We investigated whether the P25116 intervening sequence-14 A > T dimorphism influences platelet procoagulant activity . We also determined whether the Q9H244 antagonist clopidogrel could offset any observed functional polymorphism of the P25116 receptor by inhibiting Q9H244 -mediated amplification of TRAP-induced responses . We studied 54 patients listed for elective percutaneous coronary intervention assessing TRAP-induced platelet aggregation and markers of procoagulant activity . Platelet responses were measured at baseline , 4 h post clopidogrel 300 mg , and 10 and 28 days following clopidogrel 75 mg daily . Each patient was genotyped for the P25116 intervening sequence-14 A/T dimorphism . Increased platelet aggregation and procoagulant responses were observed with P25116 A allele homozygotes . DB00758 significantly inhibited these platelet responses regardless of P25116 genotype , but did not offset the hyper-reactivity associated with the A/A homozygotes . We conclude that a common sequence variation within the P25116 gene influences TRAP-induced platelet procoagulant activity as well as aggregation . Higher platelet reactivity associated with P25116 IVSn-14 A allele homozygotes persists despite clopidogrel therapy . These individuals may be at higher risk of thromboembolic events and may require additional anti-platelet medication . Putative role of HIF transcriptional activity in melanocytes and melanoma biology . Hypoxia-inducible factor-1α ( HIF-1α ) is a highly oxygen sensitive bHLH protein that is part of the heterodimeric Q9BYW2 transcription factor . Under hypoxic stress , Q9BYW2 activity is induced to control expression of multiple downstream target genes , including vascular endothelial growth factor ( P15692 ) . The normal epidermis exists in a constant mild hypoxic microenvironment and constitutively expresses HIF-1α and HIF-2α . Expression of HIF-1α and/or HIF-2α has been suggested to correlate with the increased malignant potential of melanocytes , therefore , failures of melanoma therapies may be partially linked to high HIF activity . Notably , melanomas that have the V600E P15056 mutation exhibit increased HIF-1α expression . We have utilized a bioinformatics approach to identify putative hypoxia response elements ( HREs ) in a set of genes known to participate in the process of melanogenesis ( includingTRPM1 , Q9UMX9 , P01112 , C- P10721 , P40967 and P06850 ) . While some of the mechanistic links between these genes and the HIF pathway have been previously explored , others await further investigation . Although agents targeting HIF activity have been proposed as novel treatment modalities for melanoma , there are currently no clinical trials in progress to test their efficacy in melanoma . Vascular endothelial growth factor receptors in osteoclast differentiation and function . Osteoclasts are derived from haematopoietic stem cell precursors of the monocyte/macrophage cell lineage , through interaction with factors that are believed to include P09603 and O14788 . P15692 is a proangiogenic cytokine that has been shown to promote osteoclast differentiation and survival . In this study , we assessed the role of P15692 and its receptors in osteoclastogenesis , in vitro , by culturing osteoclast precursors in the presence of P15692 , P15692 receptor-specific ligands , and blocking antibodies to P15692 receptors . Activation of P17948 in the presence of O14788 induces osteoclast differentiation . Stimulating the receptors individually induced increased resorption by osteoclasts compared to controls but not to the level observed when stimulating both receptors simultaneously . We have shown that P15692 induces osteoclast differentiation through its action on P17948 . The way in which P15692 mediates its effect on mature osteoclast activity , however , may be through its interaction with both receptor subtypes . Sustained vascular endothelial growth factor delivery enhances angiogenesis and perfusion in ischemic hind limb . PURPOSE : We hypothesized that sustained delivery of vascular endothelial growth factor ( P15692 ) using a polymer [ 85:15 poly(lactide-co-glycolide) ( P00747 ) ] would enhance angiogenesis and improve perfusion of ischemic tissue . METHODS : C57BL/6J mice ( n = 20/group ) underwent unilateral hind limb ischemia surgery and were randomized to groups of no scaffold implantation ( 0-Implant ) , unloaded scaffold implantation ( Empty- P00747 ) , or implantation of scaffolds incorporating 3 microg of VEGF165 ( P00747 - P15692 ) . Endpoints included laser Doppler perfusion imaging ( LDPI , ischemic/nonischemic limb , % ) , local vessel counts , immunohistochemistry for CD31 , and alpha-smooth muscle actin . In vitro release kinetics of P15692 from P00747 was also measured . RESULTS : P00747 - P15692 resulted in improved lower extremity perfusion vs. controls as measured by LDPI % at 7 , 14 , 21 , and 28 days ( p < 0.05 ) . P00747 - P15692 was associated with significantly greater percentage of vessels staining for CD31 and alpha-smooth muscle actin compared to the Empty- P00747 or 0-Implant ( p < 0.05 for both ) . CONCLUSIONS : The P00747 - P15692 scaffolds resulted in sustained P15692 delivery , improved tissue perfusion , greater capillary density , and more mature vasculature compared to the controls . The sustained-release P00747 polymer vehicle is a promising delivery system for therapeutic neovascularization applications . 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 . Membrane P48023 kills human peripheral blood T lymphocytes , and soluble P48023 blocks the killing . It has been believed that the Fas expressed on human peripheral blood T cells ( P10721 ) is nonfunctional , because these cells are insensitive to agonistic anti-Fas/Apo-1 mAbs that efficiently kill in vitro-activated T cells and many Fas-expressing cell lines . Here , we demonstrate that membrane-bound P48023 ( P48023 ) kills both fresh and in vitro-activated P10721 , indicating that the Fas expressed on fresh P10721 is functional . In contrast , soluble P48023 kills only the latter . Naive T cells in umbilical cord blood do not express Fas , but can be induced to express Fas by P01579 or by a combination of P60568 and anti- P10747 mAb , after which they acquire sensitivity to membrane but not to soluble P48023 . Soluble P48023 inhibited the killing of fresh P10721 by membrane P48023 . These results indicate that the shedding of P48023 from the membrane is a mechanism for downregulating at least part of its killing activity . Temporal and pharmacological characterization of angiostatin release and generation by human platelets : implications for endothelial cell migration . Platelets play an important role in thrombosis and in neo-vascularisation as they release and produce factors that both promote and suppress angiogenesis . Amongst these factors is the angiogenesis inhibitor angiostatin , which is released during thrombus formation . The impact of anti-thrombotic agents and the kinetics of platelet angiostatin release are unknown . Hence , our objectives were to characterize platelet angiostatin release temporally and pharmacologically and to determine how angiostatin release influences endothelial cell migration , an early stage of angiogenesis . We hypothesized anti-platelet agents would suppress angiostatin release but not generation by platelets . Human platelets were aggregated and temporal angiostatin release was compared to vascular endothelial growth factor ( P15692 ) . Immuno-gold electron microscopy and immunofluorescence microscopy identified α-granules as storage organelles of platelet angiostatin . Acetylsalicylic acid , MRS2395 , P08514 /IIIa blocking peptide , and aprotinin were used to characterize platelet angiostatin release and generation . An endothelial cell migration assay was performed under hypoxic conditions to determine the effects of pharmacological platelet and angiostatin inhibition . Compared to P15692 , angiostatin generation and release from α-granules occurred later temporally during platelet aggregation . Consequently , collagen-activated platelet releasates stimulated endothelial cell migration more potently than maximally-aggregated platelets . Platelet inhibitors prostacyclin , S-nitroso-glutathione , acetylsalicylic acid , and P08514 /IIIa blocking peptide , but not a Q9H244 inhibitor , suppressed angiostatin release but not generation . Suppression of angiostatin generation in the presence of acetylsalicylic acid enhanced platelet-stimulated endothelial migration . Hence , the temporal and pharmacological modulation of platelet angiostatin release may have significant consequences for neo-vascularization following thrombus formation . Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries . BACKGROUND : Atherosclerosis affects aorta , coronary , carotid , and iliac arteries most frequently than any other body vessel . There may be common molecular pathways sustaining this process . Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis . RESULTS : We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression . We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis . Caveolae and JAK/ P35610 pathways , and P06702 / P05109 interacting proteins are certainly involved in the development of vascular disease . We found that the system of caveolae is directly connected with genes that respond to hormone receptors , and indirectly with the apoptosis pathway . Cytokines , chemokines and growth factors released in the blood flux were investigated in parallel . High levels of RANTES , IL-1ra , MIP-1 alpha , MIP-1 beta , P60568 , P05112 , P05113 , P05231 , P13232 , Q16552 , DB00102 , P15692 and P01579 were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels . CONCLUSION : The pattern of cytokine and P06702 / P05109 up-regulation characterizes atherosclerosis as a proinflammatory disorder . Activation of the JAK/ P35610 pathway is confirmed by the up-regulation of P05231 , P42224 , Q00978 and Q13651 genes in coronary and carotid plaques . The functional network constructed in our research is an evidence of the central role of P35610 protein and the caveolae system to contribute to preserve the plaque . Moreover , Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype . In vitro transforming potential , intracellular signaling properties , and sensitivity to a kinase inhibitor ( sorafenib ) of P07949 proto-oncogene variants Glu511Lys , Ser649Leu , and Arg886Trp . Multiple endocrine neoplasia type 2 and a subset of apparently sporadic medullary thyroid carcinoma ( AS- P04629 ) are caused by germ line activating point mutations of the rearranged during transfection ( P07949 ) proto-oncogene . P07949 encodes a receptor with tyrosine kinase activity that targets several intracellular signaling cascades , such as DB01367 -RAF- P27361 /2 , PIK3-AKT , and P35610 transcription factors . The objective of this study was to assess the function of three germ line P07949 variants Arg886Trp , Ser649Leu , and Glu511Lys of undetermined pathogenic significance , which were found in three kindreds of isolated AS- P04629 . For this purpose , we employed vectors expressing each of the P07949 variants and measured the number of NIH3T3 transformation foci and soft agar colonies , the degree of activation of known P07949 intracellular signaling targets ( P27361 /2 , P42224 , P40763 , and TCF4 ) , and the extent of P27361 /2 inhibition on sorafenib treatment . We found that P07949 variants Arg886Trp and Glu511Lys have shown increased in vitro transforming potential in a glial-derived neurotrophic factor-dependent manner . In contrast , the Ser649Leu variant did not significantly increased the number of foci and agar colonies relative to wild-type P07949 ( P07949 -WT ) . The variants Glu511Lys and Arg886Trp showed 10- and 12.5-fold P27361 /2 activation respectively , that was significantly higher than that observed for P07949 -WT ( fivefold ) . Increased levels of P42224 and TCF4 activation were only observed for P07949 Arg886Trp ( 2.5- and 3-fold versus 1.2- and 2-fold in P07949 -WT respectively ) . The three P07949 variants analyzed here were sensitive to treatment with sorafenib . In conclusion , our results allow to classify previously uncharacterized P07949 genotypes , which may be of use to define follow-up and therapeutic regimens . Amniotic DB05914 with robust chemotactic properties are effective in the treatment of a myocardial infarction model . BACKGROUND : We previously reported that amniotic DB05914 ( AMMs ) possess high angio-vasulogenic properties . In this study , we investigated the chemotactic abilities of AMMs for improved cardiac function and regenerative angiogenesis . METHODS : The expressions of chemotactic and angiogenic genes were determined by qRT-PCR . Myocardial infarction ( MI ) was induced in NOD/SCID mice and cells were directly transplanted into the border regions of ischemic heart tissue . Immunohistochemical analysis was also conducted . RESULTS : AMMs significantly expressed the representative chemotactic factor P04001 -2 , Q99733 as well as angiogenic factor Hif-1a . AMMs also highly expressed the chemokine receptors P41597 , P51677 and P51681 . AMM transplantation improved left ventricular function , capillary density , angiogenic cytokine levels , angiopoetin ( Ang ) -1 and vascular endothelial growth factor ( P15692 ) levels in affected tissue . Immunohistochemical assaying also revealed increased engraftment and endothelial phenotypes . CONCLUSION : Our findings suggest that due to elevated survival and related chemotactic potential , AMMs are a promising stem cell source for the treatment of ischemic cardiovascular disease . Effects of ellagic Acid on angiogenic factors in prostate cancer cells . BACKGROUND : Several natural antioxidants , including ellagic acid ( EA ) , have been reported to have chemotherapeutic activity in vivo and in vitro settings . Cytochrome P450 ( CYP ) activity and synthesis of both epoxyeicosatrienoic acids ( EETs ) and 20-hydroxy-5,8,11,14-eicosatetraenoic acid ( 20-HETE ) , together with vascular endothelial growth factor ( P15692 ) and heme oxygenase system ( HO ) have emerged as important modulators of tumor growth and metastasis . METHODS : The anti-angiogenic effects of EA were investigated in the human prostatic cancer cell line LnCap . P09601 , P30519 , P51589 and soluble epoxyde hydrolase ( sEH ) expressions were evaluated by western blotting . Levels of P15692 and osteoprotegerin ( O00300 ) were determined in the culture supernatant using an ELISA assay , while CYP mRNAs were determined by qRT-PCR . RESULTS : EA treatment induced a significant decrease ( p < 0.05 ) in P09601 , P30519 and P51589 expression , and in P15692 and O00300 levels . Similarly P51589 , P78329 and CYPA22 mRNAs were significantly ( p < 0.05 ) down-regulated by EA treatment . The decrease in P51589 mRNA was associated with an increase in sEH expression . CONCLUSIONS : RESULTS reported in the present study highlighted the ability of EA to modulate a new pathway , in addition to anti-proliferative and pro-differentiation properties , via a mechanism that involves a decrease in eicosanoid synthesis and a down-regulation of the HO system in prostate cancer . 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 . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . Potential cellular signatures of viral infections in human hematopoietic cells . Expression profiling of cellular genes was performed using a 10,000 cDNA human gene array in order to identify expression changes following chronic infection of human hematopoietic cells with Kaposi 's Sarcoma-associated Virus ( KSHV ) also known as Human Herpesvirus 8 ( HHV8 ) and Human T cell leukemia virus-1 ( HTLV-1 ) . We performed cell-free in vitro infection of primary bone marrow derived P28906 + cells using semi-purified HHV8 and a mature P60568 dependent T cell line , P10721 225 , using highly concentrated viral stocks prepared from an infectious molecular clone of HTLV-1 . Thirty days post infection , mRNA was isolated from infected cultures and uninfected controls and submitted for microarray analysis . More than 400 genes were differentially expressed more than two-fold following HHV8 infection of primary bone marrow derived P28906 + cells . Of these 400 , interferon regulatory factor 4 ( Q15306 ) , cyclin B2 , P20226 -associated factor , eukaryotic elongation factor and pim 2 were up-regulated more than 3.5 fold . In contrast , less than 100 genes were differentially expressed more than two-fold following chronic infection of a mature T cell line with HTLV-1 . Of these , only cdc7 was up-regulated more than 3.5 fold . These data may provide insight into cellular signatures of infection useful for diagnosis of infection as well as potential targets for therapeutic intervention . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . 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 . The role of vascular endothelial growth factor and matrix metalloproteinases in canine lymphoma : in vivo and in vitro study . BACKGROUND : Canine lymphoma represents the most frequent haematopoietic cancer and it shares some similarities with human non-Hodgkin lymphoma . Matrix metalloproteinases ( MMPs ) and vascular endothelial growth factor ( P15692 ) play a coordinated role during invasion and proliferation of malignant cells ; however , little is known about their role in canine haematologic malignancies . The aim of this study was to investigate the mRNA and protein expression of P15692 and the most relevant MMPs in canine lymphoma . Lymph node aspirates from 26 B-cell and 21 T-cell lymphomas were collected . The protein expression levels of P14780 , P08253 and P15692 were evaluated by immunocytochemistry , and the mRNA levels of P08253 , P14780 , P50281 , P01033 , P16035 , O95980 , P15692 and P15692 -164 were measured using quantitative RT-PCR . RESULTS : P50281 , P01033 and O95980 mRNA levels were significantly higher in T-cell lymphomas than in B-cell lymphomas . Higher mRNA and protein levels of P14780 and P15692 were observed in T-cell lymphomas than in B-cell lymphomas and healthy control lymph nodes . A positive correlation was found between P14780 and P15692 in T-cell lymphomas . Moreover , P14780 , P50281 , P01033 and P15692 were expressed at the highest levels in high-grade T-cell lymphomas . CONCLUSIONS : This study provides new information on the expression of different MMPs and P15692 in canine lymphoma , suggesting a possible correlation between different MMPs and P15692 , immunophenotype and prognosis .

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New insights into the role of calcium-sensing receptor activation . The discovery of the calcium-sensing receptor ( P41180 ) prompted the identification of substances that affect its function . DB01012 , for example , is a drug that allosterically modifies the receptor so as to increase its sensitivity to circulating calcium ( thus the name " calcimimetic " ) and in this way decreases parathyroid hormone secretion . Clinical use of cinacalcet is already approved for the treatment of primary and secondary hyperparathyroidism , but research is ongoing to identify further potential actions of this drug . The effects and functions of the P41180 have been evaluated in different systems and tissues , beyond parathyroid glands , such arterial walls . A complete understanding of the properties of calcimimetics are of obvious clinical interest , since therapeutic indications may be affected accordingly . Vitamin D increases plasma renin activity independently of plasma Ca2+ via hypovolemia and β-adrenergic activity . 1 , 25-Dihydroxycholechalciferol ( calcitriol ) and 19-nor-1 , 25-dihydroxyvitamin D2 ( paricalcitol ) are vitamin D receptor ( P11473 ) agonists . Previous data suggest P11473 agonists may actually increase renin-angiotensin activity , and this has always been assumed to be mediated by hypercalcemia . We hypothesized that calcitriol and paricalcitol would increase plasma renin activity ( P06703 ) independently of plasma Ca(2+) via hypercalciuria-mediated polyuria , hypovolemia , and subsequent increased β-adrenergic sympathetic activity . We found that both calcitriol and paricalcitol increased P06703 threefold ( P < 0.01 ) . Calcitriol caused hypercalcemia , but paricalcitol did not . Both calcitriol and paricalcitol caused hypercalciuria ( 9- and 7-fold vs. control , P < 0.01 ) and polyuria ( increasing 2.6- and 2.2-fold vs. control , P < 0.01 ) . Paricalcitol increased renal calcium-sensing receptor ( P41180 ) expression , suggesting a potential cause of paricalcitol-mediated hypercalciuria and polyuria . Volume replacement completely normalized calcitriol-stimulated P06703 and lowered plasma epinephrine by 43 % ( P < 0.05 ) . β-Adrenergic blockade also normalized calcitriol-stimulated P06703 . P35354 inhibition had no effect on calcitriol-stimulated P06703 . Our data demonstrate that vitamin D increases P06703 independently of plasma Ca(2+) via hypercalciuria , polyuria , hypovolemia , and increased β-adrenergic activity . Behind the curtain : cellular mechanisms for allosteric modulation of calcium-sensing receptors . DB01373 -sensing receptors ( P41180 ) are integral to regulation of systemic Ca(2+) homeostasis . Altered expression levels or mutations in P41180 cause Ca(2+) handling diseases . P41180 is regulated by both endogenous allosteric modulators and allosteric drugs , including the first Food and Drug Administration-approved allosteric agonist , DB01012 HCl ( Sensipar® ) . Recent studies suggest that allosteric modulators not only alter function of plasma membrane-localized P41180 , but regulate P41180 stability at the endoplasmic reticulum . This brief review summarizes our current understanding of the role of membrane-permeant allosteric agonists in cotranslational stabilization of P41180 , and highlights additional , indirect , signalling-dependent role(s) for membrane-impermeant allosteric drugs . Overall , these studies suggest that allosteric drugs act at multiple cellular organelles to control receptor abundance and hence function , and that drug hydrophobicity can bias the relative contributions of plasma membrane and intracellular organelles to P41180 abundance and signalling . Growth inhibition of human glioma cells by transfection-induced P21 and its effects on telomerase activity . The aim of this study is to investigate the effect of the P38936 gene transfection on the growth of cultured human glioma cell lines , and analyze the telomerase activity , and detection of telomerase components in P38936 transfectant . The P38936 gene was transfected into human glioma cell lines , U251MG and T98G with our novel liposome . The cell growth was assessed by counting the number of trypan blue-excluding cells in a hemocytometer and flow cytometry analysis . The expression of P21 protein and its mRNA were examined by Western and Northern blot analysis . The telomerase activity was assayed by TRAP ( telomerase repeat amplification protocol ) /TRAP- Q9Y251 ( hybridization protection assay ) method qualitatively and quantitatively . The length of telomere was measured by Southern blot analysis . The expression of telomerase components ( hTERT , hTERC and TEP1 ) were examined by RT-PCR ( reverse transcriptase-polymerase chain reaction ) . The P38936 transfectant demonstrated the expression of P21 protein and its mRNA . The P38936 transfection of human glioma cells results in growth inhibition and G0/ P55008 arrest . The P38936 transfectant revealed a decrease of telomerase activity and hTERT expression as compared with control cells . These results suggest that P38936 transfection induces G0/ P55008 arrest in human glioma cells which associates with the reduction in the telomerase activity and hTERT expression . [ DB01012 -- a new drug for the treatment of secondary hyperparathyroidism in patients with uraemia , parathyroid cancer or primary hyperparathyroidism ] . DB01012 is a new drug with antiparathyroid effects that belongs to the class of calcimimetics . It increases the sensitivity of the calcium-sensing receptor ( P41180 ) to calcium , thus inducing a decrease in plasma parathyroid ( PTH ) levels . In patients with uncontrolled secondary hyperparathyroidism due to uremia , cinacalcet has been shown to decrease the levels of PTH even in those optimally treated with calcium and 1-ahydroxylated vitamin D . DB01012 decreases plasma calcium and plasma PTH levels in patients with primary hyperparathyroidism or parathyroid cancer . Successful use of bisphosphonate and calcimimetic in neonatal severe primary hyperparathyroidism . Neonatal primary hyperparathyroidism ( NPHT ) is associated with an inactivating homozygous mutation of the calcium sensing receptor ( P41180 ) . The P41180 is expressed most abundantly in the parathyroid glands and the kidney and regulates calcium homeostasis through its ability to modulate parathormone secretion and renal calcium reabsorption . NPHT leads to life threatening hypercalcemia , nephrocalcinosis , bone demineralization , and neurologic disabilities . Surgery is the treatment of choice . While waiting for surgery , bisphosphonates offer a good alternative to deal with hypercalcemia . DB01012 is a class II calcimimetic that increases P41180 affinity for calcium , leading to parathormone suppression and increased calcium renal excretion . At present , there is little evidence as to whether cinacalcet could improve the function of mutant P41180 in NPHT . We report a case of NPHT , treated successfully with bisphosphonates and cinacalcet after surgery failure . To our knowledge , it is the first time cinacalcet has been used for NPHT . Signalling pathways involved in retinal endothelial cell proliferation induced by advanced glycation end products : inhibitory effect of gliclazide . AIM : We have previously demonstrated that advanced glycation end products ( AGEs ) stimulate bovine retinal endothelial cell ( BREC ) proliferation through induction of vascular endothelial growth factor ( P15692 ) production by these cells . We have also shown that gliclazide , a sulfonylurea which decreases oxidative stress , inhibits this effect . The aim of the present study was to characterize the signalling pathways involved in P51606 -induced BREC proliferation and P15692 production and mediating the inhibitory effect of gliclazide on these biological events . METHODS : BRECs were treated or not treated with AGEs in the presence or absence of gliclazide , antioxidants , protein kinase C ( PKC ) , mitogen-activated protein kinase ( MAPK ) or nuclear factor-kappaB ( NF-kappaB ) inhibitors . BREC proliferation was assessed by measuring [ 3H ] -thymidine incorporation into DNA . Activation of PKC , MAPK and NF-kappaB signal transduction pathways and determination of P15692 expression were assessed by Western blot analysis using specific antibodies . MAPK activity was also determined by an in vitro kinase assay . RESULTS : Treatment of BRECs with AGEs significantly increased cell proliferation and P15692 expression . AGEs induced P05771 translocation , extracellular signal-regulated protein kinase 1/2 and NF-kappaB activation in these cells . Pharmacological inhibition of these signalling pathways abolished P51606 effects on cell proliferation and P15692 expression . Exposure of BRECs to gliclazide or antioxidants such as vitamin E or N-acetyl-l-cysteine resulted in a significant decrease in P51606 -induced activation of PKC- , MAPK- and NF-kappaB-signalling pathways . CONCLUSIONS : Our results demonstrate the involvement of PKC , MAPK and NF-kappaB in P51606 -induced BREC proliferation and P15692 expression . DB01120 inhibits BREC proliferation by interfering with these intracellular signal transduction pathways . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . P00734 kringle-2 induces death of mesencephalic dopaminergic neurons in vivo and in vitro via microglial activation . We have shown that prothrombin kringle-2 ( pKr-2 ) , a domain of human prothrombin distinct from thrombin could activate cultured rat brain microglia in vitro . However , little is known whether pKr-2-induced microglial activation could cause neurotoxicity on dopaminergic ( DA ) neurons in vivo . To address this question , pKr-2 was injected into the rat substantia nigra ( SN ) . Tyrosine hydroxylase ( TH ) immunohistochemistry experiments demonstrate significant loss of DA neurons seven days after injection of pKr-2 . In parallel , pKr-2-activated microglia were detected in the SN with OX-42 and OX-6 immunohistochemistry . Reverse transcription PCR and double-label immunohistochemistry revealed that activated microglia in vivo exhibit early and transient expression of inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-2 ( P35354 ) and several proinflammatory cytokines . The pKr-2-induced loss of SN DA neurons was partially inhibited by the NOS inhibitor N(G)-nitro-L-arginine methyl ester hydrochloride , and the P35354 inhibitor DuP-697 . P27361 /2 , c-Jun N-terminal kinase and p38 mitogen-activated protein kinase were activated in the SN as early as 1 hr after pKr-2 injection , and localized within microglia . Inhibition of these kinases led to attenuation of mRNA expression of P35228 , P35354 and several proinflammatory cytokines , and rescue of DA neurons in the SN . Intriguingly , following treatment with pKr-2 in vitro , neurotoxicity was detected exclusively in co-cultures of mesencephalic neurons and microglia , but not microglia-free neuron-enriched mesencephalic cultures , indicating that microglia are required for pKr-2 neurotoxicity . Our results strongly suggest that microglia activated by endogenous compound(s) , such as pKr-2 , are implicated in the DA neuronal cell death in the SN . Recombinant human prothrombin kringle-2 induces bovine capillary endothelial cell cycle arrest at G0- P55008 phase through inhibition of cyclin D1/ P11802 complex : modulation of reactive oxygen species generation and up-regulation of cyclin-dependent kinase inhibitors . P00734 is a plasma glycoprotein involved in blood coagulation and , as we have previously reported , prothrombin kringles inhibit BCE ( bovine capillary endothelial ) cell proliferation . To reveal the mechanism , we investigated the influence of rk-2 ( recombinant human prothrombin kringle-2 ) on the BCE cell cycle progression and ROS ( reactive oxygen species ) generation using FACS ( fluorescence-activated cell sorter ) analysis . Cell cycle analysis showed a decrease of G(1) phase cells in cells treated with P09038 ( basic fibroblast growth factor ) and an increase in cells treated with rk-2 , as compared with the control cells . But , the portion of the S phase was reversed . In Western blot analysis , P09038 induced cytoplasmic translocation of P38936 (Waf1/Cip1) and p27(Kip1) and phosphorylation of p27(Kip1) but rk-2 treatment inhibited translocation of P38936 (Waf1/Cip1) and p27(Kip1) from nucleus to cytoplasm and phosphorylation of p27(Kip1) . Also , rk-2 induced up-regulation of p53 and nuclear P38936 (Waf1/Cip1) and inhibited the cyclin D1/ P11802 ( cyclin-dependent kinase 4 ) complex . The ROS level of rk-2-treated BCE cells was increased 2-fold when compared with the control , but treatment with Q9C000 ( N-Acetyl-L : -cysteine ) , an anti-oxidant , decreased ROS generation about 55 % as compared with the rk-2 treatment . Q9C000 treatment also restored cell cycle progression inhibited by rk-2 and down-regulated p53 and nuclear P38936 (Waf1/Cip1) expression induced by rk-2.These data suggest that rk-2 induces the BCE cell cycle arrest at G(0)-G(1) phase through inhibition of the cyclin D1/ P11802 complex caused by increase of ROS generation and nuclear cyclin-dependent kinase inhibitors . Discovery of a calcimimetic with differential effects on parathyroid hormone and calcitonin secretion . Calcimimetics are positive allosteric modulators to the calcium-sensing receptor ( P41180 ) . Activation of the P41180 inhibits the secretion of parathyroid hormone ( PTH ) , stimulates the secretion of calcitonin , and decreases serum calcium ( Ca(2+) ) . DB01012 , a second-generation calcimimetic , is used therapeutically to control PTH in patients with chronic kidney disease who are on dialysis with secondary hyperparathyroidism . A calcimimetic that displays increased separation of PTH versus Ca(2+) lowering in patients would potentially allow the use of calcimimetics to treat patients in earlier stages of renal disease because hypocalcemia can develop in this population . Toward this end , we developed a third-generation calcimimetic , determined the molecular pharmacological properties of it using an operation model of allosteric modulation/agonism , and measured the compound effects on PTH , serum ionized Ca(2+) , and calcitonin levels in 5/6 nephrectomized rats . We found the new molecule effectively reduced PTH levels without promoting calcitonin secretion or hypocalcemia . Furthermore , our third-generation molecule was less efficacious at promoting calcitonin secretion from human thyroid carcinoma cells compared with 3-(2-chlorophenyl)-N- ( ( 1R ) -1-(3-methoxyphenyl)ethyl ) -1-propanamine ( R-568 ) , a first-generation calcimimetic . These data provide evidence that calcimimetics with increased potency can be used to lower PTH without production of significant hypocalcemia because the threshold for inhibition of PTH secretion is much lower than the threshold for calcitonin secretion . [ Role of Q02750 / P27361 , 2 pathways in the calcium-sensing receptor mediation of hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells ] . OBJECTIVE : To explore the cell signal transduction pathway of calcium-sensing receptor ( P41180 ) mediated hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells ( PASMCs ) . METHODS : The expressions of proliferating cell nuclear antigen ( P12004 ) and extracellular signal-regulated protein kinase 1 , 2 ( P27361 , 2 ) were analyzed by Western blot . Cell proliferation was tested by a 5-bromo-2-deoxyuridine ( BrdU ) incorporation assay . Cell cycle and proliferation index ( PI ) were analyzed by flow cytometry . RESULTS : Hypoxia significantly increased the expression of P12004 ( 0.528 ± 0.028 ) , p- P27361 , 2 ( 1.12 ± 0.05 , 0.91 ± 0.06 ) , BrdU incorporation ( 143.3 ± 4.2 ) and cell proliferation index ( 12.5 ± 0.9 ) ( all P < 0.05 , versus control group , 0.243 ± 0.025 , 0.47 ± 0.03 , 0.40 ± 0.03 , 100.0 ± 5.4 , 7.5 ± 1.2 ) . Gadolinium chloride ( GdCl3 , a P41180 agonist ) amplified the effect of hypoxia ( 0.770 ± 0.039 , 1.50 ± 0.06 , 1.61 ± 0.05 , 187.4 ± 3.9 , 19.8 ± 0.6 , all P < 0.05 ) . PD98059 ( a Q02750 inhibitor ) decreased the up-regulation of P12004 expression , BrdU incorporation and the increase of cell proliferation index induced by hypoxia and GdCl3 in PASMCs ( 0.441 ± 0.020 , 0.71 ± 0.07 , 0.72 ± 0.06 , 115.5 ± 4.0 , 9.3 ± 1.1 , all P < 0.05 ) . CONCLUSION : DB01373 -sensing receptor mediates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells through P27361 , 2 pathways . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . Persistent Cdk2 inactivation drives growth arrest of P11274 - P00519 -expressing cells in response to dual inhibitor of P12931 and P00519 kinases SKI606 . Complementary inhibition of tyrosine and P12931 kinases implement dual P12931 / P00519 inhibitor effects in chronic myeloid leukemia ( CML ) . Here , we show that one such inhibitor , DB06616 , induces persistent Cdk2 inactivation leading to growth arrest of P11274 - P00519 -expressing cells either IM-sensitive or driven to IM-resistance by other events than gene overexpression and point mutations . Inhibition of Akt serine/threonine kinase , a phosphatidylinositol 3 kinase ( PI-3k ) target that integrates Q92817 TK signaling with membrane-associated P12931 kinases , is a central component of restored expression and subcellular redistribution of Cdk2 regulatory signals ( P38936 and p27 and Cdc25A phosphatase ) in response to DB06616 . The putative roles of growth factor ( namely P08700 ) autocrine loop in P11274 - P00519 -expressing progenitor progression towards a drug-resistant phenotype are discussed . DB01373 -sensing receptor activation in chronic kidney disease : effects beyond parathyroid hormone control . Secondary hyperparathyroidism ( SHPT ) is an important complication of advanced chronic kidney disease ( CKD ) . DB01012 , an allosteric modulator of the calcium-sensing receptor ( P41180 ) expressed in parathyroid glands , is the only calcimimetic approved to treat SHPT in patients on dialysis . By enhancing P41180 sensitivity for plasma extracellular calcium ( Ca(2+)0 ) , cinacalcet reduces serum parathyroid hormone , Ca(2+)0 , and serum inorganic phosphorous concentrations , allowing better control of SHPT and CKD-mineral and bone disorders . Of interest , the P41180 also is expressed in a variety of tissues where its activation regulates diverse cellular processes , including secretion , apoptosis , and proliferation . Thus , the existence of potential off-target effects of cinacalcet can not be neglected . This review summarizes our current knowledge concerning the potential role(s) of the P41180 expressed in various tissues in CKD-related disorders , independently of parathyroid hormone control . Association between severe toxicity of nilotinib and P22309 polymorphisms in Japanese patients with chronic myelogenous leukemia . BACKGROUND : DB04868 is a P11274 - P00519 kinase inhibitor approved for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia ( CML ) . The P22309 ( P22309 ) polymorphism P22309 *28 ( *28 ) /*28 has been linked to an increased risk of hyperbilirubinemia in patients with CML who receive nilotinib . Beside *28 , P22309 *6 ( *6 ) is another important variant allele in Japanese patients because it is associated with adverse events of irinotecan , metabolized by P22309 . We retrospectively investigated the association between severe toxicity of nilotinib and P22309 polymorphisms ( *6 and*28 ) in Japanese patients with CML . PATIENTS AND METHODS : Eight patients with cytogenetically confirmed CML who were receiving nilotinib were studied to explore the association of P22309 polymorphisms with severe nilotinib-related toxicity . Genotyping analyses were determined for *6 and *28 . RESULTS : All 3 patients with the *6/*6 or *6/*28 genotype had severe toxicity , including QT interval prolongation ( grade 3 ) , elevated lipase levels ( grade 3 ) plus hyperbilirubinemia ( grade 2 ) , and anemia ( grade 3 ) plus hepatic cyst hemorrhage ( grade 2 ) in 1 patient each . Among the 5 patients with the *6/*1 or *1/*1 genotype , 1 had elevated lipase levels ( grade 3 ) and another had severe pain in the lower extremities ( grade 3 ) . CONCLUSION : These findings suggest that P22309 polymorphisms are important determinants of severe toxicity of nilotinib in Japanese patients . [ Vascular Calcification - Pathological Mechanism and Clinical Application - . The effect of cinacalcet on vascular calcification ] . DB01012 acts on calcium receptors ( CaR ) expressed on chief cells of the parathyroid gland to inhibit the secretion of parathyroid hormone ( PTH ) . This drug inhibits PTH secretion without causing an elevation of serum calcium and phosphorus , unlike active vitamin D . Several experimental studies demonstrated an inhibitory effect of calcimimetics on the progression of vascular calcification in animals with chronic kidney disease ( CKD ) , in keeping with the expression of the calcium-sensing receptor ( P41180 ) in vascular tissue . The EVOLVE , evaluated in patients with CKD 5D the effects of the cinacalcet on the progression of vascular calcification and hard cardiovascular outcomes , respectively . The EVOLVE trials missed their respective primary end point by intent-to-treat analysis . However , recently , in order to define the frequency of fatal and nonfatal cardiovascular events attributable to atherosclerotic and nonatherosclerotic mechanisms , risk factors for these events , and the effects of cinacalcet , post hoc analysis using adjudicated data collected during the EVOLVE Trial were perfomed . In this trial , combining fatal and nonfatal cardiovascular events , randomization to cinacalcet reduced the rates of sudden death and heart failure . Patients randomized to cinacalcet experienced fewer nonatherosclerotic cardiovascular events , while the effect of cinacalcet on atherosclerotic events did not reach statistical significance . Clinical utilization of cinacalcet in hypercalcemic conditions . INTRODUCTION : DB01012 has recently been introduced as a treatment for secondary hyperparathyroidism in dialysis patients and for parathyroid carcinoma . However , there has been an increasing interest in finding out whether cinacalcet can be used as a treatment for other parathyroid hormone ( PTH ) -dependent hypercalcemic conditions also . AREAS COVERED : The article reports the most relevant recent contributions dealing with calcium sensing receptor ( P41180 ) physiology as well as cinacalcet pharmacokinetics and pharmacodynamics . It also looks at the different hypercalcemic conditions where the use of cinacalcet has been proposed . This article was researched using clinical trials , case reports and outstanding basic research published in the last 3 years ( MEDLINE database up to 31 November 2010 ) . It provides the reader with an insight into the many unaddressed issues regarding cinacalcet that need to be resolved before it can be used in newly proposed fields . EXPERT OPINION : Since cinacalcet may not only have an effect on parathyroid P41180 but also on P41180 expressed at bone and renal levels , it can currently only be considered a good alternative to parathyroidectomy in PTH-dependent hypercalcemic conditions when surgical intervention is burdened by a high failure rate or when it can be considered a risky procedure . At present , cinacalcet can not be considered the first choice treatment in asymptomatic primary hyperparathyroidism or in mild-to-moderate forms of familial hypocalciuric hypocalcemia . The immunological effects of electrolyzed reduced water on the Echinostoma hortense infection in C57BL/6 mice . Electrolyzed reduced water ( ERW ) is widely used for drinking by people in Asia . The purpose of this study was to examine the immunological effect of ERW on the immunity of animals by supplying ERW to C57BL/6 mice infected with Echinostoma hortense metacercariae . In the non-infected groups , interleukin ( IL ) -4 ( p < 0.001 ) , P05113 , P22301 , IL-1beta , tumor necrosis factor ( P01375 ) -alpha and immunoglobulin ( Ig ) A expression of the group fed ERW ( ERW group ) increased in small intestine compared with the normal control group . In the case of infected groups , the group fed ERW ( ERW+E. hortense group ) showed the result that P05112 , P05113 , P22301 and Ig A expression increased , but IL-1beta and P01375 ( p < 0.001 ) decreased , and the number of goblet cells ( p < 0.001 ) and helix pomatia agglutinin ( Q9Y251 ) positive cells increased compared with the group without feeding ERW . However , adult worm recovery rate was markedly increased ( p < 0.05 ) . On the other hand , the expression of all the cytokines except P22301 in spleen was mildly increased but not significant statistically , and there was no significant difference in the numerical changes of white blood cell ( WBC ) . These results indicate that feeding ERW may have influence on the local immune response ( Th-1 type cytokines such as IL-1beta , P01375 ) in the small intestine but not on the systemic immune response . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . [ Medical treatment of primary hyperparathyroidism : role of calcimimetics ] . Primary hyperparathyroidism ( PHPT ) is a common endocrinological process , characterized by chronic elevation of serum concentrations of calcium and parathyroid hormone ( PTH ) . The only intervention able to cure the disease is parathyroidectomy . However , there are few valid medical alternatives for patients whose PHPT is unresolved by surgery , or in those with contraindications for surgery or who refuse the procedure . The discovery of calcium-sensing receptors ( CaSRs ) , which regulate PTH secretion according to extracellular calcium concentrations , has allowed specific anti-parathyroid drugs called calcimimetics to be designed . DB01012 is an allosteric modulator of P41180 that has demonstrated safety and efficacy in controlling serum calcium values and in reducing PTH levels in patients with PHPT . The exact role of calcimimetics in the overall management of PHPT is promising and should be considered in future clinical practice guidelines . Determinants of prognosis and response to therapy in colorectal cancer . Identification of the molecular determinants of 5-fluorouracil ( DB00544 ) and irinotecan ( CPT-11 ) efficacy and toxicity is critically important for the development of more efficient and less toxic treatment strategies for patients with colon cancer . We have identified molecular predictors of response to chemotherapy with DB00544 and survival in patients with advanced colorectal cancer . Low gene expression levels of thymidylate synthase ( TS ) , dihydropyrimidine dehydrogenase ( Q12882 ) , and thymidine phosphorylase ( TP ) are associated with response and survival . Preliminary data suggest that gene expression levels of topoisomerase I , P38936 , bcl-2 , and ICE may be predictive of response to therapy with CPT-11 . Increased toxicity seen in patients treated with CPT-11 may be explained by polymorphism in the P22309 gene , which is responsible for glucuronidation of the active metabolite of CPT-11 . A new member of the amphiphysin family connecting endocytosis and signal transduction pathways . Src homology 3 ( SH3 ) domains are conserved modules which participate in protein interaction by recognizing proline-rich motifs on target molecules . To identify new SH3-containing proteins , we performed a two-hybrid screen with a proline-rich region of human Q07889 . One of the specific Q07889 interacting clones that were isolated from a mouse brain cDNA library defines a new protein that was named amphiphysin 2 because of its homology to the previously reported amphiphysin . P49418 2 is expressed in a number of mouse tissues through multiple RNA transcripts . Here , we report the amino acid sequence of a brain form of amphiphysin 2 ( BRAMP2 ) encoded by a 2 . 5-kilobase mRNA . BRAMP2 associates in vitro with elements of the endocytosis machinery such as alpha-adaptin and dynamin . On a biosensor surface , the BRAMP2/dynamin interaction appeared to be direct and partly dependent on a proline-rich sequence of dynamin . Association with dynamin was also observed in PC12 cells after cell stimulation with nerve growth factor , suggesting that amphiphysin 2 may be connected to receptor-dependent signaling pathways . This hypothesis is strengthened by the ability of BRAMP2 to interact with the P38936 (ras) exchange factor SOS , in vitro , as a possible point of interconnection between the endocytic and signaling pathways . DB01012 upregulates calcium-sensing receptors of parathyroid glands in hemodialysis patients . BACKGROUND : DB01012 hydrochloride ( cinacalcet ) , a calcimimetic , has been shown to upregulate calcium-sensing receptor ( P41180 ) expression in parathyroid glands of rats with chronic renal insufficiency . However , the effect of cinacalcet on the reduced P41180 expression in human parathyroid glands remains to be elucidated . METHODS : Four normal parathyroid glands and 71 hyperplastic parathyroid glands from 18 hemodialysis patients with refractory secondary hyperparathyroidism ( SHPT ) treated with ( n = 10 ; cinacalcet group ) or without ( n = 8 ; conventional group ) cinacalcet were examined immunohistochemically with a specific antibody against P41180 . The expression level of P41180 was analyzed semiquantitatively . RESULTS : Compared with normal glands , the immunohistochemical expression of P41180 was decreased significantly in both the cinacalcet and conventional groups . In the cinacalcet group , the expression of P41180 was increased significantly compared with that in the conventional group ( 1.83 ± 0.14 vs. 0.87 ± 0.15 , p < 0.001 ) , even though the proportion of patients using vitamin D sterols and the mean administered dose of calcitriol equivalents were not significantly different between the two groups . The expression of P41180 was significantly decreased in the larger glands ( > 500 mg ) compared with that in the smaller glands ( < 500 mg ) in both groups ; furthermore , it was markedly decreased in areas of nodular hyperplasia compared with diffuse hyperplasia in the cinacalcet group . CONCLUSIONS : Our results indicate that cinacalcet upregulates the depressed expression of P41180 in hemodialysis patients with SHPT , and that insufficient expression of P41180 , especially in larger glands with advanced nodular hyperplasia , underlies the pathogenesis of SHPT in patients who are resistant to cinacalcet . P62158 regulates Ca2+-sensing receptor-mediated Ca2+ signaling and its cell surface expression . The Ca(2+)-sensing receptor ( P41180 ) is a member of family C of the GPCRs responsible for sensing extracellular Ca(2+) ( [Ca(2+)](o) ) levels , maintaining extracellular Ca(2+) homeostasis , and transducing Ca(2+) signaling from the extracellular milieu to the intracellular environment . In the present study , we have demonstrated a Ca(2+)-dependent , stoichiometric interaction between P62158 and a P62158 -binding domain ( CaMBD ) located within the C terminus of P41180 ( residues 871-898 ) . Our studies suggest a wrapping around 1-14-like mode of interaction that involves global conformational changes in both lobes of P62158 with concomitant formation of a helical structure in the CaMBD . More importantly , the Ca(2+)-dependent association between P62158 and the C terminus of P41180 is critical for maintaining proper responsiveness of intracellular Ca(2+) responses to changes in extracellular Ca(2+) and regulating cell surface expression of the receptor . DB01012 HCl prevents development of parathyroid gland hyperplasia and reverses established parathyroid gland hyperplasia in a rodent model of CKD . BACKGROUND : Secondary hyperparathyroidism ( sHPT ) represents an adaptive response to progressively impaired control of calcium , phosphorus and vitamin D in chronic kidney disease ( CKD ) . It is characterized by parathyroid hyperplasia and excessive synthesis and secretion of parathyroid hormone ( PTH ) . Parathyroid hyperplasia in uremic rats can be prevented by calcium-sensing receptor ( P41180 ) activation with the calcimimetic cinacalcet ( Sensipar®/Mimpara® ) ; however , it is unknown , how long the effects of cinacalcet persist after withdrawal of treatment or if cinacalcet is efficacious in uremic rats with established sHPT . METHODS : We sought to determine the effect of cinacalcet discontinuation in uremic rats and whether cinacalcet was capable of influencing parathyroid hyperplasia in animals with established sHPT . RESULTS : Discontinuation of cinacalcet resulted in reversal of the beneficial effects on serum PTH and parathyroid hyperplasia . In rats with established sHPT , cinacalcet decreased serum PTH and mediated regression of parathyroid hyperplasia . The cinacalcet-mediated decrease in parathyroid gland size was accompanied by increased expression of the cyclin-dependent kinase inhibitor P38936 . Prevention of cellular proliferation with cinacalcet occurred despite increased serum phosphorus and decreased serum calcium . CONCLUSIONS : The animal data provided suggest established parathyroid hyperplasia can be reversed by modulating P41180 activity with cinacalcet and that continued treatment may be necessary to maintain reductions in PTH . Calcimimetic and calcilytic drugs for treating bone and mineral-related disorders . The calcium-sensing receptor ( P41180 ) plays a pivotal role in regulating systemic Ca(2+) homeostasis and is a target for drugs designed to treat certain disorders of bone and mineral metabolism . Calcimimetics are agonists or positive allosteric modulators of the P41180 ; they inhibit parathyroid hormone ( PTH ) secretion and stimulate renal Ca(2+) excretion . The first calcimimetic drug is cinacalcet , a positive allosteric modulator of the P41180 that is approved for treating secondary hyperparathyroidism ( Q9HD23 ) in patients on renal replacement therapy and for some forms of primary Q9HD23 characterized by clinically significant hypercalcemia . DB01012 is also being investigated as a therapy for other hypercalcemic conditions and certain hypophosphatemic disorders . Calcilytics are P41180 inhibitors that stimulate the secretion of PTH and decrease renal excretion of Ca(2+) . Although calcilytics have failed thus far as anabolic therapies for osteoporosis , they are currently being evaluated as novel therapies for new indications involving hypocalcemia and/or hypercalciuria . Clinical utility of calcimimetics targeting the extracellular calcium-sensing receptor ( P41180 ) . Calcimimetics , which activate the extracellular calcium ( Ca(o)(2+) ) -sensing receptor in the parathyroid and other tissues participating in Ca(o)(2+) homeostasis , were the first described allosteric activators of a G-protein-coupled receptor . DB01012 , the only calcimimetic currently approved for human use , is used clinically for treating secondary hyperparathyroidism ( e.g. , overactivity of parathyroid glands ) in patients being dialyzed for chronic kidney disease . By sensitizing the parathyroids to Ca(o)(2+) , cinacalcet lowers the circulating parathyroid hormone ( PTH ) level . It also reduces serum calcium and phosphate , changes increasing the percentage of patients achieving the guidelines recommended by the National Kidney Foundation ( NKF ) for these minerals . Studies are underway addressing whether better adherence to these guidelines in patients receiving cinacalcet reduces cardiovascular disease and related mortality , which are both common is the dialysis population . The second approved use of cinacalcet is for treating hypercalcemia in patients with inoperable parathyroid carcinoma . In this setting , it provides the first medical therapy chronically lowering serum calcium concentration in this condition , albeit not to normal in most patients . Its effect on the long-term prognosis of these patients , if any , is presently unclear . " Off-label " administration of cinacalcet [ i.e. , not yet approved by the US Food and Drug Administration ( FDA ) ] effectively lowers serum calcium and/or PTH in various other forms of hyperparathyroidism and increases serum phosphate in renal phosphate-wasting syndromes by reducing PTH-induced phosphaturia . In the future , the drug could conceivably be utilized to modulate the activity of the P41180 in other tissues ( i.e. , kidney , colon ) in therapeutically desirable ways . DB01373 -sensing receptor gene polymorphism Arg990Gly and its possible effect on response to cinacalcet HCl . DB01012 , a novel calcimimetic compound , is effective in reducing parathyroid hormone ( PTH ) levels in approximately 70 % of patients with secondary hyperparathyroidism . However , interindividual variations in the dose required to achieve the treatment goal have been noted in clinical studies . Our investigation examined the genetic polymorphisms of the calcium-sensing receptor ( P41180 ) gene as one possible cause of the different responses to cinacalcet . We report data on seven end-stage renal failure patients who were treated with regular haemodialysis and who participated in clinical trials of cinacalcet . All patients had secondary hyperparathyroidism with baseline intact PTH ( iPTH ) levels greater than 600 pg/ml . Three patients were male and four female with a mean+/-SD age of 60+/-12 years . DNA was extracted from peripheral lymphocytes . An area in exon 7 of the P41180 gene was amplified by the polymerase chain reaction and sequenced . Mean+/-SD baseline iPTH was 1086+/-189 pg/ml . The five patients without Arg990Gly demonstrated a 29.7+/-4.0 % ( +/-SEM ) reduction in iPTH from individual baseline . One patient was found to be homozygous for the Arg990Gly polymorphism and another was heterozygous for both arginine and glycine alleles . The homozygous patient showed a significantly higher sensitivity to cinacalcet compared to the other patients ( P=0.003 ) with a 76.3+/-7.7 % reduction in iPTH from baseline . No polymorphisms were noted in codons 986 or 1011 . This preliminary study points to the possibility that patients homozygous for glycine at the 990 position in exon 7 of the P41180 may be more sensitive to the calcimimetic drug cinacalcet compared to those who are homozygous for arginine at that location . S-sulfhydration of Q02750 leads to P09874 activation and DNA damage repair . The repair of DNA damage is fundamental to normal cell development and replication . Hydrogen sulfide ( H2S ) is a novel gasotransmitter that has been reported to protect cellular aging . Here , we show that H2S attenuates DNA damage in human endothelial cells and fibroblasts by S-sulfhydrating Q02750 at cysteine 341 , which leads to P09874 activation . H2S-induced Q02750 S-sulfhydration facilitates the translocation of phosphorylated P27361 /2 into nucleus , where it activates P09874 through direct interaction . Mutation of Q02750 cysteine 341 inhibits P29323 phosphorylation and P09874 activation . In the presence of H2S , activated P09874 recruits P18887 and P49916 to DNA breaks to mediate DNA damage repair , and cells are protected from senescence . Epithelial calcium-sensing receptor activation by eosinophil granule protein analog stimulates collagen matrix contraction . BACKGROUND : Eosinophils reside in normal gastrointestinal tracts and increase during disease states . Receptors for eosinophil-derived granule proteins ( EDGPs ) have not been identified , but highly cationic molecules , similar to eosinophil proteins , bind extracellular calcium-sensing receptors ( CaSRs ) . We hypothesized that stimulation of CaSRs by eosinophil proteins activates epithelial cells . METHODS : Caco2 intestinal epithelial cells , AML14.3D10 eosinophils , wild-type ( WT ) human embryonic kidney 293 ( HEK293 ) cells not expressing CaSRs ( P29320 -WT ) , and P41180 -transfected HEK293 cells ( P29320 - P41180 ) were stimulated with an eosinophil protein analog poly-L-arginine ( PA ) and phosphorylated extracellular signal-regulated kinase (pERK)1 and pERK2 were measured . Functional activation was measured with collagen lattice contraction assays . RESULTS : Coculture of Caco2 cells with AML14.3D10 eosinophils augmented lattice contraction as compared with lattices containing Caco2 cells alone . PA stimulation of Caco2 lattices augmented contraction . P29320 - P41180 stimulation with PA or Ca(2+) resulted in greater pERK activation than that of stimulated P29320 -WT cells . PA stimulated greater P29320 - P41180 lattice contraction than unstimulated lattices . Contraction of PA-stimulated and PA-unstimulated P29320 -WT lattices did not differ . CONCLUSION : Exposure of intestinal epithelia to the EDGP analog PA stimulates P41180 -dependent P29323 phosphorylation and epithelial-mediated collagen lattice contraction . We speculate that EDGP release within the epithelial layers activates the P41180 receptor , leading to matrix contraction and tissue fibrosis . Cytological properties of stromal cells derived from giant cell tumor of bone ( GCTSC ) which can induce osteoclast formation of human blood monocytes without cell to cell contact . When human blood monocytes were cocultured with stromal cells derived from human giant cell tumor of bone ( GCTSC ) and a Millipore filter ( 0.4 microm ) was interposed between monocytes and GCTSC , multinucleated giant cell formation of monocytes was induced . The multinucleated giant cells have characters as osteoclast-like cells , indicating that a soluble osteoclast-inducing factor(s) is secreted from GCTSC expressing Q9Y6Q6 , O14788 / O14788 / O14788 and P78536 mRNA . Furthermore , O00300 / O00300 inhibited GCTSC-induced osteoclastogenesis , showing that the Q9Y6Q6 - O14788 system is involved in GCTSC-induced osteoclastogenesis and that soluble form of O14788 / O14788 induces osteoclasts from monocytes . GCTSC expressed the cytokine mRNAs such as P09603 , GM- P04141 , P08700 , P05112 , P05231 , and P01579 mRNAs . None of IL-1ralpha , IL-1alpha , IL-1beta , P60568 , P05112 , P22301 , Q14116 , P01375 , G- P04141 and P01579 could be detected in all culture media . A significant amount of P05231 could be detected in the culture media of all GCTSC . P10145 was found in the culture media of two GCTSC and two osteosarcoma-derived cells . P09603 was detected in all culture media . GCTSC express P41180 , and stimulation of GCTSC with either extracellular Ca(2+) or neomycin , agonist of P41180 , augmented the expression of O14788 . Some lines of GCTSC expressed alkaline phosphatase , osteocalcin and Cbfa1 , suggesting that GCTSC are intimately related to osteoblastic lineage . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain . P35354 induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways . The functional significance of protease-activated receptors ( PARs ) in endothelial cells is largely undefined , and the intracellular consequences of their activation are poorly understood . Here , we show that the serine protease thrombin , a P25116 -selective peptide ( TFLLRN ) , and SLIGKV ( P55085 -selective peptide ) induce cyclooxygenase-2 ( P35354 ) protein and mRNA expression in human endothelial cells without modifying P23219 expression . P35354 induction was accompanied by sustained production of 6-keto-PGF1alpha , the stable hydrolysis product of prostacyclin , and this was inhibited by indomethacin and the P35354 -selective inhibitor NS398 . P25116 and P55085 stimulation rapidly activated both P27361 /2 and p38MAPK , and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha formation . Thrombin and peptide agonists of P25116 and P55085 increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus , and this , as well as PAR-induced 6-keto-PGF1alpha synthesis , was inhibited by co-infection with adenovirus encoding wild-type or mutated ( Y42F ) P25963 . Thrombin- and SLIGKV-induced P35354 expression and 6-keto-PGF1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG-132 . Activation of P25116 or P55085 promoted nuclear translocation and phosphorylation of p65-NF-kappaB , and thrombin-induced but not P55085 -induced p65-NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK . Activation of Q96RI0 by AYPGKF increased phosphorylation of P27361 /2 and p38MAPK without modifying NF-kappaB activation or P35354 induction . Our data show that P25116 and P55085 , but not Q96RI0 , are coupled with P35354 expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on P27361 /2 , p38MAPK , and P25963 -dependent NF-kappaB activation . Dopamine modulating drugs influence striatal (+)-[11C]DTBZ binding in rats : Q05940 binding is sensitive to changes in vesicular dopamine concentration . Binding of (+)-[11C]DTBZ ( dihydrotetrabenazine ) to the striatal vesicular monoamine transporter ( Q05940 ) is widely considered to be a stable marker of dopamine neurone integrity . However , we now find that specific binding of a tracer dose of (+)-[11C]DTBZ is modestly increased in rat striatum following dopamine depletion with alpha-methyl-p-tyrosine ( alpha-MPT , +14 % ) or DB01576 ( d- P49418 , 20 mg/kg , +12 % ) and decreased following dopamine elevation with gamma-hydroxybutyrate ( DB01440 , -16 % ) or levodopa ( -20 % ) . We suggest that in vivo (+)-[11C]DTBZ binding in imaging studies is subject to competition by vesicular dopamine and , in this respect , is not a " stable " dopamine biomarker as is generally assumed . Calcimimetics in the chronic kidney disease-mineral and bone disorder . Mineral and bone disorders ( MBD ) are both an early and very common complication of chronic kidney disease ( CKD ) . It is now accepted that they represent a significant risk factor , explaining the high cardiovascular morbidity and mortality in CKD patients . During the last decade , we have been witnessing many advances in the nomenclature , classification , pathophysiology , diagnosis , and treatment of CKD and some of its complications , such as CKD-MBD . The identification of the calcium-sensing receptor ( P41180 ) involvement in the pathogenesis of primary and secondary hyperparathyroidism ( SHPT ) and the availability of a new class of drugs called calcimimetics are two outstanding examples . DB01012 , the only available calcimimetic , has been shown to be a very effective therapeutic tool in CKD-MBD . Many clinical trials with cinacalcet in hemodialysis patients with SHPT have shown a reduction in parathyroid hormone , calcium ( Ca ) , phosphate ( P ) and Ca x P product levels , allowing far greater success in reaching therapeutic goals as recommended by international guidelines . Additionally , some studies have shown that the use of cinacalcet may improve other aspects of CKD-MBD , reducing the risk of vascular calcification and parathyroidectomy , among others . Prospective studies on dialysis patients , with hard endpoint data , are currently underway . This review summarizes the most significant aspects of calcimimimetics based on both experimental and clinical results , underlining their possibilities not only for the treatment of isolated SHPT but also for other CKD-MBD related conditions . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . LPS induces cardiomyocyte injury through calcium-sensing receptor . DB01373 -sensing receptor ( P41180 ) belongs to the family C of G-protein coupled receptors . We have previously demonstrated that P41180 could induce apoptosis of cultured neonatal rat ventricular cardiomyocytes in simulated ischemia/reperfusion . It remains unknown whether the P41180 has function in lipopolysaccharide ( LPS ) -induced myocardial injure . The aim of this study was to investigate whether the P41180 plays a role in LPS-induced myocardial injury . Cultured neonatal rat cardiomyocytes were treated with LPS , with or without pretreatment with the P41180 -specific agonist gadolinium chloride ( GdCl3 ) or the P41180 -specific antagonist NPS2390 . Release of P01375 -α and P05231 from cardiomyocytes was observed . Levels of malonaldehyde ( MDA ) , lactate dehydrogenase ( LDH ) , and activity of superoxide dismutase ( SOD ) were measured . In addition , apoptosis of the cardiomyocytes , [Ca(2+)]i and level of P41180 expression were determined . The results showed that LPS increased cardiomyocytes apoptosis , [Ca(2+)]i , MDA , LDH , P01375 -α , P05231 release , and P41180 protein expression . Compared with LPS treatment alone , pretreatment with GdCl3 further increased apoptosis of cardiomyocytes , MDA , LDH , P01375 -α , P05231 release , [Ca(2+)]i , and the expression of the P41180 protein . Conversely , pretreatment with NPS2390 decreased apoptosis of cardiomyocytes , MDA , LDH , P01375 -α , P05231 release , [Ca(2+)]i and the expression of the P41180 protein . These results demonstrate that LPS could induce cardiomyocyte injury . Moreover , LPS-induced cardiomyocyte injury was related to P41180 -mediated cardiomyocytes apoptosis , P01375 -α , P05231 release , and increase of intracellular calcium . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Human SWI/SNF directs sequence-specific chromatin changes on promoter polynucleosomes . Studies in humans and other species have revealed that a surprisingly large fraction of nucleosomes adopt specific positions on promoters , and that these positions appear to be determined by nucleosome positioning DNA sequences ( NPSs ) . Recent studies by our lab , using minicircles containing only one nucleosome , indicated that the human SWI/SNF complex ( hSWI/SNF ) prefers to relocate nucleosomes away from NPSs . We now make use of novel mapping techniques to examine the hSWI/SNF sequence preference for nucleosome movement in the context of polynucleosomal chromatin , where adjacent nucleosomes can limit movement and where hSWI/SNF forms altered dinucleosomal structures . Using two P0C0P6 templates ( 5S rDNA and 601 ) and two hSWI/SNF target promoter templates ( c-myc and P22309 ) , we observed hSWI/SNF-driven depletion of normal mononucleosomes from almost all positions that were strongly favored by assembly . In some cases , these mononucleosomes were moved to hSWI/SNF-preferred sequences . In the majority of other cases , one repositioned mononucleosome appeared to combine with an unmoved mononucleosome forming a specifically localized altered or normal dinucleosome . These effects result in dramatic , template-specific changes in nucleosomal distribution . Taken together , these studies indicate hSWI/SNF is likely to activate or repress transcription of its target genes by generating promoter sequence-specific changes in chromatin configuration . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . DB01012 Treatment in an Adolescent With Concurrent 22q11.2 Deletion Syndrome and Familial Hypocalciuric Hypercalcemia Type 3 Caused by P53680 Mutation . CONTEXT : The 22q11.2 deletion syndrome ( DS ) is a common multiple anomaly syndrome in which typical features include congenital heart defects , facial dysmorphism , and palatal anomalies . Hypocalcemia due to hypoparathyroidism is a common endocrine manifestation resulting from variable parathyroid hypoplasia , but hypercalcemia has not previously been reported in 22q11.2 DS . CASE DESCRIPTION : Our patient is a 16-year-old adolescent male with dysmorphic facial features and delayed motor and speech development . At 2 years of age , 22q11.2 DS was confirmed by fluorescence in situ hybridization . In contrast to hypoparathyroidism that is usually seen in 22q11.2 DS , this patient had early childhood-onset hypercalcemia with inappropriately high PTH levels and hypocalciuria . Genomic DNA was obtained from the proband and screened for calcium-sensing receptor ( P41180 ) mutations with negative results . No parathyroid tissue could be localized by imaging or surgical exploration . As a result of symptomatic hypercalcemia , the patient was treated with a calcimimetic ( cinacalcet ) . During the treatment , plasma calcium normalized with mild symptoms of hypocalcemia . After discontinuation of cinacalcet , calcium returned to high pretreatment levels . Further DNA analysis of adaptor protein-2 σ subunit ( P53680 ) showed a heterozygous missense mutation c.44 G > T , resulting in a p.R15L substitution ; the mutation was absent in the healthy parents and two siblings . CONCLUSIONS : Hypercalcemia in our patient with 22q11.2 DS could be explained by the de novo mutation in P53680 . Identification of a genetic cause for hypercalcemia is helpful in guiding management and avoiding unnecessary treatment . Antagonizing amyloid-β/calcium-sensing receptor signaling in human astrocytes and neurons : a key to halt Alzheimer 's disease progression ? Astrocytes ' roles in late-onset Alzheimer 's disease ( LOAD ) promotion are important , since they survive soluble or fibrillar amyloid-β peptides ( Aβs ) neurotoxic effects , undergo alterations of intracellular and intercellular Ca(2+) signaling and gliotransmitters release via the Aβ/α7-nAChR ( α7-nicotinic acetylcholine receptor ) signaling , and overproduce/oversecrete newly synthesized Aβ42 oligomers , NO , and P15692 via the Aβ/ P41180 ( calcium-sensing receptor ) signaling . Recently , it was suggested that the NMDAR ( N-methyl-D-aspartate receptor ) inhibitor nitromemantine would block the synapse-destroying effects of Aβ/α7-nAChR signaling . Yet , this and the progressive extracellular accrual and spreading of Aβ42 oligomers would be stopped well upstream by P0C0P6 2143 , an allosteric P41180 antagonist ( calcilytic ) . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Synaptic vesicular monoamine transporter expression : distribution and pharmacologic profile . The human vesicular monoamine transporter ( hSVMT ) cDNA predicts a protein of 515 amino acids that shares 92 % amino acid identity with the rat cDNA . Northern analyses reveal expression of 4.3 kb Q05940 mRNAs in rat hypothalamus , midbrain and brainstem , a 3 kb hSVMT mRNA in human brainstem and a 4.8 kb hSVMT mRNA in human hypothalamus . In situ hybridization documents significant Q05940 expression in human nigra compacta neurons and in rat hypothalamic neurons whose distribution patterns are identical to those previously reported to display histaminergic markers . COS cell hSVMT expression yielded nanomolar affinities for tetrabenazine and reserpine , micromolar affinities for haloperidol , GBR12909 , serotonin , mazindol , nomifensin and DB01576 , while dopamine , epinephrine , norepinephrine and histamine each displayed millimolar affinities . These observations extend the pharmacological characterization of hSVMT and studies of its distribution , and indicate likely physiological roles for Q05940 in packaging monoamine transmitters including histamine . DB01012 : pharmacological and clinical aspects . The calcium sensing receptor ( P41180 ) is expressed in cells secreting calcium-regulating hormones , in cells involved in calcium transport and in many other tissues , with an as yet not completely defined role . In parathyroid cells , the P41180 stimulation inhibits parathyroid hormone ( PTH ) secretion , synthesis and parathyroid cell proliferation . DB01012 belongs to calcimimetic type II compounds that can interact with P41180 , increasing its affinity for calcium . Clinical studies have proved cinacalcet to be effective in reducing calcium and PTH levels in primary hyperparathyroidism and in reducing PTH , calcium and phosphate in patients with secondary hyperparathyroidism owing to chronic renal failure , with a relatively safe profile , the only reported adverse events being hypocalcaemia and gastrointestinal symptoms . However , though calcimimetics do represent a real advancement in the field of the treatment of PTH secretion disturbances , there is a need for clinical trials , which should aim to demonstrate that a better control of biochemical parameters is also matched with better clinical outcomes . Translational research in bipolar disorder : emerging insights from genetically based models . Bipolar disorder ( BPD ) is characterized by vulnerability to episodic depression and mania and spontaneous cycling . Because of marked advances in candidate-gene and genome-wide association studies , the list of risk genes for BPD is growing rapidly , creating an unprecedented opportunity to understand the pathophysiology of BPD and to develop novel therapeutics for its treatment . However , genetic findings are associated with major unresolved issues , including whether and how risk variance leads to behavioral abnormalities . Although animal studies are key to resolving these issues , consensus is needed regarding how to define and monitor phenotypes related to mania , depression and mood swing vulnerability in genetically manipulated rodents . In this study we discuss multiple facets of this challenging area , including theoretical considerations , available tests , limitations associated with rodent behavioral modeling and promising molecular-behavioral findings . These include O15516 , glycogen synthase kinase 3beta ( GSK-3beta ) , glutamate receptor 6 ( Q13002 ) , extracellular signal-regulated kinase-1 ( P27361 ) , p11 ( or P60903 ) , vesicular monoamine transporter 2 ( Q05940 or Q05940 ) , glucocorticoid receptors ( GRs ) , Bcl-2-associated athanogene-1 ( Q99933 ) and mitochondrial DNA polymerase-gamma ( P54098 ) . Some mutant rodent strains show behavioral clusters or activity patterns that cross-species phenocopy objective/observable facets of mood syndromes , and changes in these clustered behaviors can be used as outcome measures in genetic-behavioral research in BPD . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . Inactivation of CaMIT1 inhibits Candida albicans phospholipomannan beta-mannosylation , reduces virulence , and alters cell wall protein beta-mannosylation . Studies on Candida albicans phospholipomannan have suggested a novel biosynthetic pathway for yeast glycosphingolipids . This pathway is thought to diverge from the usual pathway at the mannose-inositol-phospho-ceramide ( MIPC ) step . To confirm this hypothesis , a C. albicans gene homologue for the Saccharomyces cerevisiae Q09428 gene was identified and named MIT1 as it coded for GDP-mannose:inositol-phospho-ceramide mannose transferase . Two copies of this gene were disrupted . Western blots of cell extracts revealed that strain mit1Delta contained no PLM . Thin layer chromatography and mass spectrometry confirmed that mit1Delta did not synthesize MIPC , demonstrating a role of MIT1 in the mannosylation of C. albicans IPCs . As MIT1 disruption prevented downstream beta-1,2 mannosylation , mit1Delta represents a new C. albicans mutant affected in the expression of these specific virulence attributes , which act as adhesins/immunomodulators . mit1Delta was less virulent during both the acute and chronic phases of systemic infection in mice ( 75 and 50 % reduction in mortality , respectively ) . In vitro , mit1Delta was not able to escape macrophage lysis through down-regulation of the P27361 /2 phosphorylation pathway previously shown to be triggered by PLM . Phenotypic analysis also revealed pleiotropic effects of MIT1 disruption . The most striking observation was a reduced beta-mannosylation of phosphopeptidomannan . Increased beta-mannosylation of mannoproteins was observed under growth conditions that prevented the association of beta-oligomannosides with phosphopeptidomannan , but not with PLM . This suggests that C. albicans has strong regulatory mechanisms associating beta-oligomannoses with different cell wall carrier molecules . These mechanisms and the impact of the different presentations of beta-oligomannoses on the host response need to be defined . Impact of clinically relevant mutations on the pharmacoregulation and signaling bias of the calcium-sensing receptor by positive and negative allosteric modulators . DB01012 is predominantly used to treat secondary hyperparathyroidism due to end-stage renal failure , but , more recently , its potential clinical efficacy in treating patients with loss-of-function mutations in the calcium-sensing receptor ( P41180 ) has been recognized . Many clinically relevant P41180 mutations are located in the heptahelical membrane spanning and extracellular loop regions of the receptor , where allosteric modulators are predicted to bind . The aim of the present study was to investigate the impact of such mutations on the pharmacoregulation of the P41180 by the positive and negative allosteric modulators , cinacalcet and P0C0P6 -2143 , respectively . Both cinacalcet and P0C0P6 -2143 effectively rescued mutants whose cell surface expression was substantially impaired , suggesting that both classes of drug can stabilize a receptor conformation that is trafficked more effectively to the cell surface . In addition , functional impairments in almost all mutant CaSRs were rescued by either cinacalcet or P0C0P6 -2143 via restoration of intracellular signaling . There was a significantly greater ability of both compounds to modulate agonist-stimulated intracellular Ca(2+) mobilization than P27361 /2 phosphorylation , indicating that the allosteric modulators engender bias in agonist-stimulated P41180 signaling to different pathways . Three mutations ( G(670)R , P(748)R , and L(773)R ) altered the binding affinity of allosteric modulators to the P41180 , and 3 mutations ( V(817)I , L(773)R , and E(767)K ) altered the cooperativity between the allosteric modulator and Ca(2+)(o) . These findings have important implications for the treatment of diseases associated with P41180 mutations using allosteric P41180 modulators and for analyzing the effects of mutations on the function and pharmacoregulation of the P41180 . Abnormal response to stress and impaired P0C0P6 -induced hyperlocomotion , anxiolytic effect and corticosterone increase in mice lacking Q6W5P4 . Q6W5P4 is a G protein coupled receptor expressed in multiple brain regions involved in modulation of stress . Central administration of P0C0P6 , the putative endogenous ligand of Q6W5P4 , can induce hyperlocomotion , anxiolytic effects and activation of the Q9Y251 axis . The role of Q6W5P4 in the brain remains unsettled . Here we used Q6W5P4 gene-targeted mice to define the functional role of Q6W5P4 under basal conditions on locomotion , anxiety- and/or depression-like behavior , corticosterone levels , acoustic startle with prepulse inhibition , learning and memory , and under P0C0P6 -induced locomotor activation , anxiolysis , and corticosterone release . Male , but not female , Q6W5P4 -deficient mice exhibited enhanced depression-like behavior in a forced swim test , reduced acoustic startle response , and minor changes in the Morris water maze . Neither male nor female Q6W5P4 -deficient mice showed alterations of baseline locomotion , anxiety-like behavior , or corticosterone release after exposure to a forced swim test or methamphetamine challenge in an open-field . After intracerebroventricular ( ICV ) administration of P0C0P6 , Q6W5P4 -deficient mice failed to show normal P0C0P6 -induced increases in locomotion , anxiolysis , or corticosterone release compared with WT P0C0P6 -treated mice . These findings demonstrate that Q6W5P4 is essential in mediating P0C0P6 effects on behavior . Induction of G2/M arrest and inhibition of cyclooxygenase-2 activity by curcumin in human bladder cancer T24 cells . Curcumin , a polyphenol compound derived from Curcuma longa Linn , has been recognized as a promising anti-cancer drug due to its multiple properties including anti-inflammatory , anti-oxidant and anti-carcinogenic activities . To elucidate the mechanisms by which curcumin inhibits human bladder carcinoma T24 cell proliferation , we tested the effects of curcumin on specific cell cycle pathways and on the expression of cyclooxygenases ( COXs ) . Curcumin inhibited the growth of T24 cells and induced G2/M arrest in a concentration-dependent manner , effects associated with the down-regulation of cyclin A and up-regulation of cyclin-dependent kinase ( Cdk ) inhibitor P38936 ( P38936 /CIP1 ) . However , other G2/M regulatory molecules , such as cyclin A , Cdc2 , Cdk2 , Wee1 and Cdc25C , were not modulated by curcumin treatment . Furthermore , curcumin decreased the levels of P35354 mRNA and protein expression without significant changes in the levels of P23219 , which correlated with a decrease in prostaglandin E2 ( DB00917 ) synthesis . These observations suggest that curcumin may have therapeutic potential for bladder cancer patients . Xaliproden ( SR57746A ) induces P08908 receptor-mediated Q96HU1 kinase activation in PC12 cells . Neurotrophic growth factors are involved in cell survival . However , natural growth factors have a very limited therapeutic use because of their short half-life . In the present study , we investigated the mechanism of action of a non-peptidic neurotrophic drug , Xaliproden , a potential molecule for the treatment of motoneuron diseases , since the transduction pathways of this synthetic P08908 agonist are very poorly understood . Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the P27361 and P28482 isoforms of Q96HU1 kinase , which then rapidly decrease to the basal level . We demonstrate that isoforms of the P29353 adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced Q96HU1 kinases activation . The inhibitor of Ras farnesylation , FPT-1 , and the protein kinase C inhibitors , GF 109203X and chelerythrine , inhibited the Xaliproden-induced Q96HU1 kinase activation , suggesting p21Ras and PKC involvement . Moreover , the observations that the P08908 antagonist , pindobind , and pertussis toxin abolished the Xaliproden-induced P29323 stimulation suggested that Xaliproden activates the Q96HU1 kinase pathways by stimulating the G protein-coupled receptor , P08908 . These results demonstrate clearly that the non-peptidic compound , Xaliproden , exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins . These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by Q96HU1 kinase pathway by a pertussis toxin-sensitive mechanism involving P08908 receptors , P38936 Ras and MEK-1 and by PKC and Akt pathways . Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins . Poly(ADP-ribose) is formed in possibly all multicellular organisms by a familiy of poly(ADP-ribose) polymerases ( PARPs ) . P09874 , the best understood and until recently the only known member of this family , is a DNA damage signal protein catalyzing its automodification with multiple , variably sized ADP-ribose polymers that may contain up to 200 residues and several branching points . Through these polymers , P09874 can interact noncovalently with other proteins and alter their functions . Here we report the discovery of a poly(ADP-ribose)-binding sequence motif in several important DNA damage checkpoint proteins . The 20-amino acid motif contains two conserved regions : ( i ) a cluster rich in basic amino acids and ( ii ) a pattern of hydrophobic amino acids interspersed with basic residues . Using a combination of alanine scanning , polymer blot analysis , and photoaffinity labeling , we have identified poly(ADP-ribose)-binding sites in the following proteins : p53 , P38936 (CIP1/ P38936 ) , xeroderma pigmentosum group A complementing protein , P52701 , P49916 , P18887 , DNA polymerase epsilon , DNA-PK(CS) , P12956 , NF-kappaB , inducible nitric-oxide synthase , caspase-activated DNase , and telomerase . The poly(ADP-ribose)-binding motif was found to overlap with five important functional domains responsible for ( i ) protein-protein interactions , ( ii ) DNA binding , ( iii ) nuclear localization , ( iv ) nuclear export , and ( v ) protein degradation . Thus , PARPs may target specific signal network proteins via poly(ADP-ribose) and regulate their domain functions . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Novel strategies in drug discovery of the calcium-sensing receptor based on biased signaling . A hallmark of chronic kidney disease is hyperphosphatemia due to renal phosphate retention . Prolonged parathyroid gland exposure to hyperphosphatemia leads to secondary hyperparathyroidism characterized by hyperplasia of the glands and excessive secretion of parathyroid hormone ( PTH ) , which causes renal osteodystrophy . PTH secretion from the parathyroid glands is controlled by the calcium-sensing receptor ( P41180 ) that senses extracellular calcium . High extracellular calcium activates the P41180 causing inhibition of PTH secretion through multiple signaling pathways . DB01012 is the first drug targeting the P41180 and can be used to effectively control and reduce PTH secretion in PTH-related diseases . DB01012 is a positive allosteric modulator of the P41180 and affects PTH secretion from parathyroid glands by shifting the calcium-PTH concentration-response curve to the left . One major disadvantage of cinacalcet is its hypocalcemic side effect , which may be caused by increased P41180 -mediated calcitonin secretion from the thyroid gland . However , multiple studies indicate that PTH and calcitonin secretion are stimulated by different signaling pathways , and therefore it might be possible to develop a P41180 activating drug that selectively activates signaling pathways that inhibit PTH secretion while having no effect on signaling pathways involved in calcitonin secretion . Such a drug would have the same therapeutic value as cinacalcet in lowering PTH secretion while eliminating the side effect of hypocalcemia by virtue of it not affecting calcitonin secretion . The present review will focus on recent advancements in understanding signaling and biased signaling of the P41180 , and how that may be utilized to discover new and smarter drugs targeting the P41180 . γ-Glutamyl cysteine and γ-glutamyl valine inhibit P01375 -α signaling in intestinal epithelial cells and reduce inflammation in a mouse model of colitis via allosteric activation of the calcium-sensing receptor . BACKGROUND : The extracellular calcium-sensing receptor ( P41180 ) is distributed throughout the gastrointestinal tract , and its activation has been shown to promote intestinal homeostasis , suggesting that P41180 may be a promising target for novel therapies to prevent chronic intestinal inflammation such as inflammatory bowel disease ( Q9UKU7 ) . The γ-glutamyl dipeptides γ-glutamyl cysteine ( γ-EC ) and γ-glutamyl valine ( γ-EV ) are dietary flavor enhancing compounds , and have been shown to activate P41180 via allosteric ligand binding . The aim of this study was to examine the anti-inflammatory effects of γ-EC and γ-EV in vitro in intestinal epithelial cells and in a mouse model of intestinal inflammation . RESULTS : In vitro , treatment of Caco-2 cells with γ-EC and γ-EV resulted in the P41180 -mediated reduction of P01375 -α-stimulated pro-inflammatory cytokines and chemokines including P10145 , P05231 , and IL-1β , and inhibited phosphorylation of JNK and IκBα , while increasing expression of P22301 . In vivo , using a mouse model of dextran sodium sulfate ( DSS ) -induced colitis , γ-EC and γ-EV treatment ameliorated DSS-induced clinical signs , weight loss , colon shortening and histological damage . Moreover , γ-EC and γ-EV reduced the expression of P01375 -α , P05231 , P27352 -γ , IL-1β , and Q16552 , and increased the expression of P22301 in the colon , in a P41180 -dependent manner . The P41180 -mediated anti-inflammatory effects of γ-EC were abrogated in β-arrestin2 knock-down Caco-2 cells , and involvement of β-arrestin2 was found to inhibit P01375 -α-dependent signaling via cross-talk with the P01375 -α receptor ( TNFR ) . CONCLUSIONS : Thus P41180 activation by γ-EC and γ-EV can aid in maintaining intestinal homeostasis and reducing inflammation in chronic inflammatory conditions such as Q9UKU7 . Novel target for induction of apoptosis by cyclo-oxygenase-2 inhibitor SC-236 through a protein kinase C-beta(1)-dependent pathway . Nonsteroidal anti-inflammatory drugs ( NSAIDs ) reduce the risk of gastrointestinal cancers . Recently , a similar protective effect has been demonstrated by the specific cyclo-oxygenase-2 ( P35354 ) inhibitors . However , the exact mechanism that accounts for the anti-proliferative effect of specific P35354 inhibitors is still not fully understood , and it is still controversial whether these protective effects are predominantly mediated through the inhibition of P35354 activity and prostaglandin synthesis . Identification of molecular targets regulated by P35354 inhibitors could lead to a better understanding of their pro-apoptotic and anti-neoplastic activities . In the present study , we investigated the effect and the possible molecular target of a P35354 -specific inhibitor SC-236 on gastric cancer . We showed that SC-236 induced apoptosis in gastric cancer cells . However , this effect was not dependent on P35354 inhibition . SC-236 down-regulated the protein expression and kinase activity of P05771 (1) , increased the expression of PKCdelta and PKCeta , but did not alter the expression of other PKC isoforms in AGS cells . Moreover , exogenous prostaglandins or PGE(2) receptor antagonists could not reverse the inhibition effect on PKCbeta(1) by SC-236 , which suggested that this effect occurred through a mechanism independent of cyclo-oxygenase activity and prostaglandin synthesis . Overexpression of PKCbeta(1) attenuated the apoptotic response of AGS cells to SC-236 and was associated with overexpression of P38936 (waf1/cip1) . Inhibition of PKCbeta(1)-mediated overexpression of P38936 (waf1/cip1) partially reduced the anti-apoptotic effect of PKCbeta(1) . The down-regulation of PKCbeta(1) provides an explanation for P36551 -independent apoptotic effects of specific P35354 inhibitor in cultured gastric cancer cells . We also suggest that PKCbeta(1) act as survival mediator in gastric cancer , and its down-regulation by P35354 inhibitor SC-236 may provide new target for future treatment of gastric cancer . P00734 in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 ) has not been investigated . We determined the concentration of prothrombin in P04141 with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit -- 0.7 ng/ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 was 0.55 mg/l ( CV +/- 33 % , range : 0.28-0.93 mg/l ) , in normal plasma 121.8 mg/l +/- 21 % , resulting in a mean P04141 /plasma concentration quotient ( Q(Proth) -- 4.5 x 10(-3) ( CV +/- 35 % , range : 2.1-8.3 x 10(-3) ) corresponding to a mean albumin quotient in this group of subjects of Q(Alb) = 5.8 x 10(-3) . Due to the Q(Proth) and the molecular weight of prothrombin ( 72 kDa ) -- similar to that of albumin -- we conclude that prothrombin in normal human P04141 originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 if the blood- P04141 barrier function is normal .

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Combination effects of paclitaxel with signaling inhibitors in endometrial cancer cells . This study was conducted to evaluate and compare molecular and cellular effects of paclitaxel in combination with epidermoid growth factor receptor ( P00533 ) or/and mammalian target of rapamycin ( P42345 ) inhibitors with two endometrial cancer lines O14777 -1A and Ishikawa . Treatment was with the P00533 inhibitor RG14620 , the P42345 inhibitor rapamycin , and the conventional cytotoxic drug paclitaxel , alone or in combination . The 50 % inhibitory concentration ( IC50 ) and cell viability were determined by the MTT assay . Multiple drug effect/ combination indexes ( CI ) analysis was applied to assess interactions between paclitaxel and the two inhibitors . Apoptosis and cell cycling were evaluated by flow cytometry analysis . Western blotting was performed to evaluate the related protein alteration in PI3K/AKT signaling pathway . RG14620 , rapamycin and paclitaxel showed obvious dose-dependent growth inhibition with time . The IC50 of paclitaxel at 24 hours decreased significantly when pretreated with low doses of RG14620 and DB00877 alone or in combination . Moreover , combination index ( CI ) of paclitaxel with each inhibitor was larger than 1 , indicating a synergistic effect between pairs of drugs in these two cell lines . FACS analysis showed the cell apoptosis rate increased with a synergistic effect . On Western blotting , activation of PI3K/AKT pathway was detected in both two cell lines in the control case . When paclitaxel was used as a single-agent or in combinations , the protein expression of PI3K/AKT pathway totally abated , especially in O14777 -1A cells , suggesting a role in chemoresistance . The combination of three drugs induced the greatest over-expression of caspase-3 . Combining targeted inhibitors with cytotoxic chemotherapy appears to be a promising strategy for the effective treatment of endometrial cancer which merits further clinical investigation . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . Generic bioaffinity silicone surfaces . Synthetic polymer surfaces require surface modification to improve biocompatibility . A generic route to biocompatible silicone elastomers is described involving high yield surface functionalization of standard silicones with hydrosilanes , hydrosilylation using asymmetric , allyl- , NSC-terminated DB09287 of narrow molecular weight , and covalent modification in one step with amine-containing biological molecules including oligopeptides ( YIGSR , RGDS ) , proteins ( P01133 , albumin , fibrinogen , mucin ) , and glycosaminoglycans ( heparin ) . Efficient , high-density binding ( e.g. , 0.2 P01133 molecules/nm2 ) was demonstrated using radiolabeling studies . The resulting surfaces were demonstrated to be biocompatible by further reaction with biomolecules , for example , thrombosis suppression on surfaces modified by heparin + P01008 , and the formation of confluent corneal epithelial cell layers on P01133 , RGDS , or YIGSR surfaces . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . Boca-dependent maturation of beta-propeller/ P01133 modules in low-density lipoprotein receptor proteins . The extracellular portions of cell surface receptor proteins are often comprised of independently folding protein domains . As they are translated into the endoplasmic reticulum ( ER ) , some of these domains require protein chaperones to assist in their folding . Members of the low-density lipoprotein receptor ( P01130 ) family require the chaperone called Boca in Drosophila or its ortholog , Mesoderm development , in the mouse . All LDLRs have at least one six-bladed beta-propeller domain , which is immediately followed by an epidermal growth factor ( P01133 ) repeat . We show here that Boca is specifically required for the maturation of these beta-propeller/ P01133 modules through the secretory pathway , but is not required for other P01130 domains . Protein interaction data suggest that as LDLRs are translated into the ER , Boca binds to the beta-propeller . Subsequently , once the P01133 repeat is translated , the beta-propeller/ P01133 module achieves a more mature state that has lower affinity for Boca . We also show that Boca-dependent beta-propeller/ P01133 modules are found not only throughout the P01130 family but also in the precursor to the mammalian P01133 ligand . Combined Effects of Q07869 γ Agonists and Epidermal Growth Factor Receptor Inhibitors in Human Proximal Tubule Cells . We aimed to determine whether epidermal growth factor receptor ( P00533 ) inhibition , in addition to a peroxisome proliferator-activated receptor gamma ( Q07869 γ ) agonist , prevents high-glucose-induced proximal tubular fibrosis , inflammation , and sodium and water retention in human proximal tubule cells exposed to normal glucose ; high glucose ; high glucose with the Q07869 γ agonist pioglitazone or with the P- P00533 inhibitor , gefitinib ; or high glucose with both pioglitazone and gefitinib . We have shown that high glucose increases AP-1 and NF κ B binding activity , downstream phosphorylation of P00533 and Erk1/2 , and fibronectin and collagen IV expression . Pioglitazone reversed these effects but upregulated P48764 and P29972 expression . Gefitinib inhibited high glucose induced fibronectin and collagen IV , and P00533 and Erk1/2 phosphorylation and reversed pioglitazone-induced increases in P48764 and P29972 expression . Our data suggests that combination of an P00533 inhibitor and a Q07869 γ agonist mitigates high-glucose-induced fibrosis and inflammation and reverses the upregulation of transporters and channels involved in sodium and water retention in human proximal tubule cells . Hence P00533 blockade may hold promise , not only in limiting tubulointerstitial pathology in diabetic nephropathy , but also in limiting the sodium and water retention observed in patients with diabetes and exacerbated by Q07869 γ agonists . DB00281 inhibits tyrosine kinase activity of the epidermal growth factor receptor and suppresses proliferation of corneal epithelial cells . BACKGROUND : Although lidocaine is recognized as an excellent topical corneal analgesic , its toxic effect on corneal epithelial cells limits its use during corneal epithelial wound healing . Mechanism of the impairment of corneal reepithelialization with lidocaine , however , has not been evaluated . The authors ' previous study revealed that lidocaine inhibits the activity of tyrosine kinase receptors through the interaction with specific amino acid sequences around autophosphorylation sites , including acidic , basic , and aromatic amino acids . P00533 ( P00533 ) , a tyrosine kinase receptor with an important role in epithelial cell proliferation after corneal wounding , also possesses these amino acids sequences around autophosphorylation sites . The authors hypothesized that lidocaine would suppress tyrosine kinase activity of P00533 and would impair corneal epithelial cell proliferation . METHODS : To investigate the effect of lidocaine ( 4 microM-40 mM ) on epidermal growth factor ( P01133 ) -stimulated autophosphorylation of P00533 , the authors studied purified P00533 in microtubes . They cultured human corneal epithelial cells ( HCECs ) with P01133 and lidocaine to investigate the effect of lidocaine on cell proliferation and on autophosphorylation of P00533 in HCECs . RESULTS : DB00281 ( > or =400 microM ) significantly suppressed P01133 -stimulated autophosphorylation of the purified P00533 . In the HCEC study , P01133 alone stimulated cell proliferation and increased autophosphorylation of P00533 in HCECs . DB00281 ( > or = 400 microM ) significantly suppressed both the proliferation of HCECs promoted by P01133 and P01133 -stimulated autophosphorylation of P00533 . CONCLUSION : DB00281 directly inhibits tyrosine kinase activity of P00533 and suppresses the corneal epithelial cell proliferation . The effectiveness of lurasidone as an adjunct to lithium or divalproex in the treatment of bipolar disorder . The majority of patients with bipolar disorder spend a lot of time in depressive episodes that impose a great burden on patients , caregivers , and society and accounts for the largest part of the morbidity-mortality of the illness . DB08815 is an atypical antipsychotic with a potent binding affinity as antagonist for D2 , 5- Q13049 , P34969 , and partial agonist at P08908 receptors . Affinity for other receptors as H1 and muscarinic were negligible . DB08815 was approved in 2010 for the treatment of schizophrenia and recently , 2013 , for bipolar depression in monotherapy and an adjunct to lithium or valproate . Clinical trials have established that lurasidone adjuvant to lithium or valproate has more efficacy than the placebo and is associated with minimal weight gain and no clinically meaningful alterations in glucose , lipids , or the QT interval . Additional studies are desirable to know the clinical profile of lurasidone in long-term treatment , in patients with bipolar II disorders , and versus other antipsychotic agents . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . The antiproliferative effect of lidocaine on human tongue cancer cells with inhibition of the activity of epidermal growth factor receptor . Local anesthetics suppress proliferation in several cancer cells . The mechanism of the suppression , however , is unknown . Our previous study shows that lidocaine , at the level of tissue concentration under topical or local administration , has a direct inhibitory effect on the activity of epidermal growth factor receptor ( P00533 ) , which is a potential target for antiproliferation in cancer cells . Therefore , we hypothesized that lidocaine would suppress the proliferation of cancer cells through the inhibition of P00533 activity . We investigated the effects of lidocaine ( 40-4000 microM ) on proliferation of a human tongue cancer cell line , CAL27 , which has a high level of P00533 expression , and also examined the effect of lidocaine on epidermal growth factor ( P01133 ) -stimulated autophosphorylation of P00533 in CAL27 cells . A clinical concentration of lidocaine ( 400 microM ) suppressed both serum-induced and P01133 -induced proliferation of CAL27 cells and inhibited P01133 -stimulated tyrosine kinase activity of P00533 without cytotoxicity . A larger concentration of lidocaine ( 4000 microM ) showed cytotoxicity with an antiproliferative effect . We suggest that the inhibition of P01133 -stimulated P00533 activity is one of the mechanisms of the antiproliferative effect of lidocaine on CAL27 cells . DB00281 administered topically within the oral cavity for cancer pain relief may suppress the proliferation of human tongue cancer cells . Specific properties of a C-terminal truncated androgen receptor detected in hormone refractory prostate cancer . Mutations in the human androgen receptor ( AR ) gene that lead to C-terminus truncated AR variants are frequently detected in prostate cancer ( PC ) . These AR variants lack both the ligand-binding domain ( LBD ) and the AF-2 region . The aim of this study was to delineate the alternative mechanisms that lead to the activation of such AR variants as they are unresponsive to hormone stimulation , and to outline consequences of the loss of the LBD/AF-2 region on their functional properties . By using an MMTV-luciferase reporter construct and LY294002 , UO126 , or ZD1839 , inhibitor of PI3K , Q02750 /2 , and P00533 signaling pathway respectively , we demonstrated that phosphorylation was required for full transcriptional activities of one these AR variants , the Q640X mutant AR . Western-blot analyses confirmed that these inhibitors affect the phosphorylation status of this AR variant . Furthermore , studies of the intranuclear colocalization of the Q640X AR with cofactors , such as CBP , Q9Y3R0 , and c-Jun , reveal that the transcriptional complex that forms around the mutant AR is different to that formed around the wild type AR . We demonstrated that CBP and c-Jun are highly recruited by the mutant AR , and this leads to an unexpected activation of AP-1 , NFAT , and NFkappaB transcriptional activities . Similar enhanced activities of these transcription factors were not observed with the wild type AR . The importance of the LBD/AF-2 for the regulation of AR transcriptional activities , the impact of the presence of such AR variants on PC cells proliferation and survival , and on progression to androgen independence are discussed . Role of nitrative and oxidative DNA damage in inflammation-related carcinogenesis . Chronic inflammation induced by biological , chemical , and physical factors has been found to be associated with the increased risk of cancer in various organs . We revealed that infectious agents including liver fluke , Helicobacter pylori , and human papilloma virus and noninfectious agents such as asbestos fiber induced P35228 -dependent formation of 8-nitroguanine and 8-oxo-7 , 8-dihydro-2'-deoxyguanosine ( 8-oxodG ) in cancer tissues and precancerous regions . Our results with the colocalization of phosphorylated Q13315 and γ- P16104 with 8-oxodG and 8-nitroguanine in inflammation-related cancer tissues suggest that DNA base damage leads to double-stranded breaks . It is interesting from the aspect of genetic instability . We also demonstrated P05231 -modulated P35228 expression via P40763 and P00533 in Epstein-Barr-virus-associated nasopharyngeal carcinoma and found promoter hypermethylation in several tumor suppressor genes . Such epigenetic alteration may occur by controlling the DNA methylation through P05231 -mediated JAK/ P40763 pathways . Collectively , 8-nitroguanine would be a useful biomarker for predicting the risk of inflammation-related cancers . Role of the P08908 receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8-hydroxy-DPAT ( 8-OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 receptor interactions with epidermal growth factor ( P01133 ) . Behaviorally , the animals were more anxious . Animals treated from P01160 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin 's influence in brain development . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . Epidermal growth factor enhances androgen receptor‑mediated bladder cancer progression and invasion via potentiation of AR transactivation . P10275 ( AR ) plays a critical role in bladder cancer ( BCa ) development . Our early studies found AR knock-out mice ( with few androgens and deleted AR ) failed to develop BCa , yet 50 % of castrated mice ( with few androgens and existing AR ) still developed BCa in an N-butyl-N-(4-hydroxybutyl)nitrosamine ( BBN ) carcinogen-induced BCa mouse model , suggesting the existing AR in BCa of castrated mice may still play important roles in promoting BCa development at the castration level of androgens . The mechanism underlying this and/or which factors potentiate AR function at the castration level of androgen remains unclear . Epidermal growth factor ( P01133 ) , a key player in BCa progression , has been demonstrated to be able to potentiate AR transactivation in prostate cancer . In the present study , we found that P01133 could increase BCa cell growth , migration and invasion in the presence of AR under the low amount of androgen and P01133 was able to potentiate AR transactivation through P00533 by activating PI3K/AKT and MAPK pathway at castration androgen level . The increased suppression effects by P00533 inhibitor of PD168393 on AR function after addition of anti-androgen , DB01128 , further suggested AR might play a key role in the effects of P01133 on BCa progression and metastasis . Collectively , our results indicate that P01133 may be able to potentiate AR transactivation that leads to enhancing BCa progression , which may help us to develop a better therapeutic approach to treat BCa via targeting both P01133 and AR signaling . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Serotonin increases P27361 /2 phosphorylation in astrocytes by stimulation of P41595 and P28335 receptors . We have previously shown that fluoxetine causes P29323 (1/2) phosphorylation in cultured mouse astrocytes mediated exclusively by stimulation of 5-HT(2B) receptors ( Li et al. , 2008b ) . This raises the question whether this is also the case for serotonin ( 5-HT ) itself . In the present study serotonin was found to induce P29323 (1/2) phosphorylation by stimulation of 5-HT(2B) receptors with high affinity ( EC(50) : 20-30 pM ) , and by stimulation of 5-HT(2C) receptor with low affinity ( EC(50) : 1 microM or higher ) . P29323 (1/2) phosphorylation induced by stimulation of either 5-HT(2B) or 5-HT(2C) receptors was mediated by epidermal growth factor ( P01133 ) receptor transactivation ( Peng et al. , this issue ) , shown by the inhibitory effect of AG1478 , an inhibitor of the P01133 receptor tyrosine kinase , and DB02255 , an inhibitor of Zn-dependent metalloproteinases , and thus of 5-HT(2B) receptor-mediated P01133 receptor agonist release . It is discussed that the high potency of the 5-HT(2B)-mediated effect is consistent with literature data for binding affinity of serotonin to cloned human 5-HT(2B) receptors and with observations of low extracellular concentrations of serotonin in brain , which would allow a demonstrated moderate and modality-dependent increase in specific brain areas to activate 5-HT(2B) receptors . In contrast the relevance of the observed 5-HT(2C) receptors on astrocytes is questioned .

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interacts_with DB01257?

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Severe atypical HUS caused by P08603 S1191L -- case presentation and review of treatment options . Atypical hemolytic uremic syndrome ( aHUS ) has been associated with defective regulation of the alternative complement pathway . Although the use of plasma therapy is recommended , there is little consensus on the optimal treatment regimen . The outcome in many cases remains poor despite an improvement in our understanding of the pathology of aHUS . We have followed a female patient with aHUS associated with heterozygous complement Factor H ( P08603 ) mutation ( S1191L ) over a period of 15 years . She has been plasma dependent since infancy and has subsequently progressed to end stage kidney disease ( ESKD ) requiring dialysis treatment . Despite ESKD she still depends on regular plasma infusions to prevent thrombocytopenia . The long-term treatment plan for this patient is challenging . Renal transplantation in patients with the S1191L mutation of the P08603 gene carries a high risk of failure due to recurrence of aHUS in the renal graft . Thus , the only available curative treatment seems to be combined liver-kidney transplantation , covered by intensive plasma therapy , which comes with a high risk of morbidity and mortality . Antibodies against key activating components of the complement cascade may provide a promising alternative therapeutic strategy in the future . DB01257 , a monoclonal humanized anti- P01031 antibody , has recently been shown to be effective and well-tolerated in patients with paroxysmal nocturnal hemoglobinuria by preventing complement-mediated lysis of affected erythrocytes . Treatment of our patient with eculizumab is supported by recent reports on its successful use in two ( pediatric and adult ) patients with complement-based aHUS . P04035 inhibitors reduce interleukin-6 synthesis in human vascular smooth muscle cells . P05231 ( P05231 ) is a key molecule in chronic inflammation and has been implicated in the progression of atherosclerosis . P04035 inhibitors ( statins ) may reduce the cardiovascular risk and vulnerability of atherosclerotic plaque through nonlipid as well as lipid-lowering mechanisms , but their anti-inflammatory effects on the vascular tissue have not been fully elucidated . We investigated the effects of fluvastatin on P05231 synthesis in human vascular smooth muscle cells ( VSMCs ) . Addition of fluvastatin decreased P05231 synthesis in VSMCs in a time ( 0-24 hours ) - and dose ( 10(-8)-10(-5) mol/L ) -dependent manner . DB01095 also decreased P05231 mRNA expression in VSMCs . The effects of fluvastatin on P05231 expression were completely reversed in the presence of mevalonate or geranylgeranyl-pyrophosphate , but not squalene . Inhibition of Rho by P01024 exoenzyme or Rho kinase by Y-27632 significantly decreased P05231 expression in VSMCs . In conclusion , fluvastatin decreases P05231 synthesis in human VSMCs through inhibition of Rho pathway . These findings suggested that reduction of P05231 expression by statins may partially explain their therapeutic effects in patients with coronary artery disease . Perisurgical induction of eculizumab in a patient with paroxysmal nocturnal hemoglobinuria : its inhibition of surgery-triggered hemolysis and the consequence of subsequent discontinuation . Paroxysmal nocturnal hemoglobinuria ( PNH ) is characterized by complement ( C ' ) -induced lysis of PNH red blood cells ( RBCs ) , which are deficient in the expression of P08174 and P13987 . Surgery is one of the major clinical situations that trigger hemolytic attack and thrombosis in PNH . We describe here a case of 64-year-old man with classic PNH complicated by early-stage gastric cancer requiring distal gastrectomy under general anesthesia . We administered humanized monoclonal anti- P01031 antibody ( eculizumab ; Soliris ) for a limited period ( 600 mg , once a week × four times ) perisurgically . DB01257 effectively inhibited the C ' system and the patient underwent a curative distal gastrectomy without significant surgery-triggered hemolytic attack . Although discontinuation of eculizumab induced mild hemolysis 2 weeks after the last administration , it was treated conservatively without thrombotic complication . Limited-term induction of eculizumab could be an option for PNH patients with transient and anticipated high risks , with careful preparation for the discontinuation-related risks afterwards . DB01257 in acute recurrence of thrombotic microangiopathy after renal transplantation . Renal thrombotic microangiopathy ( TMA ) is a severe complication of systemic lupus erythematosus ( SLE ) , which is associated with the presence of antiphospholipid ( aPL ) antibodies . In its most fulminant form , TMA leads to a rapid and irreversible end-stage renal failure . DB01257 , an anti- P01031 monoclonal antibody , is a novel therapy of choice for patients with paroxysmal nocturnal hemoglobinuria ( PNH ) and atypical hemolytic uremic syndrome . Here , we report the case of a 27-year-old woman , known for SLE and end-stage renal disease due to fulminant TMA . Both aPL antibodies and antinucleosome antibodies were positive . The patient underwent a living-related kidney transplantation with immediate production of urine . Although serum creatinine was remaining high , a graft biopsy , performed on day 6 , demonstrated a TMA recurrence . Despite a treatment with plasma exchange , the situation got worse and dialysis was started . DB01257 treatment was subsequently administered and renal function improved rapidly . Three months after transplantation , serum creatinine was at 100 μmol/L , without proteinuria . This case illustrates the benefit of eculizumab therapy in a fulminant recurrence of TMA after kidney transplantation , resistant to classical therapy . The preferential beta3-adrenoceptor agonist O95696 37344 increases force via beta1-/beta2-adrenoceptors and induces endothelial nitric oxide synthase via beta3-adrenoceptors in human atrial myocardium . 1 The present study investigated the effects of the preferential beta(3)-AR agonist O95696 37344 ( O95696 ) on force of contraction ( FOC ) , Ca(2+)-transient and P29474 -activity in human right atrial myocardium . 2 O95696 concentration-dependently caused an increase in FOC that was paralleled by an increase in Ca(2+)-transient and a shortening of time to half peak relaxation ( T0.5T ) . These effects were abolished in the presence of propranolol ( 0.3 micro M ) . 3 O95696 acted as a competitive antagonist towards isoprenaline and in binding experiments it was shown to have a distinct affinity towards beta(1/2)-AR . 4 In immunohistochemical experiments O95696 ( 10 micro M ) increased detection of activated P29474 . This effect remained constant in the presence of propranolol ( 0.3 micro M ) . 5 O95696 increased directly detected NO in P08174 -staining experiments . This increase was significantly smaller in the presence of the NO-inhibitor L-NAME . 6 The inotropic effects of O95696 were not changed in the presence of L- Q13145 . 7 These results suggest that the inotropic effects of O95696 in human atrium are mediated via beta(1/2)-AR , whereas the increase of atrial P29474 -activity is due to beta(3)- adrenergic stimulation . This increase in P29474 -activity did not influence atrial myocardial contractility . In conclusion , this study shows that beta(3)-adrenergic stimulation is present in human atrium , but may not be functionally as significant as in the left ventricle . Managing a pregnant patient with paroxysmal nocturnal hemoglobinuria in the era of eculizumab . Paroxysmal nocturnal hemoglobinuria ( PNH ) is a rare clonal stem cell disorder , which affects women of child-bearing age . PNH is associated with thrombotic complications , which are the main causes of morbidity and mortality . Management of a pregnant woman with PNH remains a challenge due to high incidence of thrombotic complications and the difficulty of differentiating a PNH crisis from the complications of pregnancy . PNH is associated with an increased rate of premature labor and fetal loss . DB01257 , a humanized monoclonal antibody directed against the terminal complement protein P01031 , has revolutionized treatment of PNH . However , the role of eculizumab in pregnancy is unclear . We review the current strategies for the management of pregnant women with PNH , underline the controversies and present our recommendations . Ligands of peroxisome proliferator-activated receptor inhibit homocysteine-induced DNA methylation of inducible nitric oxide synthase gene . Homocysteine ( Hcy ) is a risk factor for atherosclerosis . It is generally accepted that inducible nitric oxide synthase ( P35228 ) is a key enzyme in the regulation of vascular disease . The aim of the present study is to investigate the effects of peroxisome proliferator-activated receptor ligands on P35228 in the presence of Hcy in human monocytes . Foam cells , induced by oxidize low density lipoprotein ( ox-LDL ) and phorbol myristate acetate ( PMA ) in the presence of different concentrations of Hcy , clofibrate and pioglitazone in human monocytes for 4 d , were examined by oil red O staining . The activity of P35228 was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot analysis . The capability of DNA methylation was measured by assaying endogenous P01031 DNA methyltransferase ( C5MTase ) activity , and the P35228 promoter methylation level was determined by quantitative MethyLight assays . The results indicated that Hcy increased the activity of C5MTase and the level of P35228 gene DNA methylation , resulting in a decrease of P35228 expression . Clofibrate and pioglitazone could antagonize the hcy effect on P35228 expression through DNA methylation , resulting in attenuation of P35228 transcription . These findings suggested that Hcy decreased the expression of P35228 by elevating P35228 DNA methylation levels , which can repress the transcription of some genes . Q07869 /gamma ligands can down-regulate P35228 DNA methylation , and could be useful for preventing Hcy-induced atherosclerosis by repressing P35228 expression . The use of the complement inhibitor eculizumab ( Soliris® ) for treating Korean patients with paroxysmal nocturnal hemoglobinuria . BACKGROUND : Paroxysmal nocturnal hemoglobinuria ( PNH ) is an acquired clonal disorder characterized by chronic complement-mediated hemolysis . DB01257 , a humanized monoclonal antibody against the terminal complement protein P01031 , potently reduces chronic intravascular hemolysis . We tested the clinical efficacy and safety of a 24-week treatment with eculizumab in 6 Korean patients with PNH . METHODS : We enrolled 6 patients with PNH who had clinically significant hemolysis . DB01257 was administered intravenously at 600 mg/week for the first 4 weeks followed by 900 mg at week 5 and 2nd weekly thereafter . RESULTS : Three men and 3 women with a median age of 39.5 years ( 24-61 years ) were enrolled . The median duration of PNH was 11 years ( 6-25 years ) . Hemolysis occurred in all patients [ median lactate dehydrogenase ( LDH ) level , 7.95 times the upper limit of the reference range of LDH ] . All patients treated with eculizumab had a rapid and sustained reduction in the degree of hemolysis . RBC transfusion requirements for 3 months were decreased from 0-12 units ( median requirement , 1.5 units ) to 0-6 units ( median requirement , 0 units ) . Improvement in fatigue was noted in 4 patients . Further , 5 patients who had been receiving corticosteroids either reduced the dose or discontinued therapy . No significant adverse events related to eculizumab therapy were observed . CONCLUSION : These results show that eculizumab reduces the degree of intravascular hemolysis , reduces or eliminates the requirement of RBC transfusion , and improves anemia and fatigue in patients with PNH . DB01257 is an effective and safe option for treating Korean patients with PNH . Cucurbitacin Q : a selective P40763 activation inhibitor with potent antitumor activity . Constitutive activation of the JAK/ P40763 pathway is a major contributor to oncogenesis . In the present study , structure-activity relationship ( SAR ) studies with five cucurbitacin ( Cuc ) analogs , A , B , E , I , and Q , led to the discovery of Cuc Q , which inhibits the activation of P40763 but not O60674 ; Cuc A which inhibits O60674 but not P40763 activation ; and Cuc B , E , and I , which inhibit the activation of both . Furthermore , these SAR studies demonstrated that conversion of the P01024 carbonyl of the cucurbitacins to a hydroxyl results in loss of anti- O60674 activity , whereas addition of a hydroxyl group to P10144 of the cucurbitacins results in loss of anti- P40763 activity . Cuc Q inhibits selectively the activation of P40763 and induces apoptosis without inhibition of O60674 , Src , Akt , Erk , or JNK activation . Furthermore , Cuc Q induces apoptosis more potently in human and murine tumors that contain constitutively activated P40763 ( i.e. , A549 , MDA-MB-435 , and v-Src/NIH 3T3 ) as compared to those that do not ( i.e. , H-Ras/NIH 3T3 , MDA-MB-453 , and NIH 3T3 cells ) . Finally , in a nude mouse tumor xenograft model , Cuc Q , but not Cuc A , suppresses tumor growth indicating that O60674 inhibition is not sufficient to inhibit tumor growth and suggesting that the ability of Cuc Q to inhibit tumor growth is related to its anti- P40763 activity . These studies further validate P40763 as a drug discovery target and provide evidence that pharmacological agents that can selectively reduce the P- P40763 levels in human cancer cells result in tumor apoptosis and growth inhibition . [ Paroxysmal nocturnal hemoglobinuria therapy with eculizumab : Spanish experience ] . BACKGROUND AND OBJECTIVES : Paroxysmal nocturnal hemoglobinuria ( PNH ) is a rare acquired clonal disease characterized by complement-mediated hemolysis , bone marrow failure and thrombosis . DB01257 is a humanized monoclonal antibody that blocks the cytolytic component of the complement system by binding to complement P01031 . MATERIAL AND METHODS : We report the results of eculizumab treatment in 25 PNH patients from different centers in Spain . Statistical analysis was perfomed with a SPSS v15.0 software . RESULTS : Fifty-eight per cent of the patients achieved transfusional independence after a median of 14 months . Transfusion requirements were reduced in 60 % of the remaining cases . Fatigue resolved in 96 % of the patients and smooth muscle dystony-related symptoms in all cases . A single case of treatment-related infection was observed . CONCLUSIONS : DB01257 controls effectively hemolysis and greatly improves clinical symptoms . The drug is safe and well tolerated , without significant adverse effects except meningococcal infection . Patients with suboptimal response to treatment must be assessed for bone marrow insufficiency and extravascular haemolysis . Conformationally constrained DB05875 analogues : the total synthesis of a constrained peptidomimetic for the Phe7-Phe8 region . A lactam-based peptidomimetic for the Phe7-Phe8 region of DB05875 has been synthesized . The synthesis used an anodic amide oxidation to selectively functionalize the P01031 -position of a 3-phenylproline derivative . The resulting proline derivative was coupled to a Cbz-protected phenylalanine , and an intramolecular reductive amination strategy used to convert the coupled material into a bicyclic piperazinone ring skeleton . The net result was a dipeptide building block that imbedded one of two proposed receptor bound conformations for the Phe7-Phe8 region of DB05875 into a bicyclic ring skeleton . The building block was then converted into a constrained DB05875 analogue with the use of solid-phase peptide synthesis . A similar intramolecular reductive amination strategy was used to synthesize a DB05875 analogue having only Phe7 constrained , and the original 3-phenylproline was converted into a DB05875 analogue having only Phe8 constrained . All of the analogues were examined for their ability to displace DB05875 from its P25103 . [ Clinical aspects of the complement system ] . The complement system consists of more than 30 proteins and has 3 types of activation pathways : classical , lectin and alternative pathways . The complement system not only has a role in innate immunity but also works as an antibody-dependent effecter to eliminate pathogens . It is useful to measure serum levels of CH50 , P01024 and C4 in patients with immune-mediated diseases . While increased levels of CH50 are associated with non-specific inflammation , decreased levels of CH50 in combination with normal or decreased levels of P01024 and C4 are associated with specific immune-mediated diseases . Recent studies have demonstrated that the defect in the clearance of immune complexes and apoptotic cells is associated with autoimmune disease . Mice deficient in Clq show a lupus-like phenotype with the appearance of antinuclear antibodies and glomerulonephritis due to a defect in the clearance of immune complexes and apoptotic cells . This at least explains the paradox that , in humans , deficiency in an early complement component is a major risk factor for SLE . It is demonstrated that mutations in factor H , membrane cofactor protein ( MCP ) and factor I gene are associated with atypical hemolytic uremic syndrome . Since the complement system is a central mediator of inflammation , it is recognized as a promising therapeutic target . Anti- P01031 monoclonal antibody was developed to block the final stage of complement activation . DB04949 is a single chain , short-acting anti- P01031 antibody and is used for reperfusion after myocardial infarction , or for coronary artery bypass graft surgery with cardiopulmonary bypass . DB01257 is a long-acting anti- P01031 antibody used for paroxysmal nocturnal hemoglobinuria , rheumatoid arthritis , membranous glomerulonephritis with promising results . N-aryl-N'-benzylpiperazines as potential antipsychotic agents . N1-(2-Alkoxyphenyl)piperazines additionally containing an N4-benzyl group bearing alcohol , amide , imide , or hydantoin functionalities were prepared and evaluated in the conditioned avoidance response ( CAR ) test predictive of clinical antipsychotic activity and in in vitro receptor-binding assays . Certain of the compounds display high affinity for the D2 , P08908 , and alpha 1-adrenergic receptors . Structures bearing acyclic amide , lactam , and imide functionalities display good biological activity , with a preference for the 1,3-disubstituted phenyl ring relative to the 1,4- and 1,2-congeners ( 7 vs 10 and 12 ) . Every possible position of hydantoin attachment was investigated ( e.g. , substitution at N1 , N3 , and P01031 ) . The hydantoin involving attachment to N1 ( 24 ) was found to have good biological activity , whereas those hydantoins with attachment to N3 or P01031 ( 22 , 23 , and 25 ) were inactive . Several of the smaller acetylated derivatives ( 30 and 33 ) have fair in vivo activity , which was lost in the case of the larger benzoyl analog 31 . DB03419 congener 34 had modest affinity for the D2 receptor ( 65 nM ) as well as excellent in vivo activity . Benzylamino compounds display ( viz. 27 and 35-38 ) moderate CAR activity but have surprising receptor affinity , often greater than those of comparable structures bearing a carbonyl ( 36 vs 7 ) . Benzyl and benzhydryl alcohol compounds 40-48 are more active than amino structures 27 and 35-38 and also exhibit excellent in vivo activity in the CAR test with modest D2 and P08908 receptor binding . DB01257 in atypical haemolytic-uraemic syndrome allows cessation of plasma exchange and dialysis . Disorders in complement regulation are a major cause of atypical haemolytic-uraemic syndrome ( aHUS ) . DB01257 , a monoclonal antibody targeting complement P01031 and blocking the terminal complement cascade , should theoretically be useful in this disease , particularly when associated with specific complement pathway anomalies such as Factor H deficiency . DB01257 is emerging as an effective treatment for post-transplant aHUS recurrence and may have a role in treating de novo aHUS , halting the haemolytic process . In this case report , we describe the fourth case of aHUS treated with eculizumab . In our patient , with a known complement Factor H mutation , not only has the disease process become quiescent but also this therapy has led to significantly improved renal function so that dialysis is no longer necessary . The role of complement in atherosclerosis . PURPOSE OF REVIEW : Although it has long been recognized that atherosclerotic lesions show evidence of complement activation , the functional roles of the complement system in atherogenesis are not yet fully resolved . This article highlights recent publications on the complement system in the atherosclerosis field . RECENT FINDINGS : There have been a number of recent papers reporting on the association of complement proteins and complement regulators with high density lipoproteins , complement activation by enzymatically-modified LDL , signalling pathways downstream of C3a and C5a receptors and membrane C5b-9 assembly , and the prevention of C5b-9 assembly on endothelial cells via upregulation of P13987 expression in response to arterial laminar flow . C1q has been found to play a protective role in early lesion formation in P01130 deficient mice , and Crry-Ig and soluble C1 inhibitor have both been shown to have therapeutic effects in models of vascular injury in ApoE deficient mice . The possibility that the Y402H Factor H polymorphism influences atherosclerosis has been supported in a recent paper showing increased risk in white hypertensive individuals . SUMMARY : The articles that have emerged over the last year highlight the relevance of the complement system to the atherosclerosis field . DB00277 treatment improves mitochondrial function after upper cervical spinal cord hemisection . The importance of mitochondria in spinal cord injury has mainly been attributed to their participation in apoptosis at the site of injury . But another aspect of mitochondrial function is the generation of more than 90 % of cellular energy in the form of DB00171 , mediated by the oxidative phosphorylation ( OxPhos ) process . P99999 oxidase ( CcO ) is a central OxPhos component and changes in its activity reflect changes in energy demand . A recent study suggests that respiratory muscle function in chronic obstructive pulmonary disease ( P48444 ) patients is compromised via alterations in mitochondrial function . In an animal model of cervical spinal cord hemisection ( C2HS ) respiratory dysfunction , we have shown that theophylline improves respiratory function . In the present study , we tested the hypothesis that theophylline improves respiratory function at the cellular level via improved mitochondrial function in the C2HS model . We demonstrate that CcO activity was significantly ( 33 % ) increased in the spinal cord adjacent to the site of injury ( P01024 - P01031 ) , and that administration of theophylline ( 20mg/kg 3x daily orally ) after C2HS leads to an even more pronounced increase in CcO activity of 62 % compared to sham-operated animals . These results are paralleled by a significant increase in cellular DB00171 levels ( 51 % in the hemidiaphragm ipsilateral to the hemisection ) . We conclude that C2HS increases energy demand and activates mitochondrial respiration , and that theophylline treatment improves energy levels through activation of the mitochondrial OxPhos process to provide energy for tissue repair and functional recovery after paralysis in the C2HS model . DB01257 therapy for atypical haemolytic uraemic syndrome due to a gain-of-function mutation of complement factor B . BACKGROUND : Atypical haemolytic uraemic syndrome ( aHUS ) is caused by dysregulated complement activation . A humanised anti- P01031 monoclonal antibody has recently become available for treatment of this condition CASE-DIAGNOSIS/TREATMENT : We present the first description of an infant with an activating mutation of complement factor B successfully treated with eculizumab . On standard doses she had evidence of ongoing P01031 cleavage despite a good clinical response . CONCLUSIONS : DB01257 is effective therapy for aHUS associated with factor B mutations , but recommended doses may not be adequate for all patients . Role of monoclonal antibodies in the treatment of immune-mediated glomerular diseases . Non-specific immunosuppressants have represented for decades the only therapies for patients with immune-mediated glomerular diseases . These treatments , however , are associated with high rates of no-response and are burdened by toxicities that frequently offset the benefits of proteinuria reduction . Monoclonal antibodies targeting selective cell populations or mediators implicated in the pathophysiology of glomerular diseases have recently become available . DB00073 , a chimeric monoclonal antibody against the P11836 antigen on B cells , safely reduced proteinuria in patients with nephrotic syndrome secondary to membranous nephropathy , minimal change disease , or focal segmental glomerulosclerosis . Its ability to reduce auto-antibody formation has been instrumental to treat also ANCA-associated vasculitis , lupus nephritis , and mixed cryoglobulinemia . Many reports have also documented the efficacy of the anti- P01031 humanized monoclonal antibody DB01257 to treat atypical hemolytic uremic syndrome , P01024 nephropathy , and membranoproliferative glomerulonephritis . Thanks to these encouraging findings , monoclonals are becoming very helpful tools to treat patients with glomerular diseases . Moreover , thanks to their specific mechanism of action , these and other monoclonal antibodies are important in improving our understanding of the pathophysiology of glomerular diseases . Their still high costs , however , might represent a major hurdle for their widespread implementation for all patients in need . [ P01031 : eculizumab ] . DB01257 is a recombinant humanized monoclonal antibody that specifically binds to a P01031 terminal complement and inhibits the cleavage of P01031 to C5a and C5b through complement activation . DB01257 has been used clinically for paroxysmal nocturnal hemoglobinuria . Clinical studies have been initiated for neurological disorders , such as , optic neuromyelitis and myasthenia gravis , mediated by the complement system . In addition , eculizumab is expected to be useful for the treatment of intractable neurological diseases . Angiotensin-converting-enzyme inhibitors suppress synthesis of tumour necrosis factor and interleukin 1 by human peripheral blood mononuclear cells . Administration of angiotensin-converting-enzyme ( P12821 ) inhibitors reduce vascular proliferation following endothelial injury as well as progression of renal disease in various animal models . These effects might be due to interference with cytokines such as interleukin 1 ( IL-1 ) or tumour necrosis factor alpha ( P01375 ) since they have been implicated in regulating the effects of vascular cell growth factors such as fibroblast- and platelet-derived growth factors . We investigated the in vitro synthesis of IL-1 and P01375 from human peripheral blood mononuclear cells ( PBMC ) in the presence of various P12821 -inhibitors . DB01197 dose-dependently suppressed the P01584 -induced synthesis of P01375 by 74 % ( P < 0.01 ) and the P01584 -induced synthesis of P01583 by 60 % ( P < 0.01 ) . Cytokine synthesis induced by lipopolysaccharide was less affected . At concentrations suppressing P01375 and IL-1 , captopril did not reduce the synthesis of complement P01024 in the same cells . Enalapril and cilazapril also suppressed cytokine-induced cytokine synthesis . Ramipril , lisinopril , perindopril and spirapril had no significant effect on P01375 synthesis suggesting that the effect was not related specifically to the inhibition of P12821 . Accumulation of mRNA for IL-1 and P01375 were not affected by captopril , suggesting a posttranscriptional effect . We conclude that certain P12821 -inhibitors suppress IL-1 and P01375 synthesis at a posttranscriptional level and might therefore influence cytokine-mediated cell growth . Complement fraction 3 binding on erythrocytes as additional mechanism of disease in paroxysmal nocturnal hemoglobinuria patients treated by eculizumab . In paroxysmal nocturnal hemoglobinuria ( PNH ) hemolytic anemia is due mainly to deficiency of the complement regulator P13987 on the surface of red blood cells ( RBCs ) . DB01257 , an antibody that targets complement fraction 5 ( P01031 ) , has proven highly effective in abolishing complement-mediated intravascular hemolysis in PNH ; however , the hematologic benefit varies considerably among patients . In the aim to understand the basis for this variable response , we have investigated by flow cytometry the binding of complement fraction 3 ( P01024 ) on RBCs from PNH patients before and during eculizumab treatment . There was no evidence of P01024 on RBCs of untreated PNH patients ; by contrast , in all patients on eculizumab ( n = 41 ) a substantial fraction of RBCs had P01024 bound on their surface , and this was entirely restricted to RBCs with the PNH phenotype ( P13987 (-) ) . The proportion of P01024 (+) RBCs correlated significantly with the reticulocyte count and with the hematologic response to eculizumab . In 3 patients in whom (51)Cr labeling of RBCs was carried out while on eculizumab , we have demonstrated reduced RBC half-life in vivo , with excess (51)Cr uptake in spleen and in liver . Binding of P01024 by PNH RBCs may constitute an additional disease mechanism in PNH , strongly enhanced by eculizumab treatment and producing a variable degree of extravascular hemolysis . Timing of eculizumab therapy for P01024 glomerulonephritis . DB01257 is an anti- P01031 antibody that inhibits P01031 cleavage and prevents the generation of the terminal complement complex C5b-9 . DB01257 is licensed to treat paroxysmal nocturnal haemoglobinuria or atypical haemolytic uraemic syndrome ( aHUS ) . Clinical trials are ongoing for P01024 glomerulopathy . Given the unfamiliarity of physicians with these rare diseases and the variability of clinical presentation , a delayed initiation of eculizumab therapy is common . Thus , the question arises as to what extent improvement of kidney function may be expected when patients have been dialysis dependent for weeks or months already when eculizumab is initiated . Furthermore , given the high cost and potential adverse effects of eculizumab , the question arises of when to stop therapy because of futility when patients with kidney-only manifestations remain dialysis dependent . In literature reports , eculizumab was stopped as early as after 3 weeks because the patient remained dialysis dependent . In this issue of CKJ , Inman et al. report on eculizumab-induced reversal of dialysis-dependent kidney failure from P01024 glomerulonephritis , illustrating both the potential benefit of eculizumab for this complement-mediated disease and the need for lengthy therapy-dialysis independency was reached after 5 months of eculizumab . Indeed , there are reports of renal function recovery when eculizumab was initiated after 4 months on dialysis and of recovery of renal function 2.0-3.5 months after initiation of eculizumab in dialysis-dependent patients with P01024 glomerulopathy or aHUS . Drugs that inhibit complement . The complement system is an important part of the innate immune system . Complement plays a crucial role in the pathophysiology of many disorders . Despite the pivotal role of the complement system , an approved targeted inhibitor of a complement factor became available only recently . DB01257 is a humanized monoclonal antibody that inhibits complement factor P01031 . It is a targeted , disease modifying , treatment of paroxysmal nocturnal hemoglobinuria ( PNH ) . It was approved be the US FDA and the European Commission in 2007 . In this review we will update the experience with eculizumab in PNH and discuss potential use of eculizumab in other disorders ( e.g. cold agglutinin disease ; atypical HUS ) and new approaches to complement inhibition with drugs other than eculizumab . Noninvasive detection of complement activation through radiologic imaging . A wealth of experimental and clinical data demonstrates that the complement system is involved in the pathogenesis of numerous inflammatory diseases . Complement activation contributes to injury in disorders that involve nearly every tissue in the body . Concerted effort has been expended in recent years to develop therapeutic complement inhibitors . DB01257 , an inhibitory antibody to P01031 , was recently approved for the treatment of several diseases , and many other complement inhibitors are in clinical development . As these drugs are developed , the need for improved methods of detecting and monitoring complement activation within particular tissues will be increasingly important . We have developed a magnetic resonance imaging ( Q9BWK5 ) -based method for noninvasive detection of complement activation . This method utilizes iron-oxide nanoparticles that are targeted to sites of complement activation with a recombinant protein that contains the C3d-binding region of complement receptor ( CR ) 2 . DB01592 -oxide nanoparticles darken ( negatively enhance ) images obtained by P24752 -weighted Q9BWK5 . We have demonstrated that the P20023 -targeted nanoparticles bind within the kidneys of mice with lupus-like kidney disease ( MRL/1pr mice ) , causing a decrease in the P24752 signal within the kidneys . This method discriminates diseased kidneys from healthy controls , and the magnitude of the negative enhancement in the cortex of MRL/lpr mice correlates with their disease severity . This method may be useful for identifying those patients most likely to benefit from complement inhibitors and for monitoring the response of these patients to treatment . These results may open up new avenues to develop tools for the monitoring of disease progression in complement-dependent diseases . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . A P04035 inhibitor possesses a potent anti-atherosclerotic effect other than serum lipid lowering effects -- the relevance of endothelial nitric oxide synthase and superoxide anion scavenging action . We have determined whether the anti-atherosclerotic effect of a 3-hydroxy-3-methyl-glutaryl- DB01992 ( HMG- DB01992 ) reductase inhibitor ( fluvastatin ) is mediated through nitric oxide ( NO ) as well as affecting plasma lipids . NO related vascular responses , endothelial nitric oxide synthase ( P29474 ) mRNA and superoxide anion ( O(2)(-) ) release were examined in vascular walls of oophorectomized female rabbits fed 0.5 % cholesterol chow for 12 weeks with or without fluvastatin ( 2 mg/kg per day ) . Serum lipid profile was not different between two groups . NO dependent responses stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-L-arginine acetate ( L- Q13145 ) ; nitric oxide synthase inhibitor were all improved by fluvastatin treatment . Endothelium independent vasorelaxation induced by nitroglycerin was not different between the two groups of rabbits ' arteries . DB01095 treatment increased cyclic GMP concentration in aorta of rabbits . P29474 mRNA expression and O(2)(-) release were measured in aorta using competitive reverse transcription-polymerase chain reaction ( RT-PCR ) and with lucigenin analogue , 2-methyl-3,7-dihydroimidazol [1,2-a]pyrazine-3-one ( MCLA ) chemiluminescence methods . P29474 mRNA in the endothelial cells of aorta was significantly up-regulated and O(2)(-) production was significantly reduced in fluvastatin treated rabbit aorta . Anti-macrophage staining area , but not anti-smooth muscle cell derived actin stained area in the aorta was also reduced by fluvastatin treatment . Conclusion , fluvastatin , a P04035 inhibitor , retards the initiation of atherosclerosis formation through the improvement of NO bioavailability by both up-regulation of P29474 mRNA and decrease of O(2)(-) production in vascular endothelial cells , and this means that part of the anti-atherosclerotic effect of fluvastatin may be due to nonlipid factors . Dissecting the complement pathway in hepatic injury and regeneration with a novel protective strategy . Liver resection is commonly performed under ischemic conditions , resulting in two types of insult to the remnant liver : ischemia reperfusion injury ( IRI ) and loss of liver mass . Complement inhibition is recognized as a potential therapeutic modality for IRI , but early complement activation products are also essential for liver regeneration . We describe a novel site-targeted murine complement inhibitor , P20023 - P13987 , which specifically inhibits the terminal membrane attack complex ( MAC ) , and we use this protein to investigate the complement-dependent balance between liver injury and regeneration in a clinical setting of pharmacological inhibition . P20023 - P13987 did not impact in vivo generation of P01024 and P01031 activation products but was as effective as the P01024 activation inhibitor P20023 -Crry at ameliorating hepatic IRI , indicating that the MAC is the principle mediator of hepatic IRI . Furthermore , unlike P01024 or P01031 inhibition , P20023 - P13987 was not only protective but significantly enhanced hepatocyte proliferation after partial hepatectomy , including when combined with ischemia and reperfusion . Remarkably , P20023 - P13987 also enhanced regeneration after 90 % hepatectomy and improved long-term survival from 0 to 70 % . P20023 - P13987 functioned by increasing hepatic P01375 and P05231 levels with associated P40763 and Akt activation , and by preventing mitochondrial depolarization and allowing recovery of DB00171 stores . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Immunohistochemical analysis in ethinylestradiol-treated breast cancers after prior long-term estrogen-deprivation therapy . BACKGROUND : P03372 ( ER ) positive breast cancer can often be treated by hormone therapy ; however a certain population of ER-positive patients become resistant to hormone therapy after long-term hormone treatment . DB00977 ( EE2 ) is a derivative of estrogen , which has shown promising effects in these patients . METHODS : We successfully obtained tissue samples from 6 patients undergoing EE2 treatment and examined 13 well-known breast cancer-related factors by immunohistochemistry . Of the 6 patients , 5 responded but one patient did not . RESULTS : Before EE2 treatment , staining for both ER and androgen receptor ( AR ) was strong in the nucleus , and the progesterone receptor ( PgR ) was almost no staining . EE2 treatment significantly down-regulated ER and up-regulated PgR while nuclear and cytosolic AR were oppositely down- and up-regulated , respectively . Cytosolic staining of P38398 was significantly up-regulated by EE2 whereas nuclear staining tended to decrease . Individual comparisons suggested less induction of PgR and decreasing AKT but increasing pAKT in the non-responder following EE2 treatment . CONCLUSIONS : Our observations revealed that EE2 activated ER downstream genes ; however it did not stimulate cell growth . This suggests that hormone resistant cells might receive growth signals from a non-genomic pathway and this may be reflected in their sensitivity to EE2 treatment . DB01257 for paroxysmal nocturnal haemoglobinuria . The complement system plays a central part in both innate and acquired immunity , but the contribution of complement activation to pathobiology is largely ancillary . An exception to the non-dominant role of complement in disease is the haemolytic anaemia of paroxysmal nocturnal haemoglobinuria ( PNH ) . The intravascular haemolysis that is the clinical hallmark of PNH is a consequence of deficiency of the complement inhibitory proteins decay accelerating factor ( P08174 , P08174 ) and membrane inhibitor of reactive lysis ( P13987 , P13987 ) . DB01257 is a humanised monoclonal antibody that binds and prevents activation of complement P01031 and the subsequent formation of the cytolytic membrane attack complex of complement . DB01257 inhibits the intravascular haemolysis of PNH , reduces transfusion requirements , stabilises haemoglobin concentration , and improves quality of life . Although chronic treatment with eculizumab increases the risk of infections with Neisseria meningitides , the drug is generally safe and well tolerated . But as is the case with other drugs developed for treatment of ultra-orphan diseases , eculizumab is expensive , and treatment must continue indefinitely because P01031 inhibition does not affect the process ( ie , clonal proliferation of haemopoietic stem cells with a mutant phosphatidylinositol glycan complementation class A [ P37287 ] gene ) that underlies PNH . Moreover , due to the heterogeneous nature of the disease , treatment with eculizumab is not appropriate for all patients with PNH . DB01257 treatment for rescue of renal function in IgA nephropathy . BACKGROUND : Immunoglobulin A ( IgA ) nephropathy is a chronic glomerulonephritis with excessive glomerular deposition of IgA1 , P01024 and C5b-9 , which may lead to renal failure . CASE DIAGNOSIS/TREATMENT : We describe the clinical course of an adolescent with rapidly progressive disease leading to renal failure in spite of immunosuppressive treatment . Due to refractory disease the patient was treated with eculizumab ( anti- P01031 ) for 3 months in an attempt to rescue renal function . Treatment led to clinical improvement with stabilization of the glomerular filtration rate and reduced proteinuria . Discontinuation of treatment led to a rapid deterioration of renal function . This was followed by a single dose of eculizumab , which again reduced creatinine levels temporarily . CONCLUSIONS : Early initiation of eculizumab therapy in patients with progressive IgA nephropathy may have a beneficial effect by blocking complement-mediated renal inflammation . DB01257 for the treatment of preeclampsia/HELLP syndrome . Severe preeclampsia with hemolysis , elevated liver enzymes and low platelets ( HELLP ) syndrome is a leading cause of maternal and neonatal morbidity and mortality worldwide . Occurrence at an extremely premature gestational age is most challenging as there are dichotomous imperatives : delivery as definitive therapy for maternal health vs. prolongation of pregnancy to avoid prematurity and associated morbidities . We describe a patient presenting with severe preeclampsia/HELLP syndrome at 26 weeks gestation that was treated with DB01257 , a targeted inhibitor of complement protein P01031 , which resulted in marked clinical improvement and complete normalization of lab parameters . Pregnancy was prolonged 17 days , likely resulting in a reduction of neonatal morbidity with its associated short and long-term health care costs . Successful use of DB01257 in this case suggests that complement inhibition may be an effective treatment strategy for severe preeclampsia/HELLP syndrome . DB01257 treatment efficiently prevents P01031 cleavage without C5a generation in vivo . Neurokinin 1 receptor mediates membrane blebbing in HEK293 cells through a Rho/Rho-associated coiled-coil kinase-dependent mechanism . We have investigated the effect of neurokinin 1 receptor ( P25103 ) agonists on HEK293 cells transfected with the P25103 receptor . The P25103 receptor mediates dramatic shape changes that include contractions of the membrane cortex resulting in membrane bleb formation . We have found that the cell shape changes correlate with changes in electrical impedance measured in cellular monolayers . The shape and impedance changes were prevented after preincubation with P25103 antagonists aprepitant and L-73060 . Although bleb formation usually heralds apoptotic cell death , we have found that P25103 -mediated cellular blebbing does not associate with apoptosis . Preincubation with a cell-permeable derivative of P01024 transferase that blocks Rho or with the Rho-associated coiled-coil kinase inhibitor Y27632 completely prevented P25103 -induced shape and impedance changes . Blebbing was also completely inhibited by ML-9 , a myosin light chain kinase inhibitor . Furthermore , the phospholipase C inhibitor U73,122 did not interfere with the effect of Substance P ( SP ) on cellular morphology and cellular impedance but completely blocked SP-induced intracellular calcium increase , indicating that the blebbing is a process independent of intracellular calcium elevations . Blebbing is a protein kinase C-independent process , since the nonselective protein kinase C inhibitor GF109203X did not interfere with SP-induced effects . Based on these results , we provide the first evidence that P25103 receptor-ligand interaction can cause apoptosis-independent cellular blebbing and that this process is mediated by the Rho/Rho-associated coiled-coil kinase pathway . DB01257 and drug-induced haemolytic-uraemic syndrome . The monoclonal anti- P01031 antibody eculizumab has been successfully tested in atypical haemolytic-uraemic syndrome ( aHUS ) , with or without mutations in the regulatory proteins of the alternative pathway of the complement , and less convincingly in enterohaemorrhagic Escherichia coli-associated HUS . Here , we report a patient with mitomycin-C-induced HUS unresponsive to plasma exchanges . DB01257 infusion was followed by a dramatic improvement of haematological parameters and renal function , suggesting a role of complement blockade in the management of refractory , drug-related HUS . DB01257 for treatment of asthma . INTRODUCTION : Asthma is an inflammatory disease , which can be exacerbated by stimuli such as viral infections and exposure to allergens . Asthma continues to be a profound public health problem due to asthma exacerbation in a low proportion of patients in need of more effective medications . AREAS COVERED : The P01031 complement pathway has been proposed as a new target for the treatment of asthma , supported by clinical observations of increased C5a levels in asthmatic airways , constitutive expression of P01031 receptors on bronchial epithelium and smooth muscle cells , and preclinical studies in mice demonstrating inhibition of P01031 cleavage reduced established airway inflammation and improves lung function . DB01257 is a monoclonal antibody , which binds to the complement protein P01031 , thereby preventing the formation of C5a and C5b-9 . The research discussed in this review describes development of eculizumab for the treatment of paroxysmal nocturnal hemoglobinuria and the efficacy of eculizumab on allergen-induced asthmatic responses in a placebo-controlled study . EXPERT OPINION : In an allergen-challenge model of asthma , there was a significant period effect with eculizumab , with inhibition of the late asthmatic response in subjects who received placebo treatment first . Although this study provides some evidence that eculizumab may be effective to attenuate allergen-induced responses , the role of P01031 in asthma remains to be clarified . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . Tailored eculizumab therapy in the management of complement factor H-mediated atypical hemolytic uremic syndrome in an adult kidney transplant recipient : a case report . Atypical hemolytic uremic syndrome ( aHUS ) is characterized by thrombocytopenia , microangiopathic hemolytic anemia , and acute kidney injury ( AKI ) which frequently progresses to end-stage renal disease ( ESRD ) . In 50 % of affected patients , mutations in complement regulatory proteins cause inappropriate complement activation with endothelial injury . P08603 ( P08603 ) mutations cause 25 % of aHUS cases ; these patients have an 80 % recurrence risk after kidney transplantation . DB01257 , an anti- P01031 antibody , is effective in limiting hemolysis episodes in patients with aHUS , but less is known about preventing recurrence after kidney transplantation . Herein we report the use of prophylactic eculizumab in an adult with aHUS who underwent kidney transplantation . A 31-year-old female presented with aHUS and progressive AKI associated with low complement 3 level leading to ESRD despite plasmapheresis and corticosteroids . She had a heterozygous nonsense mutation in P08603 and reduced plasma P08603 levels . She was given preoperative plasmapheresis and eculizumab and underwent living unrelated renal transplantation . Postoperatively , eculizumab was dosed to achieve low functional complement 5 levels and low soluble membrane attack complex levels and she has maintained excellent graft function without aHUS recurrence . We propose that eculizumab with titrated dosing should be used in P08603 -mediated aHUS patients who are at a high risk of recurrence . Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3/D2 receptor agonists 7-OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1/D5 agonist SKF38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7-OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7-OH-DPAT > PD128907 = apomorphine ) . DB01200 and SKF38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2/D3 antagonist raclopride blocked both 7-OH-DPAT-induced hypothermia and 7-OH-DPAT-induced PPI disruption . The P08908 antagonist WAY 100,135 , and the peripheral D2-like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors . Atrial natriuretic peptides inhibit conductive sodium uptake by rabbit inner medullary collecting duct cells . The inner medullary collecting duct ( IMCD ) effects net sodium reabsorption under the control of volume regulatory hormones , including atrial natriuretic peptides ( P01160 ) . These studies examined the mechanisms of sodium transport and its regulation by P01160 in fresh suspensions of IMCD cells . Sodium uptake was inhibited by amiloride but insensitive to furosemide , bu-metanide , and hydrochlorthiazide . These results are consistent with uptake mediated by a sodium channel or Na+/H+ exchange . To determine the role of sodium channels , cells were hyperpolarized by preincubation in high potassium medium followed by dilution into potassium-free medium . Membrane potential measurements using the cyanine dye , Di(S)- P01024 -5 verified a striking hyperpolarization of IMCD cells using this protocol . Hyperpolarization increased the apparent initial rate of sodium uptake fourfold . DB00594 and P01160 inhibited potential-stimulated sodium uptake 73 % and 65 % , respectively ; the two agents together were not additive . Addition of 5 mM sodium to hyperpolarized cells resulted in a significant amiloride-sensitive depolarization . Half-maximal inhibition of potential-driven sodium uptake occurred at 3 X 10(-7) M amiloride , and 5 X 10(-11) M P01160 . We conclude that sodium enters IMCD cells via a conductive , amiloride-sensitive sodium channel , which is regulated by P01160 . P01160 inhibition of luminal sodium entry in the IMCD appears to contribute to the marked natriuretic effect of this hormone in vivo . Successful management of obstructive jaundice due to gallstones with eculizumab in a patient with paroxysmal nocturnal hemoglobinuria . Paroxysmal nocturnal hemoglobinuria ( PNH ) makes patients susceptible to intravascular hemolysis and thrombosis , and it can be life-threatening in stressful situations . DB01257 , a humanized monoclonal antibody that inhibits the complement protein P01031 , has been evaluated as a novel therapy for PNH . We herein describe the case of a 59-year-old Japanese woman with classic PNH , who had been successfully treated with eculizumab , but who later developed acute cholecystitis/cholangitis from gallstones . Although the severe obstructive jaundice requiring endoscopic therapy following cholecystectomy was complicated , critical intravascular hemolysis and thrombosis were not observed . Therefore , utilizing eculizumab during the peri-operative management of PNH patients should be carefully taken into consideration . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . Changes in the expression of plasma proteins associated with thrombosis in P38398 mutation carriers . PURPOSE : Although P38398 gene mutations have been associated with breast cancer , P38398 mutations have been also involved in other functions . Thrombosis and coagulation are novel mechanisms recently associated with cancer . The aims of the present study were ( a ) to evaluate , using proteomics , if P38398 mutation carriers have a different plasma proteins expression related to thrombosis and coagulation profile than non-mutant P38398 women and ( b ) to analyze if the expression of these proteins may be different among P38398 mutation carriers with and without breast cancer . METHODS : Proteomic study was based on 2-dimensional electrophoresis and mass spectrometry . The study was performed in 10 P38398 non-mutant controls and 21 women with P38398 mutations ( with breast cancer ( n = 8 ) and breast cancer-free ( n = 13 ) ) , all of them free of family history or diagnosis of ovarian cancer . RESULTS : Proteomic study showed that fibrinogen gamma chain isotypes 2 and 3 , serotransferrin isotype 4 , and convertase P01024 / P01031 isotypes 1-5 were significantly increased in plasma from P38398 mutation carriers with respect to P38398 non-mutant controls . Plasma levels of alpha-1 antitrypsin isotypes 2-5 , apolipoprotein A-IV , and vitamin D-binding protein isotypes 1 and 2 were significantly reduced in P38398 mutation carriers with respect to non-mutant controls . Only apolipoprotein A-IV plasma levels were significantly higher in cancer-free P38398 mutations carriers compared with P38398 mutations carriers who developed breast cancer . CONCLUSION : It is suggested that independently of breast cancer generation , P38398 -encoded gene alterations are associated with changes in the expression of circulating proteins associated with thrombosis and coagulation . Antibody-mediated rejection in kidney transplantation : an update . INTRODUCTION : Acute antibody-mediated rejection ( AMR ) in renal-transplant recipients is generally less responsive to conventional antirejection therapy and has a worse prognosis than acute cellular rejection . AREAS COVERED : This review provides a broad understanding of the pathogenesis of AMR , recent advances in its therapy , and future directions . Conventional therapeutic approaches to AMR have minimal impact on mature plasma cells , the major source of antibody production . Emerging therapies include bortezomib , a proteasome inhibitor , and eculizumab , an anti- P01031 antibody . In several reports , bortezomib therapy resulted in prompt reversal of rejection , decreased titers of donor-specific antibodies ( DSA ) , and improved renal allograft function . DB01257 also reversed AMR and prevented its development in patients with high post-transplantation DSA levels . EXPERT OPINION : Despite the small sample size and lack of controls , these studies are encouraging , and although larger studies and long-term follow-up are needed , bortezomib and eculizumab may play a major future role in AMR therapy . Mycobacteria-primed macrophages and dendritic cells induce an up-regulation of complement C5a anaphylatoxin receptor ( CD88 ) in CD3+ murine T cells . Complement C5a anaphylatoxin is a potent activator of macrophages , neutrophils , and dendritic cells ( DC ) and binds the C5a receptor ( P21730 ; CD88 ) . Although C5a is chemotactic for T cells , expression of P21730 on murine T cells has been disputed . We report here that naïve , Con A-activated , and cytokine ( IL-12 , Q14116 ) -stimulated murine CD3+ T cells from three strains of mice [ C57Bl/6 , B10.nSn ( P01031 +/+ ) , B10.on ( P01031 -/- ) ] lacked P21730 , as evaluated by immunophenotyping with an anti- P21730 mAb . Ligation of CD3 induced a modest up-regulation with 3 % of CD3+ T cells expressing cell surface P21730 . T cells primed by P25054 differentiate into effector T cells . Activation of mycobacteria [ bacillus Calmette-Guerin ( BCG ) ] -sensitized T cells through MHC II and TCR interactions via BCG-infected macrophages enhanced the expression of P21730 with approximately 14 % of CD3+ T cells positive for P21730 . Comparable expression was found in P01031 +/+ as well as P01031 -/- strains of mice ( 14 % and 15 % , respectively ) . Furthermore , anti-CD3-activated T cells were primed by BCG-infected DC , and a larger proportion of the primed T cells expressed P21730 ( 30-40 % ) . Finally , mice infected with BCG showed significant numbers of CD3+ T cells expressing P21730 in the spleens during infection . As P25054 , such as macrophages and DC , can secrete P01031 and cleave P01031 to C5a and C5b through a peptidase , we suggest that macrophage and DC-T cell interactions can up-regulate P21730 on T cells through MHC II-TCR and provide a C5a peptide for additional local activation of T cells via P21730 . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . Cross-talk between PKA-Cβ and p65 mediates synergistic induction of Q07343 by roflumilast and NTHi . Phosphodiesterase 4B ( Q07343 ) plays a key role in regulating inflammation . DB01656 , a phosphodiesterase (PDE)4-selective inhibitor , has recently been approved for treating severe chronic obstructive pulmonary disease ( P48444 ) patients with exacerbation . However , there is also clinical evidence suggesting the development of tachyphylaxis or tolerance on repeated dosing of roflumilast and the possible contribution of Q07343 up-regulation , which could be counterproductive for suppressing inflammation . Thus , understanding how Q07343 is up-regulated in the context of the complex pathogenesis and medications of P48444 may help improve the efficacy and possibly ameliorate the tolerance of roflumilast . Here we show that roflumilast synergizes with nontypeable Haemophilus influenzae ( NTHi ) , a major bacterial cause of P48444 exacerbation , to up-regulate PDE4B2 expression in human airway epithelial cells in vitro and in vivo . Up-regulated PDE4B2 contributes to the induction of certain important chemokines in both enzymatic activity-dependent and activity-independent manners . We also found that protein kinase A catalytic subunit β ( PKA-Cβ ) and nuclear factor-κB ( NF-κB ) p65 subunit were required for the synergistic induction of PDE4B2 . PKA-Cβ phosphorylates p65 in a DB02527 -dependent manner . Moreover , Ser276 of p65 is critical for mediating the PKA-Cβ-induced p65 phosphorylation and the synergistic induction of PDE4B2 . Collectively , our data unveil a previously unidentified mechanism underlying synergistic up-regulation of PDE4B2 via a cross-talk between PKA-Cβ and p65 and may help develop new therapeutic strategies to improve the efficacy of DB05876 inhibitor . DB00501 induces interleukin-18 production through H2-agonist activity in monocytes . The present study demonstrates a possible mechanism for the improvement of gastrointestinal cancer patients ' prognosis by the histamine receptor type 2 ( P25021 ) antagonist cimetidine . This agent , but not the P25021 antagonists ranitidine and famotidine , induced the production of an antitumor cytokine , interleukin ( IL ) -18 , by human monocytes and dendritic cells ( DC ) . In fact , ranitidine and famotidine antagonized cimetidine-induced Q14116 production . DB00501 induced the activation of caspase-1 , which is reported to modify immature Q14116 to mature/active Q14116 , and the elevation of intracellular DB02527 , leading to the activation of protein kinase A ( PKA ) . The PKA inhibitor H89 abolished the Q14116 production induced by cimetidine . Moreover , the effects of cimetidine on Q14116 production were reproduced in peripheral blood mononuclear cells from wild-type mice , but not in those from P25021 knockout mice . In conclusion , cimetidine , a partial agonist for P25021 , has a pharmacological profile different from ranitidine and famotidine , possibly contributing to its antitumor activity on gastrointestinal cancers . Plasmapheresis-resistant acute humoral rejection successfully treated with anti- P01031 antibody . Even if kidney graft survival has improved during the last decades , sensitized pediatric patients are an emerging problem . We describe a 17-yr-old male who lost his first graft due to chronic rejection becoming hyperimmunized ( CDC P06703 99.61 % ) . A desensitization protocol based on high-dose IVIG , PP , and two Mabthera(®) infusions was performed with minor response ( CDC P06703 post-desensitization 80 % ) . One month after his second non-living transplant , he developed a biopsy-proven AMR ; post-transplant immunological monitoring showed the presence of donor-specific anti-DQ5 antibodies ( DSA , MFI 20.000 ) . He received methylprednisolone pulses and 45 PP sessions without clinical response ; eculizumab was then used to salvage a kidney undergoing severe PP-resistant rejection . A biopsy performed after the fourth eculizumab infusion showed complete resolution of AMR . DB01257 infusions were then continued for the first year post-transplantation . Two yr after transplantation , graft function is stable . Anti- P01031 therapy may represent an effective therapeutic option in pediatric patients with PP-resistant AMR . Terminal complement inhibition decreases antibody-mediated rejection in sensitized renal transplant recipients . Sensitized renal transplant recipients with high levels of donor-specific alloantibody ( DSA ) commonly develop antibody-mediated rejection ( AMR ) , which may cause acute graft loss or shorten allograft survival . We examined the efficacy of terminal complement inhibition with the humanized anti- P01031 antibody , eculizumab , in the prevention AMR in renal transplant recipients with a positive crossmatch against their living donor . The incidence of biopsy-proven AMR in the first 3 months posttransplant in 26 highly sensitized recipients of living donor renal transplants who received eculizumab posttransplant was compared to a historical control group of 51 sensitized patients treated with a similar plasma exchange ( PE ) -based protocol without eculizumab . The incidence of AMR was 7.7 % ( 2/26 ) in the eculizumab group compared to 41.2 % ( 21/51 ) in the control group ( p = 0.0031 ) . DB01257 also decreased AMR in patients who developed high levels of DSA early after transplantation that caused proximal complement activation . With eculizumab , AMR episodes were easily treated with PE reducing the need for splenectomy . On 1-year protocol biopsy , transplant glomerulopathy was found to be present in 6.7 % ( 1/15 ) eculizumab-treated recipients and in 35.7 % ( 15/42 ) of control patients ( p = 0.044 ) . Inhibition of terminal complement activation with eculizumab decreases the incidence of early AMR in sensitized renal transplant recipients ( ClincalTrials.gov number NCT006707 ) . Evolution of portal-systemic collateral vasopressin response in endotoxemic portal hypertensive rats . Cirrhotic patients with portal hypertension and variceal hemorrhage are vulnerable to endotoxemia . However , the direct influence of endotoxemia on portal-systemic collateral vasculature remains unexplored . In this study , portal hypertension was induced in Sprague-Dawley rats by partial portal vein ligation . On the 7th day after portal vein ligation , at 0.5 , 1.5 , and 5 h post endotoxin ( LPS ; Escherichia coli serotype O111:B4 , 3 mg/kg , i.p. , E0.5 , E1.5 and O94989 , respectively ) or saline ( control , C0.5 , C1.5 , and P01031 , respectively ) injection , hemodynamic measurements and concentration-response relationships to arginine vasopressin ( AVP ; 10(-10)-10(-7) mol/L ) in collateral vascular bed were obtained . In another six parallel groups , reverse-transcriptase-polymerase chain reaction of P35228 , P29474 , and endothelin 1 ( ET-1 ) mRNA expressions for splenorenal shunt , the most prominent intra-abdominal collateral vessel , was performed . The results showed that E0.5 had lower perfusion pressure changes to AVP and higher splenorenal shunt P29474 expression than C0.5 group ( P < 0.05 ) . Compared with C1.5 , tachycardia , higher perfusion pressure changes and enhanced splenorenal shunt P35228 and ET-1 expression were observed in E1.5 group ( P < 0.05 ) . In O94989 , systemic and portal hypotension with markedly enhanced collateral AVP responsiveness and splenorenal shunt P35228 and ET-1 expressions were noted ( P < 0.05 ) . In conclusion , vasoactive substances counterregulation participates , at least in part , the time-dependent changes of collateral AVP responsiveness in endotoxemic portal hypertensive rats . DB01257 as rescue therapy for atypical hemolytic uremic syndrome with normal platelet count . BACKGROUND : Atypical hemolytic uremic syndrome ( aHUS ) in childhood is a rare disease with frequent progression to end-stage renal disease and a high recurrence after kidney transplantation . DB01257 , a humanized monoclonal antibody that binds to complement protein P01031 , may be beneficial in the treatment of aHUS . CASE-DIAGNOSIS/TREATMENT : A 6-year-old girl developed aHUS with only slightly elevated C3d ( 4.4 % ) , no mutations in complement factors , and no antibodies against factor H . Plasma exchange treatment was successful initially , until aHUS recurred . After reinitiating plasma exchange , normalization of the platelet count and improvement of hemolysis occurred , but renal function worsened . Renal function then improved dramatically promptly after the switch to eculizumab . CONCLUSIONS : This case demonstrates that platelet count is not always a reliable marker for improvement of aHUS and that eculizumab can prevent dialysis in plasma-resistant aHUS patients . DB01257 in an anephric patient with atypical haemolytic uraemic syndrome and advanced vascular lesions . BACKGROUND : Atypical haemolytic uraemic syndrome ( aHUS ) is associated with dysfunction of the alternative pathway of complement . Disease activity subsides as renal failure progresses but recurs upon renal transplantation , indicating that viable renal tissue contributes to disease activity . We present evidence of cerebrovascular occlusive disease indicating that vascular injury may occur in the absence of kidneys . METHODS : A currently 12-year-old girl developed renal failure at the age of 20 months . She underwent bilateral nephrectomy and renal transplantation but lost the transplant due to recurrences . She was on haemodialysis for 7 years . At 10 years of age she developed a transient ischaemic attack . Imaging , genetic investigation and mutation characterization were performed . RESULTS : Imaging demonstrated occlusion and stenosis of the carotid arteries . Two complement mutations , a novel mutation in factor B and a previously described mutation in factor I , and the H3-factor H haplotype , were identified . The factor B mutation , L433S , did not induce excessive complement activation in vitro . Measurement of P01024 degradation products indicated ongoing complement activation . In spite of the patient being anephric , treatment was initiated with eculizumab , a humanized anti- P01031 antibody that blocks terminal complement activation . She underwent a successful kidney transplant 9 months later and has not developed a recurrence or progression of vascular stenosis 1 year later . CONCLUSIONS : The course of disease in this patient with aHUS suggests that complement-mediated vascular injury may occur in the total absence of renal tissue and overt recurrences . To our knowledge , this is the first description of eculizumab treatment in an anephric aHUS patient . Treatment of a patient with classical paroxysmal nocturnal hemoglobinuria and Budd-Chiari syndrome , with complement inhibitor eculizumab : Case Report . Background. Paroxysmal nocturnal haemoglobinuria ( PNH ) is a rare acquired clonal disorder of hematopoietic stem cells involving all blood cells . Erythrocytes have increased susceptibility to complement-mediated haemolysis . Thrombosis is the leading cause of mortality and follows episodes of acute hemolysis . DB01257 , a monoclonal antibody blocking activation of complement P01031 is currently used in the treatment of PNH . Recent results demonstrated that eculizumab effectively reduces thrombosis . Description of case . We present a 30-year-old male patient admitted with abdominal and lumbar pain . Thorough investigation revealed severe hemolytic anemia requiring transfusions and hepatosplenomegaly . Imaging findings were compatible with a Budd-Chiari syndrome . Flow cytometry confirmed the PNH diagnosis . Due to refractory ascites he underwent a transjugular intrahepatic portal-systemic shunt ( TIPS ) and eculizumab administration was started . Results . He has already completed three years of eculizumab treatment and he is transfusion independent . There is also a significant reduction in fatigue with improvement in his quality of life . Doppler scans of his TIPS persistently show it to be patent . Conclusions . Classical PNH patients with thrombosis and severe intravascular hemolysis are particularly challenging to manage . For these patients , eculizumab is a reasonable therapeutic option , expecting that by decreasing the risk for thrombosis , life expectancy may be increased . Learnings from over 25 years of PNH experience : the era of targeted complement inhibition . Paroxysmal nocturnal haemoglobinuria ( PNH ) is a progressive and life-threatening disease that causes thrombosis , end organ damage and impaired quality of life . Chronic uncontrolled complement activation leads to chronic haemolysis , causing progressive morbidities and early mortality . Hence , early diagnosis is essential for improved patient management and prognosis . DB01257 ( SOLIRIS® ) specifically inhibits chronic , uncontrolled complement activation and is the first-in-class , humanised , monoclonal antibody targeting P01031 within the terminal complement pathway . DB01257 is the first and only approved treatment for PNH in adults and children . Backcross and partial advanced intercross analysis of nonobese diabetic gene-mediated effects on collagen-induced arthritis reveals an interactive effect by two major loci . Genetic segregation analysis between NOD and C57BL strains have been used to identify loci associated with autoimmune disease . Only two loci ( Cia2 and Cia9 ) had earlier been found to control development of arthritis , whereas none of the previously identified diabetes loci was of significance for arthritis . We have now made a high-powered analysis of a backcross of NOD genes on to the B10.Q strain for association with collagen-induced arthritis . We could confirm relevance of both Cia2 and Cia9 as well as the interaction between them , but we did not identify any other significant arthritis loci . Immune cellular subtyping revealed that Cia2 was also associated with the number of blood macrophages . Congenic strains of the Cia2 and Cia9 loci on the B10.Q background were made and used to establish a partial advanced intercross ( P05121 ) . Testing the P05121 mice for development of collagen-induced arthritis confirmed the loci and the interactions and also indicated that at least two genes contribute to the Cia9 locus . Furthermore , it clearly showed that Cia2 is dominant protective but that the protection is not complete . Because these results may indicate that the Cia2 effect on arthritis is not only due to the deficiency of the complement P01031 , we analyzed complement functions in the Cia2 congenics as well as the P05121 mice . These data show that not only arthritis but also P01031 -dependent complement activity is dominantly suppressed , confirming that P01031 is one of the major genes explaining the Cia2 effect . Blockade of alternative complement pathway in dense deposit disease . A patient aged 17 with dense deposit disease associated with complement activation , circulating P01024 Nef , and Factor H mutation presented with nephrotic syndrome and hypertension . Steroid therapy , plasma exchange , and rituximab failed to improve proteinuria and hypertension despite a normalization of the circulating sC5b9 complex . DB01257 , a monoclonal antibody directed against P01031 , was used to block the terminal product of the complement cascade . The dose was adapted to achieve a CH50 below 10 % , but proteinuria and blood pressure were not improved after 3 months of treatment . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . DB01257 reduces complement activation , inflammation , endothelial damage , thrombosis , and renal injury markers in aHUS . Atypical hemolytic uremic syndrome ( aHUS ) is a genetic , life-threatening disease characterized by uncontrolled complement activation , systemic thrombotic microangiopathy ( TMA ) , and vital organ damage . We evaluated the effect of terminal complement blockade with the anti- P01031 monoclonal antibody eculizumab on biomarkers of cellular processes involved in TMA in patients with aHUS longitudinally , during up to 1 year of treatment , compared with in healthy volunteers . Biomarker levels were elevated at baseline in most patients , regardless of mutational status , plasma exchange/infusion use , platelet count , or lactate dehydrogenase or haptoglobin levels . DB01257 reduced terminal complement activation ( C5a and sC5b-9 ) and renal injury markers ( clusterin , cystatin-C , β2-microglobulin , and liver fatty acid binding protein-1 ) to healthy volunteer levels and reduced inflammation ( soluble tumor necrosis factor receptor-1 ) , coagulation ( prothrombin fragment F1+2 and d-dimer ) , and endothelial damage ( thrombomodulin ) markers to near-normal levels . Alternative pathway activation ( Ba ) and endothelial activation markers ( soluble vascular cell adhesion molecule-1 ) decreased but remained elevated , reflecting ongoing complement activation in aHUS despite complete terminal complement blockade . These results highlight links between terminal complement activation and inflammation , endothelial damage , thrombosis , and renal injury and underscore ongoing risk for systemic TMA and progression to organ damage . Further research regarding underlying complement dysregulation is warranted . This trial was registered at www.clinicaltrials.gov as # NCT01194973 . DB01257 for dense deposit disease and P01024 glomerulonephritis . BACKGROUND AND OBJECTIVES : The principle defect in dense deposit disease and P01024 glomerulonephritis is hyperactivity of the alternative complement pathway . DB01257 , a monoclonal antibody that binds to P01031 to prevent formation of the membrane attack complex , may prove beneficial . DESIGN , SETTING , PARTICIPANTS , & MEASUREMENTS : In this open-label , proof of concept efficacy and safety study , six subjects with dense deposit disease or P01024 glomerulonephritis were treated with eculizumab every other week for 1 year . All had proteinuria > 1 g/d and/or AKI at enrollment . Subjects underwent biopsy before enrollment and repeat biopsy at the 1-year mark . RESULTS : The subjects included three patients with dense deposit disease ( including one patient with recurrent dense deposit disease in allograft ) and three patients with P01024 glomerulonephritis ( including two patients with recurrent P01024 glomerulonephritis in allograft ) . Genetic and complement function testing revealed a mutation in P08603 and MCP in one subject each , P01024 nephritic factor in three subjects , and elevated levels of serum membrane attack complex in three subjects . After 12 months , two subjects showed significantly reduced serum creatinine , one subject achieved marked reduction in proteinuria , and one subject had stable laboratory parameters but histopathologic improvements . Elevated serum membrane attack complex levels normalized on therapy and paralleled improvements in creatinine and proteinuria . CONCLUSIONS : Clinical and histopathologic data suggest a response to eculizumab in some but not all subjects with dense deposit disease and P01024 glomerulonephritis . Elevation of serum membrane attack complex before treatment may predict response . Additional research is needed to define the subgroup of dense deposit disease/ P01024 glomerulonephritis patients in whom eculizumab therapy can be considered . [ Anti-cholesterol agents , new therapeutic approaches ] . Statins and fibrates constitute the two major families of lipid-lowering agents . Statins are widely used for the treatment of pure hypercholesterolaemia while fibrates are used for the treatment of hypertriglyceridemia . Both drugs are also used for the treatment of mixed dyslipidemia . Some fibrates efficiently lower serum LDL-cholesterol . Statins inhibit P04035 and decrease cellular cholesterol synthesis . The resulting lower intracellular cholesterol concentration induces the activation of SREBP thus inducing the over expression and transcription of the P01130 gene . This over expression of the P01130 in the liver increases the clearance of circulating LDL thus decreasing the LDL-cholesterol plasma levels . The effects of fibrates on lipid metabolism are entirely due to their capacity to activate Q07869 and to induce the over expression of genes containing a PPRE in their promoter . Fibrates decrease triglyceride concentrations by increasing the beta-oxidation of fatty acids in the liver and by decreasing triglyceride-VLDL synthesis . Fibrates also decrease triglycerides by increasing the hydolysys of triglycerides in chylomicron and VLDL through their capacity to increase and to decrease the lipoprotein lipase and the apo C-III transcription , respectively . Fibrates could decrease triglycerides partly by inducing apo A-V over-expression . These molecules increase HDL-cholesterol by increasing apo A-I and apo A-II transcription . Therefore the mechanisms of action of statins and fibrates depend on their capacity to modulate the expression of genes controlling lipoprotein metabolism .

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Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . [ Clinical effect and side effect of troglitazone ] . DB00197 , a P37231 agonist , is a new drug for type 2 diabetes . The drug decreases blood glucose via enhancing insulin action . Recently Sankyo pharmaceutical company is warning severe hepatotoxicity by troglitazone . It recommends to examine liver function every month in diabetic patients treated with the drug in order early to find drug-induced hepatitis . In Japan 153 diabetic patients treated with the drug developed severe hepatitis and 8 of them died of drug-side effects . Quinone metabolite of troglitazone predominantly in the liver to a sulfate conjugate and activation of Q07869 gamma and O75469 ( pregnane X receptor ) by troglitazone are supposed to be factors of hepatotoxic mechanism . Recombinant growth hormone : a new cardiovascular drug therapy . GH has an important role in normal cardiovascular physiologic functioning , working indirectly through effects on DB01277 . An excess or deficiency of GH causes an increased rate of cardiovascular disease , including cardiomyopathy . A relative GH deficiency in older subjects may also increase cardiovascular morbidity and mortality risk . In replacement doses , GH can enhance myocardial contractility ; can decrease peripheral vascular resistance ; and can reduce total cholesterol and LDL-cholesterol values and fibrinogen and P05121 levels . These effects of GH , coupled with the ability to improve skeletal muscle function and reduce adiposity , make it an attractive treatment for patients with CHF and a potential maintenance drug for elderly people . Clinical trials , including studies with P01286 that may reduce the adverse effects of GH therapy , such as hyperglycemia and hypertension , are now in progress . Thiazolidinediones downregulate stearoyl- DB01992 desaturase 1 gene expression in 3T3- Q9NUQ9 adipocytes . Thiazolidinediones ( TZDs ) are known to have potent increases of insulin sensitivity . Because peroxisome proliferator-activated receptor-gamma ( P37231 ) , a receptor for TZDs , is mainly expressed in adipocytes , we tried to search the TZD-targeted genes in mouse 3T3- Q9NUQ9 adipocytes . By the mRNA differential display method , one band repressed by troglitazone was obtained , which corresponded to the partial sequences of the stearoyl- DB01992 desaturase 1 ( SCD1 ) gene . DB00197 dramatically decreased SCD1 mRNA levels in 3T3- Q9NUQ9 adipocytes in a dose-dependent manner . Pioglitazone also repressed the SCD1 mRNA expression , whereas WY-14,643 had no apparent effect . Both troglitazone and pioglitazone raised the composition ( weight percentage ) of myristic acid ( C14:0 ) , palmitic acid ( C16:0 ) , and stearic acid ( C18:0 ) , but lowered the composition of the delta9-cis desaturated fatty acids such as myristoleic acid ( C14:1 , delta9 ) , palmitoleic acid ( C16:1 , delta9 ) , oleic acid ( C18:1 , delta9 ) , and linoleic acid ( C18:2 , delta9,12 ) . These results indicate that TZDs repress SCD1 activity in 3T3- Q9NUQ9 adipocytes via downregulating SCD1 enzyme gene expression . Delineating biological pathways unique to embryonic stem cell-derived insulin-producing cell lines from their noninsulin-producing progenitor cell lines . To identify unique biochemical pathways in embryonic stem cell-derived insulin-producing cells as potential therapeutic targets to prevent or delay beta-cell dysfunction or death in diabetic patients , comparative genome-wide gene expression studies of recently derived mouse insulin-producing cell lines and their progenitor cell lines were performed using microarray technology . Differentially expressed genes were functionally clustered to identify important biochemical pathways in these insulin-producing cell lines . Biochemical or cellular assays were then performed to assess the relevance of these pathways to the biology of these cells . A total of 185 genes were highly expressed in the insulin-producing cell lines , and computational analysis predicted the pentose phosphate pathway ( PPP ) , clathrin-mediated endocytosis , and the peroxisome proliferator-activated receptor ( Q07869 ) signaling pathway as important pathways in these cell lines . P01308 -producing ERoSHK cells were more resistant to hydrogen peroxide ( H(2)O(2) ) -induced oxidative stress . Inhibition of PPP by dehydroepiandrosterone and 6-aminonicotinamide abrogated this H(2)O(2) resistance with a concomitant decrease in PPP activity as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ( MTT ) assay . Clathrin-mediated endocytosis , which is essential in maintaining membrane homeostasis in secreting cells , was up-regulated by glucose in ERoSHK but not in their progenitor ERoSH cells . Its inhibition by chlorpromazine at high glucose concentration was toxic to the cells . DB00197 , a P37231 agonist , up-regulated expression of Ins1 and Ins2 but not Glut2 . Gene expression analysis has identified the PPP , clathrin-mediated endocytosis , and the Q07869 signaling pathway as the major delineating pathways in these insulin-producing cell lines , and their biological relevance was confirmed by biochemical and cellular assays . Differential regulation by troglitazone of plasminogen activator inhibitor type 1 in human hepatic and vascular cells . DB00197 , a novel oral insulin sensitizer , normalizes increased plasma activity of plasminogen activator inhibitor type 1 ( P05121 ) in hyperinsulinemic patients such as women with polycystic ovary syndrome and patients with type 2 diabetes mellitus . However , underlying mechanisms have not yet been fully elucidated . Human hepatic and vascular cells , the main sources of circulating P05121 , were studied in cell culture . In human hepatic cells , P05121 accumulated in conditioned medium by 23 % within 24 h after exposure to 3 microg/mL troglitazone ( P = 0.001 ) . The accumulation depended on the concentration of troglitazone , but not that of insulin ( known to stimulate P05121 synthesis ) . By contrast , in human aortic smooth muscle cells , 3 microg/mL troglitazone decreased basal P05121 expression by 23 % ( P = 0.037 ) and decreased transforming growth factor-beta-induced expression by 34 % ( P = 0.026 ) . Concomitant insulin had no effect . P00750 was decreased by 38 % ( P = 0.002 ) . In human endothelial cells , P05121 was diminished by 32 % ( P < 0.001 ) , whereas tissue-type plasminogen activator was unaffected . The results suggest that the reduction in plasma activity of P05121 induced by troglitazone in patients may reflect both directly mediated diminution of its elaboration from vessel walls and indirectly mediated reduction of its hepatic synthesis secondary to attenuation of hyperinsulinemia ( known to increase the hepatic synthesis of P05121 ) . Effect of troglitazone on P04798 induction . Several peroxisome proliferators enhance P04798 activity , but the mechanisms involved in this enhancement remain unknown . In this study , we examined the effect of troglitazone , a peroxisome proliferator-activated receptor-gamma ( P37231 ) agonist , on P04798 gene expression and explored the mechanisms involved in these effects . DB00197 increased gene expression of P04798 mRNA and also increased P04798 -specific 7-ethoxyresorufin-O-deethylase ( EROD ) activity in a dose-dependent manner . Moreover , concomitant treatment with troglitazone and GW9662 , a Q07869 antagonist , markedly reduced the troglitazone-inducible EROD activity . Luciferase reporter assays using Hepa-1c1c7 cells showed a significant transactivation by troglitazone with a reporter plasmid containing a region from -1395 to +7 of the P04798 gene . We found that a putative peroxisome proliferator-response element ( PPRE ) between -521 and -500 is located in the P04798 gene promoter . Their inactivation by deletion mutagenesis suppressed the inductive effect of troglitazone on P04798 promoter activation . Electrophoretic mobility shift assay revealed that troglitazone induced the activation of the P37231 to a form capable of binding specifically to the PPRE sequence of the P04798 gene promoter . Furthermore , troglitazone increased the formation of the benzo[a]pyrene ( BaP ) -DNA adduct . Overall , our results suggest that troglitazone induces P04798 enzyme activity and gene expression through P37231 activation , and may be involved in carcinogenesis . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . Anti-inflammatory effects of troglitazone in nondiabetic obese subjects independent of changes in insulin sensitivity . BACKGROUND : Obesity is characterised by insulin resistance and by elevated levels of proinflammatory markers . We investigated whether , in the absence of changes in glucose , thiazolidinediones ( TZDs ) have anti-inflammatory effects and whether improvement of insulin sensitivity correlates with suppression of inflammatory markers . METHODS : We performed a randomised double-blind placebo-controlled crossover study with troglitazone ( 400 mg daily for eight weeks ) in 15 normoglycaemic obese subjects . We measured plasma high-sensitivity P02741 ( hsCRP ) , interleukin-6 ( P05231 ) , leptin , tissue-type plasminogen activator ( tPA ) , plasminogen activator inhibitor-1 ( P05121 ) and tumour necrosis factor-alpha ( P01375 ) after each of the two treatment periods and in 13 age- and sex-matched lean individuals . RESULTS : Obese subjects were insulin resistant ( decreased glucose infusion rate ( GIR ) during euglycaemic hyperinsulinaemic clamp ) and had higher plasma levels of hsCRP , P05231 , leptin , tPA , and P05121 compared with lean subjects . P01375 also tended to be higher . DB00197 improved insulin sensitivity ( mean increase in whole body glucose uptake 23.1 +/- 10.5 % ( p = 0.047 ) ) and normalised plasma concentrations of hsCRP , tPA and P01375 , whereas it did not significantly change P05231 , leptin and P05121 . Changes in GIR did not correlate with changes in inflammatory markers . CONCLUSION : DB00197 induces suppression of some of the inflammatory markers that are elevated in normoglycaemic obese subjects . The suppression of inflammatory markers , however , does not correlate with improvement in insulin sensitivity , suggesting involvement of partially differential mechanisms in these effects of TZDs . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Synthetic peroxisome proliferator-activated receptor-gamma agonists restore impaired vasorelaxation via DB00171 -sensitive K+ channels by high glucose . The present study was designed to examine whether in the human artery , synthetic peroxisome proliferator-activated receptor ( Q07869 ) -gamma agonists restore vasorelaxation as well as hyperpolarization via DB00171 -sensitive K+ channels impaired by the high concentration of D-glucose and whether the restoration may be mediated by the antioxidant capacity of these agents . The isometric force and membrane potential of human omental arteries without endothelium were recorded . The production rate of superoxide was evaluated using a superoxide-generating system with xanthine-xanthine oxidase in the absence of smooth muscle cells . DB01016 abolished vasorelaxation and hyperpolarization in response to levcromakalim . Addition of D-glucose ( 20 mM ) but not L-glucose ( 20 mM ) reduced this vasorelaxation and hyperpolarization . Synthetic P37231 agonists ( troglitazone and rosiglitazone ) and/or an inhibitor of superoxide generation ( 4,5-dihydroxy-1,3-benzene-disulfonic acid , Tiron ) , but not a Q07869 agonist ( fenofibrate ) , restored vasorelaxation and hyperpolarization in response to levcromakalim in arteries treated with D-glucose . DB00197 and rosiglitazone , but not fenofibrate , decreased the production rate of superoxide without affecting uric acid generation . These findings suggest that synthetic P37231 agonists recover the function of DB00171 -sensitive K+ channels reduced by the high concentration of glucose in human vascular smooth muscle cells and that the effect of these agonists may be mediated in part by their antioxidant capacity . [ Dyslipidemia in insulin resistance and its improvement by troglitazone ] . Dyslipideia in insulin resistant state are characterized by 3 major components ; increased triglyceride levels , decreased high-density lipoprotein ( HDL ) cholesterol , and change in the composition of low-density lipoprotein ( LDL ) cholesterol particle which results in small-dense LDL . P01308 resistance is thought to lead to overproduction of very low-density lipoprotein ( VLDL ) cholesterol through the decreased peripheral lipoprotein lipase ( P06858 ) activity , increased production of apolipoprotein B-100 and decreased clearance of remnant particles . DB00197 , an insulin action enhancer , decreases the triglyceride level , increases HDL cholesterol level and decreases th proportion of small-dense LDL through the direct effect on lipoprotein metabolism . However , activation of P37231 is considered to possese potential atherogeneity and more closer examination is needed . DB04946 binding to human and rat dopamine and 5-HT receptors . DB04946 ( DB04946 ; 1- [ 4-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] -3- methoxyphenyl ] ethanone ) is a compound currently in clinical trials for the treatment of schizophrenia . DB04946 displays affinity for dopamine D2 receptors and for 5- Q13049 receptors and has a variety of in vivo activities suggestive of an atypical antipsychotic . Here we present an examination of the affinity of iloperidone to a variety of human and rat homologs of dopamine and 5-HT receptor subtypes . We employed receptor binding assays using membranes from cells stably expressing human dopamine D1 , D2S , D2L , D3 , D4 and D5 and 5- Q13049 and P28335 receptors and rat P50406 and P34969 receptors . DB04946 displayed higher affinity for the dopamine D3 receptor ( Ki = 7.1 nM ) than for the dopamine D4 receptor ( Ki = 25 nM ) . DB04946 displayed high affinity for the P50406 and P34969 receptors ( Ki = 42.7 and 21.6 nM , respectively ) , and was found to have higher affinity for the 5- Q13049 ( Ki = 5.6 nM ) than for the P28335 receptor ( Ki = 42.8 nM ) . The potential implications of this receptor binding profile are discussed in comparison with data for other antipsychotic compounds . 2(3H)-benzoxazolone and bioisosters as " privileged scaffold " in the design of pharmacological probes . The 2(3H)-benzoxazolone heterocycle and its bioisosteric surrogates ( such as 2(3H)-benzothiazolinone , benzoxazinone , etc. ) have received considerable attention from the medicinal chemists owing to their capacity to mimic a phenol or a catechol moiety in a metabolically stable template . These heterocycles and pyrocatechol have indeed similar pKa 's , electronic charge distribution , and chemical reactivity . Therapeutic applications of this template are very broad , and range from analgesic anti-inflammatory compounds ( including P37231 antagonists ) to antipsychotic and neuroprotective anticonvulsant compounds . High affinity ligands have been obtained also for dopaminergic ( D2 and D4 ) , serotoninergic ( P08908 and P28223 ) , sigma-1 and sigma-2 receptors . Owing to the high number of positive hits encountered with this heterocycle and its congeners , 2(3H)-benzoxazolone template certainly deserves the title of " privileged scaffold " in medicinal chemistry . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . The role of serotonin receptor subtypes in the behavioural effects of neuroleptic drugs . A paw test study in rats . The present study was designed to evaluate the roles of serotonin P08908 and 5-HT2 receptors in the effects of neuroleptic drugs in the paw test . This behavioural test has been shown to model both the antipsychotic efficacy as well as the extrapyramidal side-effect liability of neuroleptic drugs . Whereas the P08908 receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OHDPAT ) reduced the effects of the classical neuroleptic haloperidol , it increased the effects of the atypical neuroleptic clozapine . The 5-HT2 receptor antagonist ketanserin as well as the P28335 /5-HT2 receptor antagonist ritanserin , on the other hand reduced the effects of haloperidol , whereas the P28335 /5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane ( DOI ) reduced the effects of clozapine . The most important finding , however , was that the behavioural effects of different ( putative ) neuroleptics ( fluphenazine , P35240 -39166 , remoxipride , prothipendyl , thioridazine and risperidone ) were differentially influenced by both 8-OHDPAT and DOI , suggesting that there are important differences between the neuronal mechanisms underlying the behavioural effects of these neuroleptic drugs , even within the subclasses of classical and atypical neuroleptics . Neuronal ablation of p-Akt at Ser473 leads to altered P08908 /2A receptor function . The serotonergic system regulates a wide range of behavior , including mood and impulsivity , and its dysregulation has been associated with mood disorders , autism spectrum disorder , and addiction . Diabetes is a risk factor for these conditions . P01308 resistance in the brain is specifically associated with susceptibility to psychostimulant abuse . Here , we examined whether phosphorylation of Akt , a key regulator of the insulin signaling pathway , controls serotonin ( 5-HT ) signaling . To explore how impairment in Akt function regulates 5-HT homeostasis , we used a brain-specific rictor knockout ( KO ) mouse model of impaired neuronal phosphorylation of Akt at Ser473 . Cortical P08908 and 5- Q13049 receptor binding was significantly elevated in rictor KO mice . Concomitant with this elevated receptor expression , the P08908 receptor agonist 8-Hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT ) led to an increased hypothermic response in rictor KO mice . The increased cortical P08908 receptor density was associated with higher P08908 receptor levels on the cortical cell surface . In contrast , rictor KO mice displayed significantly reduced head-twitch response ( HTR ) to the 5- Q13049 /C agonist 2,5-dimethoxy-4-iodoamphetamine ( DOI ) , with evidence of impaired 5- Q13049 /C receptor signaling . In vitro , pharmacological inhibition of Akt significantly increased P08908 receptor expression and attenuated DOI-induced 5- Q13049 receptor signaling , thereby lending credence to the observed in vivo cross-talk between neuronal Akt signaling and 5-HT receptor regulation . These data reveal that defective central Akt function alters 5-HT signaling as well as 5-HT-associated behaviors , demonstrating a novel role for Akt in maintaining neuronal 5-HT receptor function . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . P37231 reduces the growth rate of pancreatic cancer cells through the reduction of cyclin D1 . P37231 ( PPARgamma ) forms a heterodimeric DNA-binding complex with the retinoid X receptor ( RXR ) and regulates the transcription of its target genes . Activation of PPARgamma has been shown to induce P55008 arrest and to inhibit cell growth of human pancreatic carcinoma cell lines . The purpose of the present study was to examine the effect of ligand activation of PPARgamma and RXR on cell growth and on the expression of P55008 cyclins in a pancreatic cancer cell line PANC-1 , which expresses PPARgamma at high levels . DB00197 , a specific ligand for PPARgamma , was found to cause a reduction in the growth rate and induced P55008 cell cycle arrest and this effect was additive with that of 9-cis retinoic acid ( 9-cis RA ) , a ligand for RXR . Of the P55008 cyclins tested , troglitazone specifically reduced the expression of cyclin D1 mRNA and the corresponding protein and this effect was also additive with 9-cis RA . These results suggest that the activation of PPARgamma together with RXR may be useful for the suppression of pancreatic cancer cell growth through the reduction in cyclin D1 levels . DB00197 enhances tamoxifen-induced growth inhibitory activity of MCF-7 cells . P37231 ( PPARgamma ) ligands have been identified as a potential source of therapy for human cancers . However , PPARgamma ligands have a limitation for breast cancer therapy , since estrogen receptor alpha ( ER(alpha) ) negatively interferes with PPARgamma signaling in breast cancer cells . Here we show that ER(alpha) inhihits PPARgamma transactivity and ER(alpha)-mediated inhibition of PPARgamma transactivity is blocked by tamoxifen , an estrogen receptor blocker . The activation of ER(alpha) with 17-beta-estradiol blocked PPRE transactivity induced by troglitazone , a PPARgamma ligand , indicating the resistance of ER(alpha)-positive breast cancer cells to troglitazone . Indeed , troglitazone inhibited the growth of ER(alpha)-negative MDA-MB-231 cells more than that of ER(alpha)-positive MCF-7 cells . Combination of troglitazone with tamoxifen led to a marked increase in growth inhibition of ER(alpha)-positive MCF-7 cells compared to either agent alone . Our data indicates that troglitazone enhances the growth inhibitory activity of tamoxifen in ER(alpha)-positive MCF-7 cells . Androgens stimulate myogenic differentiation and inhibit adipogenesis in C3H 10T1/2 pluripotent cells through an androgen receptor-mediated pathway . DB00624 supplementation increases skeletal muscle mass and decreases fat mass ; however , the underlying mechanisms are unknown . We hypothesized that testosterone regulates body composition by promoting the commitment of mesenchymal pluripotent cells into myogenic lineage and inhibiting their differentiation into adipogenic lineage . Mouse C3H 10T1/2 pluripotent cells were treated with testosterone ( 0-300 nM ) or dihydrotestosterone ( DB02901 , 0-30 nM ) for 0-14 d , and myogenic conversion was evaluated by immunocytochemical staining for early ( MyoD ) and late ( myosin heavy chain II ; MHC ) myogenic markers and by measurements of MyoD and MHC mRNA and protein . Adipogenic differentiation was assessed by adipocyte counting and by measurements of peroxisomal proliferator-activated receptor gamma 2 ( Q07869 gamma 2 ) mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . The number of MyoD+ myogenic cells and MHC+ myotubes and MyoD and MHC mRNA and protein levels increased dose dependently in response to testosterone and DB02901 treatment . Both testosterone and DB02901 decreased the number of adipocytes and down-regulated the expression of Q07869 gamma 2 mRNA and Q07869 gamma 2 protein and CCAAT/enhancer binding protein alpha . P10275 mRNA and protein levels were low at baseline but increased after testosterone or DB02901 treatment . The effects of testosterone and DB02901 on myogenesis and adipogenesis were blocked by bicalutamide . Therefore , testosterone and DB02901 regulate lineage determination in mesenchymal pluripotent cells by promoting their commitment to the myogenic lineage and inhibiting their differentiation into the adipogenic lineage through an androgen receptor-mediated pathway . The observation that differentiation of pluripotent cells is androgen dependent provides a unifying explanation for the reciprocal effects of androgens on muscle and fat mass in men . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Effects of peroxisome proliferator-activated receptor-alpha and -gamma agonist , DB04971 , on diabetic complications in Zucker diabetic fatty rats . This study has investigated the effects of DB04971 , a peroxisome proliferator-activated receptor ( Q07869 ) -alpha and P37231 agonist , on the pathogenesis of diabetic complications in the Zucker diabetic fatty ( ZDF ) rats , a model of type 2 diabetes . Comparison is made with troglitazone , a P37231 agonist . The ZDF rats exhibited hyperglycaemia and hyperlipidaemia , and developed diabetic complications such as cataract , nephropathy , and neuropathy . Treatment with DB04971 from the prediabetic stage controlled glycaemia and lipidaemia , and prevented the development of diabetic complications . DB00197 was less effective in controlling serum cholesterol and neuropathy . ZDF rats developed diabetic osteopenia with reduced bone turnover , and this was prevented by DB04971 and troglitazone , possibly mediated by increased bone turnover and bone formation . Since DB04971 controlled glycaemia and lipidaemia in ZDF rats and prevented several diabetic complications , it is suggested that treatment with DB04971 , which activates both Q07869 and P37231 , could provide a valuable therapeutic approach against diabetic complications in type 2 diabetes . Reduced P25874 -1 content in in vitro differentiated beige/brite adipocytes derived from preadipocytes of human subcutaneous white adipose tissues in obesity . INTRODUCTION : Brown adipose tissue ( Q14032 ) is a potential therapeutic target to reverse obesity . The purpose of this study was to determine whether primary precursor cells isolated from human adult subcutaneous white adipose tissue ( WAT ) can be induced to differentiate in-vitro into adipocytes that express key markers of brown or beige adipose , and whether the expression level of such markers differs between lean and obese young adult males . METHODS : Adipogenic precursor cells were isolated from lean and obese individuals from subcutaneous abdominal WAT biopsies . Cells were grown to confluence , differentiated for 2.5 weeks then harvested for measurement of gene expression and P25874 protein . RESULTS : There was no difference between groups with respect to differentiation into adipocytes , as indicated by oil red-O staining , rates of lipolysis , and expression of adipogenic genes ( P15090 , P37231 ) . WAT genes ( P31274 , P06400 ) were expressed equally in the two groups . Post differentiation , the beige adipose specific genes Q99966 and Q07011 were significantly increased in both groups , but classic Q14032 markers Q15915 and Q68G74 decreased significantly . Cell lines from both groups also equally increased post-differentiation expression of the thermogenic-responsive gene Q9UBK2 ( P20142 -1α ) . P25874 gene expression was undetectable prior to differentiation , however after differentiation both gene expression and protein content were increased in both groups and were significantly greater in cultures from lean compared with obese individuals ( p < 0.05 ) . CONCLUSION : Human subcutaneous WAT cells can be induced to attain Q14032 characteristics , but this capacity is reduced in WAT cells from obese individuals . P01308 signaling inhibits the P28335 receptor in choroid plexus via Q96HU1 kinase . BACKGROUND : G protein-coupled receptors ( GPCRs ) interact with heterotrimeric GTP-binding proteins ( G proteins ) to modulate acute changes in intracellular messenger levels and ion channel activity . In contrast , long-term changes in cellular growth , proliferation and differentiation are often mediated by tyrosine kinase receptors and certain GPCRs by activation of mitogen-activated protein ( Q96HU1 ) kinases . Complex interactions occur between these signaling pathways , but the specific mechanisms of such regulatory events are not well-understood . In particular it is not clear whether GPCRs are modulated by tyrosine kinase receptor- Q96HU1 kinase pathways . RESULTS : Here we describe tyrosine kinase receptor regulation of a GPCR via Q96HU1 kinase . P01308 reduced the activity of the P28335 receptor in choroid plexus cells which was blocked by the Q96HU1 kinase kinase ( MEK ) inhibitor , PD 098059 . We demonstrate that the inhibitory effect of insulin and insulin-like growth factor type 1 ( DB01277 ) on the P28335 receptor is dependent on tyrosine kinase , DB01367 and Q96HU1 kinase . The effect may be receptor-specific : insulin had no effect on another GPCR that shares the same G protein signaling pathway as the P28335 receptor . This effect is also direct : activated Q96HU1 kinase mimicked the effect of insulin , and removing a putative Q96HU1 kinase site from the P28335 receptor abolished the effect of insulin . CONCLUSION : These results show that insulin signaling can inhibit P28335 receptor activity and suggest that Q96HU1 kinase may play a direct role in regulating the function of a specific GPCR . P37231 ligands modulate effects of LPS in stimulated rat synovial fibroblasts . This work demonstrated the constitutive expression of peroxisome proliferator-activated receptor ( Q07869 ) -gamma and Q07869 in rat synovial fibroblasts at both mRNA and protein levels . A decrease in P37231 expression induced by 10 microg/ml lipopolysaccharide ( LPS ) was observed , whereas Q07869 mRNA expression was not modified . 15-Deoxy-Delta(12,14)-prostaglandin J(2) ( 15d-PGJ(2) ) dose-dependently decreased LPS-induced cyclooxygenase ( P36551 ) -2 ( -80 % ) and inducible nitric oxide synthase ( P35228 ) mRNA expression ( -80 % ) , whereas troglitazone ( 10 microM ) only inhibited P35228 mRNA expression ( -50 % ) . 15d-PGJ(2) decreased LPS-induced interleukin ( IL ) -1 beta ( -25 % ) and tumor necrosis factor ( P01375 ) -alpha ( -40 % ) expression . Interestingly , troglitazone strongly decreased P01375 expression ( -50 % ) but had no significant effect on P01584 expression . 15d-PGJ(2) was able to inhibit DNA-binding activity of both nuclear factor ( NF ) -kappa B and AP-1 . DB00197 had no effect on NF-kappa B activation and was shown to increase LPS-induced AP-1 activation . 15d-PGJ(2) and troglitazone modulated the expression of LPS-induced P35228 , P35354 , and proinflammatory cytokines differently . Indeed , troglitazone seems to specifically target P01375 and P35228 pathways . These results offer new insights in regard to the anti-inflammatory potential of the P37231 ligands and underline different mechanisms of action of 15d-PGJ(2) and troglitazone in synovial fibroblasts . Effects of C-phycocyanin and Spirulina on salicylate-induced tinnitus , expression of DB01221 receptor and inflammatory genes . Effects of C-phycocyanin ( C-PC ) , the active component of Spirulina platensis water extract on the expressions of N-methyl D-aspartate receptor subunit 2B ( Q13224 ) , tumor necrosis factor-α ( P01375 -α ) , interleukin-1β ( IL-1β ) , and cyclooxygenase type 2 ( P35354 ) genes in the cochlea and inferior colliculus ( IC ) of mice were evaluated after tinnitus was induced by intraperitoneal injection of salicylate . The results showed that 4-day salicylate treatment ( unlike 4-day saline treatment ) caused a significant increase in Q13224 , P01375 -α , and IL-1β mRNAs expression in the cochlea and IC . On the other hand , dietary supplementation with C-PC or Spirulina platensis water extract significantly reduced the salicylate-induced tinnitus and down-regulated the mRNAs expression of Q13224 , P01375 -α , IL-1β mRNAs , and P35354 genes in the cochlea and IC of mice . The changes of protein expression levels were generally correlated with those of mRNAs expression levels in the IC for above genes . Further insight on the hypoglycemic and nonhypoglycemic effects of troglitazone 400 or 600 mg/d : effects on the very-low-density and high-density lipoprotein particle distribution . The effects of troglitazone 400 or 600 mg/d on the glycemic control , very-low-density lipoprotein ( VLDL ) , and high-density lipoprotein ( HDL ) subclass concentrations and plasminogen-activator inhibitor 1 ( P05121 ) levels were assessed in patients with type 2 diabetes that had not been controlled with dietary treatment . This was a multicenter , open-label , parallel-groups study . It included a run-in 4-week diet period and a 24-week randomized treatment . Fifty one patients received 400 mg/d and 55 patients 600 mg . The mean HbA(1c) concentration at the end of the study was similar for both doses . DB00197 , regardless of dose , significantly improved insulin sensitivity assessed by the homeostasis model ( HOMA ) . P05121 levels were significantly decreased in both groups by 13 % . Higher HDL cholesterol concentrations and lower triglycerides levels were observed at the end of treatment . Triglyceride contents were reduced only in the lighter VLDL1 . The change in HDL cholesterol concentration resulted from a combination of increased HDL3 cholesterol and lower HDL2 cholesterol levels . No differences were found in the effects of both treatment groups on the evaluated parameters . Our data provide new information about the actions of the drug on the lipid profile . DB00197 reduces triglyceride levels by lowering the triglycerides content of the VLDL1 particles and increases HDL cholesterol concentrations by increasing HDL3 cholesterol levels . P28335 receptor involvement in female rat lordosis behavior . Adult , hormone-primed , ovariectomized rats ( P05231 -344 ) with bilateral implants within the ventromedial nucleus of the hypothalamus ( VMN ) , were injected with 0.5 microgram estradiol benzoate followed 48 h later with 500 microgram progesterone . This priming produced rats with 2 different levels of sexual receptivity . Rats with a lordosis to mount ratio ( L/M ) >/= 0.5 were used to examine the potential lordosis-inhibiting effects of the 5- Q13049 receptor antagonist , R(+)-a- ( 2 , 3-dimethoxyphenyl ) -1-[2(4-fluoro-phenylethyl)]-4-piperidine-methanol ( MDL 100,907 ) , and the P28335 receptor antagonist , 5-methyl-1-(3-pyridylcarbamoyl)-1,2,3,5-tetrahydropyrrolo [ 2 , 3-f ] indole ( SB 206553 ) . Rats with low sexual receptivity ( L/M < 0.5 ) were bilaterally infused with the 5- Q13049 /2C receptor agonist , (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl ( DOI ) , or DOI plus either MDL 100,907 or SB 206553 to determine if either drug would attenuate the lordosis-facilitating effects of DOI . The P28335 receptor antagonist , but not the 5- Q13049 receptor antagonist , effectively inhibited lordosis behavior . Similarly , SB 206553 was more effective than MDL 100,907 in reducing the DOI-induced increase in lordosis responding . However , both drugs limited the duration of lordosis responding initiated by DOI . These results are consistent with prior suggestions that 5- Q13049 /2C receptors within the VMN are involved in the modulation of lordosis behavior and lead to the suggestion that P28335 , rather than 5- Q13049 , receptors are primarily responsible for the effects of 5-HT2 receptor-active drugs on lordosis behavior . Mediator subunit Gal11p/ Q96RN5 is required for fatty acid-dependent gene activation by yeast transcription factor Oaf1p . The yeast zinc cluster transcription factor Oaf1p activates transcription of target genes in response to direct binding of fatty acids in a manner analogous to the vertebrate nuclear receptor peroxisome proliferator-activated receptoralpha ( PPARalpha ) . PPARs and other metazoan nuclear receptors productively engage several distinct LXXLL motif-containing co-activators , including P52701 family members and the Q15648 /MED1 subunit of the Mediator co-activator , to promote ligand-dependent gene activation . Yeast , however , does not appear to harbor LXXLL motif co-activators , and the mechanism of fatty acid-dependent gene activation by the yeast PPARalpha analog Oaf1p is unknown . Here we show that the yeast Mediator subunit Gal11p/ Q96RN5 and its activator-targeted KIX domain plays a critical role in fatty acid-dependent transcriptional regulation of fatty acid beta-oxidation and peroxisomal genes by Oaf1p and for the ability of yeast to utilize fatty acids as a sole carbon source . Moreover , structural studies by NMR spectroscopy reveal that the Oaf1p activation domain interacts with the Gal11p/ Q96RN5 KIX domain in a manner similar to the yeast zinc cluster family member and xenobiotic receptor Pdr1p , revealing that the Gal11p/ Q96RN5 KIX domain is a key target of several ligand-dependent transcription factors in yeast . Together with previous work showing that the Caenorhabditis elegans Gal11p/ Q96RN5 homolog MDT-15 plays a critical role in regulation of fatty acid metabolism by the nematode Q07869 -like nuclear receptor NHR-49 , the findings presented here provide evidence for an ancient and essential role of a Mediator co-activator subunit in regulation of fatty acid metabolism by nuclear receptor-like transcription factors in eukaryotes . Discrete roles for peroxisome proliferator-activated receptor gamma and retinoid X receptor in recruiting nuclear receptor coactivators . P37231 ( PPARgamma ) plays a major role in adipogenesis . PPARgamma binds to DNA as a heterodimer with retinoid X receptor ( RXR ) , and PPARgamma-RXR can be activated by ligands specific for either receptor ; the presence of both ligands can result in a cooperative effect on the transactivation of target genes . How these ligands mediate transactivation , however , remains unclear . PPARgamma is known to interact with both the P52701 / Q15788 family of coactivators and the distinct , multisubunit coactivator complex called DRIP . A single DRIP subunit , Q15648 ( Q15648 , PBP ) , binds directly to PPARgamma . Here we report that PPARgamma and RXR selectively interacted with Q15648 and P52701 proteins in a ligand-dependent manner . At physiological concentrations , RXR-specific ligands only induced P52701 binding to RXR , and PPARgamma-specific ligands exclusively recruited Q15648 but not P52701 coactivators to PPARgamma . This selectivity was not observed in interaction assays off DNA , implying that the specificity of coactivator binding in response to ligand is strongly influenced by the allosteric effects of DNA-bound heterodimers . These coactivator-selective effects were also observed in transient-transfection assays in the presence of overexpressed P52701 or DRIP coactivators . The results suggest that the cooperative effects of PPARgamma- and RXR-specific ligands may occur at the level of selective coactivator recruitment . Regulation of angiotensin II receptors beyond the classical pathway . The DB01367 ( renin-angiotensin system ) plays a role not only in the cardiovascular system , including blood pressure regulation , but also in the central nervous system . AngII ( angiotensin II ) binds two major receptors : the AT(1) receptor ( AngII type 1 receptor ) and AT(2) receptor ( AngII type 2 receptor ) . It has been recognized that AT(2) receptor activation not only opposes AT(1) receptor actions , but also has unique effects beyond inhibitory cross-talk with AT(1) receptor signalling . Novel pathways beyond the classical actions of DB01367 , the P12821 ( angiotensin-converting enzyme ) /AngII/AT(1) receptor axis , have been highlighted : the Q9BYF1 /Ang-(1-7) [ angiotensin-(1-7) ] /Mas receptor axis as a new opposing axis against the P12821 /AngII/AT(1) receptor axis , novel AngII-receptor-interacting proteins and various AngII-receptor-activation mechanisms including dimer formation . Q6RW13 ( AT(1)-receptor-associated protein ) and Q9ULD2 ( AT(2)-receptor-interacting protein ) are well-characterized AngII-receptor-associated proteins . These proteins could regulate the functions of AngII receptors and thereby influence various pathophysiological states . Moreover , the possible cross-talk between Q07869 ( peroxisome-proliferator-activated receptor ) -γ and AngII receptor subtypes is an intriguing issue to be addressed in order to understand the roles of DB01367 in the metabolic syndrome , and interestingly some ARBs ( AT(1)-receptor blockers ) have been reported to have an AT(1)-receptor-blocking action with a partial Q07869 -γ agonistic effect . These emerging concepts concerning the regulation of AngII receptors are discussed in the present review . DB01098 , a new P04035 inhibitor , reduces the colonic inflammatory response in dextran sulfate sodium-induced colitis in mice . The aim of the present study was to elucidate the beneficial effects of rosuvastatin , a new P04035 inhibitor , on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium ( DSS ) colitis model . Acute colitis was induced using 8 % DSS in female BALB/c mice . Colonic mucosal inflammation was evaluated clinically , biochemically , and histologically . Mucosal protein contents and mRNA levels of tumor necrosis factor ( P01375 ) -alpha were determined by immunoassay and real time-PCR . The mRNA levels of endothelial nitric oxide synthase ( P29474 ) were determined by real-time PCR . Disease activity scores in DSS-induced colitis model mice , as determined by weight loss , stool consistency , and blood in stool , were significantly lower in the rosuvastatin-treated mice than in control mice . Shortening of the colon was significantly reversed by rosuvastatin . Increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by treatment with rosuvastatin . DB01098 also inhibited increases in intestinal P01375 protein and mRNA expression after DSS administration , respectively . The mucosal mRNA levels of P29474 were decreased after DSS administration , but preserved in mice treated with rosuvastatin . These results suggest that rosuvastatin prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory responses associated with the preservation of P29474 transcription . P37231 regulates P12830 expression and inhibits growth and invasion of prostate cancer . P37231 ( PPARgamma ) might not be permissive to ligand activation in prostate cancer cells . Association of PPARgamma with repressing factors or posttranslational modifications in PPARgamma protein could explain the lack of effect of PPARgamma ligands in a recent randomized clinical trial . Using cells and prostate cancer xenograft mouse models , we demonstrate in this study that a combination treatment using the PPARgamma agonist pioglitazone and the histone deacetylase inhibitor valproic acid is more efficient at inhibiting prostate tumor growth than each individual therapy . We show that the combination treatment impairs the bone-invasive potential of prostate cancer cells in mice . In addition , we demonstrate that expression of P12830 , a protein involved in the control of cell migration and invasion , is highly up-regulated in the presence of valproic acid and pioglitazone . We show that P12830 expression responds only to the combination treatment and not to single PPARgamma agonists , defining a new class of PPARgamma target genes . These results open up new therapeutic perspectives in the treatment of prostate cancer . Disruption of ERalpha signalling pathway by PPARgamma agonists : evidences of PPARgamma-independent events in two hormone-dependent breast cancer cell lines . P37231 ( PPARgamma ) is a nuclear receptor that can be activated by natural ligands such as 15-deoxy-delta(12,14)-prostaglandin J2 ( 15d-PGJ(2) ) as well as synthetic drugs such as thiazolidinediones . The treatment of human breast cancer cell lines with PPARgamma agonists is known to have antiproliferative effects but the role of PPARgamma activation in the process remains unclear . In the present study , we investigated the effects of four PPARgamma agonists , Rosiglitazone ( RGZ ) , DB09201 ( CGZ ) , DB00197 ( Q96PF1 ) and the natural agonist 15d-PGJ(2) , on estrogen receptor alpha ( ERalpha ) signalling pathway in two hormone-dependent breast cancer cell lines , MCF-7 and ZR-75-1 . In both of them , Q96PF1 , CGZ and 15d-PGJ(2) induced an inhibition of ERalpha signalling associated with the proteasomal degradation of ERalpha . ZR-75-1 cells were more sensitive than MCF-7 cells to these compounds . Treatments that induced ERalpha degradation inhibited cell proliferation after 24 h . In contrast , 24 h exposure to RGZ , the most potent activator of PPARgamma disrupted neither ERalpha signalling nor cell proliferation . 9-cis retinoic acid never potentiated the proteasomal degradation of ERalpha . PPARgamma antagonists ( T0070907 , BADGE and GW 9662 ) did not block the proteolysis of ERalpha in MCF-7 and ZR-75-1 cells treated with Q96PF1 . ERalpha proteolysis still occurred in case of PPARgamma silencing as well as in case of treatment with the PPARgamma-inactive compound Delta2- Q96PF1 , demonstrating a PPARgamma-independent mechanism . The use of thiazolidinedione derivatives able to trigger ERalpha degradation by a PPARgamma-independent pathway could be an interesting tool for breast cancer therapy . Ligands of peroxisome proliferator-activated receptor-gamma induce apoptosis in AR42J cells . INTRODUCTION : Peroxisome proliferator-activated receptor-gamma ( P37231 ) is a ligand-activated transcription factor that controls growth , differentiation , and inflammation in different tissues . Roles of P37231 activation in pancreatic acinar cells are poorly characterized . AIMS : To examine the effects of P37231 activation on the induction of apoptosis in rat pancreatic AR42J cells . METHODOLOGY : AR42J cells were treated with ligands of P37231 , and induction of apoptosis was evaluated by cell viability , DNA-fragmentation , and flow cytometry . RESULTS : Treatment of the cells with ligands of P37231 ( 15-deoxy-open triangle12,14-prostaglandin J2 or troglitazone ) induced apoptosis in a dose-dependent manner . DB00197 -induced apoptosis was not blocked by inhibitors of caspases ( acetyl-DEVD-aldehyde and benzoyloxycarbonyl-VAD-fluoromethylketone ) . DB00197 induced the expression of pancreatitis-associated protein-1 and clusterin mRNAs . DB00197 activated c-Jun NH2-terminal kinase/stress-activated protein kinase , but inhibited the activation of extracellular signal-regulated kinases 1/2 . DB00197 did not activate NF-kappaB , suggesting a role of NF-kappaB-independent pathways . In AR42J cells and isolated pancreatic acini , P37231 gene and protein were detected . In addition , troglitazone increased the Q07869 -dependent transcriptional activity , suggesting that P37231 is functional in AR42J cells . CONCLUSION : These results indicate that activation of P37231 induces apoptosis in AR42J cells and imply that P37231 may be a potential therapeutic target of pancreatic inflammation , because of its anti-inflammatory effects in addition to its proapoptotic effects . Cdk5 inhibitor roscovitine alleviates neuropathic pain in the dorsal root ganglia by downregulating N-methyl-D-aspartate receptor subunit 2A . P12004 -dependent kinase 5 ( Cdk5 ) is a member of the small proline-directed serine/threonine kinase family . Cdk5 is not involved in cell cycle regulation , but is implicated in neurodegenerative disorders . However , the role of Cdk5 in neuropathic pain remains unclear . This study aimed to evaluate the possibility that Cdk5 is involved in neuropathic pain in the dorsal root ganglia ( Q86YR7 ) . We injected intrathecally Cdk5 inhibitor roscovitine in rat model of chronic compression of dorsal root ganglion and examined pain behaviors and the expression of N-methyl-d-aspartate receptor subunit 2A ( Q12879 ) but not Q13224 or Q9UHB4 in Q86YR7 . We found that roscovitine alleviated neuropathic pain , causing decline in paw withdrawal mechanical threshold and paw withdrawal thermal latency . Furthermore , roscovitine inhibited Q12879 expression in Q86YR7 . These data suggest that Cdk5- Q12879 pathway regulates neuropathic pain in Q86YR7 , and intrathecal injection of roscovitine could alleviate neuropathic pain . Our findings provide new insight into the analgesic effects of Roscovitine and identify Cdk5- Q12879 pathway as a potential target for effective treatment of neuropathic pain . LG839 : anti-obesity effects and polymorphic gene correlates of reward deficiency syndrome . INTRODUCTION : This study systematically assessed the weight management effects of a novel experimental DNA-customized nutraceutical , LG839 ( LifeGen , Inc. , La Jolla , CA , USA ) . METHODS : A total of 1058 subjects who participated in the overall D.I.E.T. study were genotyped and administered an LG839 variant based on polymorphic outcomes . A subset of 27 self-identified obese subjects of Dutch descent , having the same DNA pattern of four out of the five candidate genes tested ( chi-square analysis ) as the entire data set , was subsequently evaluated . Simple t tests comparing a number of weight management parameters before and after 80 days of treatment with LG839 were performed . RESULTS : Significant results were observed for weight loss , sugar craving reduction , appetite suppression , snack reduction , reduction of late night eating ( all P < 0.01 ) , increased perception of overeating , enhanced quality of sleep , increased happiness ( all P < 0.05 ) , and increased energy ( P < 0.001 ) . Polymorphic correlates were obtained for a number of genes ( P41159 , Q07869 -gamma2 , P42898 , 5- Q13049 , and P14416 genes ) with positive clinical parameters tested in this study . Of all the outcomes and gene polymorphisms , only the P14416 gene polymorphism ( A1 allele ) had a significant Pearson correlation with days on treatment ( r=0.42 , P=0.045 ) . CONCLUSION : If these results are confirmed in additional rigorous , controlled studies , we carefully suggest that DNA-directed targeting of certain regulator genes , along with customized nutraceutical intervention , provides a unique framework and strategic modality to combat obesity . Treatment of cardiovascular dysfunction associated with the metabolic syndrome and type 2 diabetes . Our previous studies have shown vascular dysfunction in small coronary and mesenteric arteries in Zucker obese rats , a model of the metabolic syndrome , and Zucker Diabetic Fatty ( ZDF ) rats , a model of type 2 diabetes . Because of their lipid lowering action and antioxidant activity , we predicted that treatment with DB01098 , an P04035 inhibitor ( statin ) or Enalapril , an angiotensin converting enzyme ( P12821 ) inhibitor would improve vascular dysfunction associated with the metabolic syndrome and type 2 diabetes . METHODS : 20-week-old Zucker obese and 16-week-old ZDF rats were treated with DB01098 ( 25 mg/kg/day ) or Enalapril ( 20 mg/kg/day ) for 12 weeks . We examined metabolic parameters , indices of oxidative stress and vascular dysfunction in ventricular and mesenteric small arteries ( 75-175 microm intraluminal diameter ) from lean , Zucker obese and ZDF rats ( untreated and treated ) . RESULTS : Endothelial dependent responses were attenuated in coronary vessels from Zucker obese and ZDF rats compared to responses from lean rats . Both drugs improved metabolic parameters , oxidative stress , and vascular dysfunction in Zucker obese rats , however , only partial improvement was observed in ZDF rats , suggesting more aggressive treatment is needed when hyperglycemia is involved . CONCLUSION : Vascular dysfunction is improved when Zucker obese and , to a lesser degree , when ZDF rats were treated with DB01098 or Enalapril . P01308 resistance , oxidative stress , and podocyte injury : role of rosuvastatin modulation of filtration barrier injury . BACKGROUND/AIM : There is an emerging relationship between insulin resistance/hyperinsulinemia , oxidative stress , and glomerular injury manifesting as albuminuria . P04035 inhibitors ( statins ) have been shown to reduce oxidative stress in the vasculature as well as albuminuria in animal models and in human studies . The glomerular filtration barrier is emerging as a critical determinant of albumin filtration . We investigated the effects of insulin resistance and rosuvastatin or placebo on the glomerular filtration barrier . METHOD : Young Zucker obese and Zucker lean rats ( 6-7 weeks old ) were treated with the P04035 inhibitor rosuvastatin ( 10 mg/kg/day ) or placebo for 21 days . RESULTS : In the Zucker obese rats , homeostasis model assessment-insulin resistance index , oxidative markers ( NADPH oxidase activity , reactive oxygen species , and urine isoprostane formation ) , podocyte foot process effacement , and albuminuria were increased as compared with Zucker lean controls , independent of increases in systolic blood pressure . Albuminuria correlated with podocyte foot process effacement ( r(2) = 0.61 ) and insulin level ( r(2) = 0.69 ) . DB01098 treatment improved albuminuria , filtration barrier indices , and oxidative stress via copper/zinc superoxide dismutase . CONCLUSIONS : These data indicate that hyperinsulinemia together with insulin resistance is associated with podocyte injury and albuminuria independent of the systolic blood pressure . Further , rosuvastatin modulates filtration barrier injury and albuminuria and improves oxidative stress measures via copper/zinc superoxide dismutase . Therapeutic effects of troglitazone in experimental chronic pancreatitis in mice . Peroxisome proliferator-activated receptor ( Q07869 ) -gamma controls growth , differentiation , and inflammation . P37231 agonists exert anti-inflammatory effects in vitro and inhibit the activation of pancreas stellate cells , implicated in the formation and progression of fibrosis . We determined the influence of troglitazone , a ligand for P37231 , on pancreatic damage and fibrosis in experimental chronic pancreatitis . Mice received six hourly intraperitoneal injections with 50 microg/kg of cerulein or saline , three times a week for 6 weeks . One week after the last injection all mice were sacrificed . Untreated mice were compared with mice treated with troglitazone either during weeks 1 to 6 or weeks 4 to 6 . All mice that received cerulein injections displayed histopathological signs of chronic pancreatitis at week 7 . DB00197 treatment improved all markers for severity of pancreatitis . Moreover , early and postponed troglitazone treatments were equally effective in diminishing intrapancreatic fibrosis as quantified by Sirius red staining , hydroxyproline content , and laminin staining as well as the increased number of pancreatic stellate cells and pancreas levels of transforming growth factor-beta . Thus , troglitazone attenuated pancreatic damage and inflammation in experimental chronic pancreatitis and remained beneficial in a therapeutic setting when given after initial damage had been established . Knockdown of receptor-interacting serine/threonine protein kinase-2 ( O43353 ) affects EMT-associated gene expression in human hepatoma cells . BACKGROUND : Receptor-interacting serine/threonine protein kinase-2 ( O43353 ) has been reported to be an important regulator of tumor proliferation , differentiation and wound repair . We investigated the effects of O43353 knockdown in human hepatoma cells on epithelial-to-mesenchymal transition ( EMT ) -associated gene expression . MATERIALS AND METHODS : HepG2 cells stably expressing O43353 -shRNA ( HepG2-shRIPK2 ) were generated after puromycin selection . Total RNAs from HepG2-shRIPK2 and from HepG2-shcontrol cells were isolated and PCR-based arrays were performed to compare the 84 EMT-associated gene expressions . RESULTS : We observed that knockdown of O43353 down-regulated mRNA expression of jagged 1 ( P78504 ) ; plasminogen activator inhibitor-1 ( P05121 ) ; regulator of G-protein signalling 2 , 24 kDa ( P41220 ) ; P12830 ( CDH1 ) ; fibroblast growth factor binding protein 1 ( Q14512 ) ; snail homolog 2 ( O43623 ) ; protein tyrosine phosphatase type IVA , member 1 ( Q93096 ) ; keratin 19 ( P08727 ) ; vimentin ( P08670 ) ; and survival of motor neuron protein-interacting protein 1 ( SIP1 ) . CONCLUSION : We found that knockdown of O43353 down-regulated nuclear factor kappa B ( NF-κB ) -dependent P05121 and P08670 gene expressions . O43353 might play an important role in hepatic cell migration . These findings could shed new light on carcinogenesis and on liver regeneration . The P28335 receptor agonist lorcaserin reduces nicotine self-administration , discrimination , and reinstatement : relationship to feeding behavior and impulse control . DB04871 ( ( 1R ) -8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine HCl ) is a selective 5-HT(2C) receptor agonist with clinical efficacy in phase-III obesity trials . Based on evidence that this drug class also affects behaviors motivated by drug reinforcement , we compared the effect of lorcaserin on behavior maintained by food and nicotine reinforcement , as well as the stimulant and discriminative stimulus properties of nicotine in the rat . Acutely administered lorcaserin ( 0.3-3 mg/kg , subcutaneous ( SC ) ) dose dependently reduced feeding induced by 22-h food deprivation or palatability . Effects up to 1 mg/kg were consistent with a specific effect on feeding motivation . DB04871 ( 0.6-1 mg/kg , SC ) reduced operant responding for food on progressive and fixed ratio schedules of reinforcement . In this dose range lorcaserin also reversed the motor stimulant effect of nicotine , reduced intravenous self-administration of nicotine , and attenuated the nicotine cue in rats trained to discriminate nicotine from saline . DB04871 also reduced the reinstatement of nicotine-seeking behavior elicited by a compound cue comprising a nicotine prime and conditioned stimulus previously paired with nicotine reinforcement . DB04871 did not reinstate nicotine-seeking behavior or substitute for a nicotine cue . Finally , lorcaserin ( 0.3-1 mg/kg ) reduced nicotine-induced increases in anticipatory responding , a measure of impulsive action , in rats performing the five-choice serial reaction time task . Importantly , these results indicate that lorcaserin , and likely other selective 5-HT(2C) receptor agonists , similarly affect both food- and nicotine-motivated behaviors , and nicotine-induced impulsivity . Collectively , these findings highlight a therapeutic potential for 5-HT(2C) agonists such as lorcaserin beyond obesity into addictive behaviors , such as nicotine dependence . DB00142 reverses Pb2+-induced reductions of DB01221 receptor subunits in vitro . The objective of this study is to determine the effects of Pb2+ on N-methyl-d-aspartate receptor ( NMDAR ) subunits -- Q07869 , Q12879 and Q13224 in primary cultured neuronal cells . We hypothesize that L-glutamic acid ( GA ) reverses Pb2+-induced NMDAR damage . Neuronal cells were isolated from the fetus brain at 18-20th day of gestation of pregnant Sprague Dawley ( SD ) rats . All experiments were included three independent cell preparations ( N=3 ) . The neuronal cells were exposed to Pb2+ ( 10(-10) , 10(-9) , 10(-8) and 10(-7)M ) for 24 h . Neurons were pretreated with NMDAR agonist -- L-glutamic acid ( GA ) ( 200 microM ) and antagonists dizocipine ( MK-801 , 50 nM ) for 1h and then exposed to 10(-7)M of Pb2+ for 24 h . Finally , GA at 2 , 0.2 and 0.02 mM was incubated with neurons prior to Pb2+ exposure . Aliquots of Q9UHB4 , Q12879 and Q13224 proteins from cell homogenate were immunoprecipitated with protein A agarose and detected by Western blotting . The addition of GA unconventionally reversed the reductions of NMDAR by Pb at protein levels , whereas MK-801 exacerbated Pb2+-induced damage . The protection by GA against Pb2+-induced reduction of NMDAR was dose-dependent . These findings suggest that the administration of GA may be a potential approach to intervene the Pb2+-induced NMDAR alterations . Gene expression analysis of troglitazone reveals its impact on multiple pathways in cell culture : a case for in vitro platforms combined with gene expression analysis for early ( idiosyncratic ) toxicity screening . P37231 ( PPARgamma ) agonists of the thiazolidinedione family are used for the treatment of type 2 diabetes mellitus due to their ability to reduce glucose and lipid levels in patients with this disease . Three thiazolidinediones that were approved for treatment are Rezulin ( troglitazone ) , Avandia ( rosiglitazone ) , and Actos ( pioglitazone ) . DB00197 was withdrawn from the market due to idiosyncratic drug toxicity . Rosiglitazone and pioglitazone are still on the market for the treatment of type 2 diabetes . The authors present data from a gene expression screen that compares the impact these three compounds have in rats , in rat hepatocytes , and in the clone 9 rat liver cell line . The authors monitored the changes in expression in multiple genes , including those related to xenobiotic metabolism , proliferation , DNA damage , oxidative stress , apoptosis , and inflammation . Compared to the other two compounds , troglitazone had a significant impact on many of the pathways monitored in vitro although no major perturbation was detected in vivo . The changes detected predict not only general toxicity but potential mechanisms of toxicity . Based on gene expression analysis , the authors propose there is not just one but multiple ways troglitazone could be toxic , depending on a patient 's environment and genetic makeup , including immune response-related toxicity . Sequence variation in P37231 may underlie differential response to troglitazone . Thiazolidinediones ( TZDs ) are peroxisome proliferator-activated receptor-gamma ( P37231 ) agonists used to treat type 2 diabetes . TZDs can also be used to reduce rates of type 2 diabetes in at-risk individuals . However , a large fraction of TZD-treated patients ( 30-40 % ) do not respond to TZD treatment with an improvement in insulin sensitivity ( Si ) . We hypothesized that variation within the gene encoding P37231 may underlie this differential response to TZD therapy . We screened approximately 40 kb of P37231 in 93 nondiabetic Hispanic women ( 63 responders and 30 nonresponders ) with previous gestational diabetes who had participated in the DB00197 In the Prevention Of Diabetes study . TZD nonresponse was defined as the lower tertile in change in Si after 3 months of treatment . Baseline demographic and clinical measures were not different between responders and nonresponders . We identified and genotyped 131 variants including 126 single nucleotide polymorphisms and 5 insertion-deletion polymorphisms . Linkage disequilibrium analysis identified five haplotype blocks . Eight variants were associated with TZD response ( P < 0.05 ) . Three variants were also associated with changes in Si as a continuous variable . Our results suggest that P37231 variation may underlie response to TZD therapy in women at risk for type 2 diabetes . Binding of antipsychotic drugs to human brain receptors focus on newer generation compounds . Using radioligand binding assays and post-mortem normal human brain tissue , we obtained equilibrium dissociation constants ( K(d)s ) for nine new antipsychotic drugs ( iloperidone , melperone , olanzapine , ORG 5222 , quetiapine , risperidone , sertindole , ziprasidone , and zotepine ) , one metabolite of a new drug ( 9-OH-risperidone ) , and three older antipsychotics ( clozapine , haloperidol , and pimozide ) at nine different receptors ( alpha1-adrenergic , alpha2-adrenergic , dopamine D2 , histamine H1 , muscarinic , and serotonin P08908 , P28221 , 5- Q13049 , and P28335 receptors ) . DB04946 was the most potent drug at the two adrenergic receptors . ORG 5222 was the most potent drug at dopamine D2 and 5-HT2c receptors , while ziprasidone was the most potent compound at three serotonergic receptors ( P08908 , P28221 , and 5- Q13049 ) . At the remaining two receptors , olanzapine was the most potent drug at the histamine H1 receptor ( Kd=0.087 nM ) ; clozapine at the muscarinic receptor ( Kd=9 nM ) . Certain therapeutic and adverse effects , as well as certain drug interactions can be predicted from a drug 's potency for blocking a specific receptor . These data can provide guidelines for the clinician in the choice of antipsychotic drug . Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . DB00197 inhibits tumor growth in hepatocellular carcinoma in vitro and in vivo . P37231 ( PPARgamma ) has been implicated in the differentiation and growth inhibition of cancer cells . We examined the effects of PPARgamma activation by troglitazone on hepatocellular carcinoma ( HCC ) cell growth , proliferation , and apoptosis in vitro and in vivo . We also studied relationships between PPARgamma activation and cyclooxygenase-2 ( P35354 ) expression . Human HCC cell lines Huh7 and Hep3B were cultured in the presence or absence of troglitazone . Cell growth was determined via WST-1 assay , proliferation by cell cycle analysis and proliferating cell nuclear antigen ( P12004 ) Western blotting , and apoptosis by flow cytometry and TUNEL . Tumor growth after subcutaneous implantation of Huh7 cells in nude mice was monitored , and the effects of treatment with troglitazone were determined . In resected HCCs , PPARgamma expression was less compared with the histologically normal surrounding liver . In cultures of Hep3B and Huh7 cells , basal expression of PPARgamma was relatively low , but troglitazone caused dose-dependent induction of PPARgamma expression . Cell cycle analysis revealed a decreased proportion of cells in S phase , with arrest at G0/ P55008 . Concomitant downregulation of P12004 and an increase in TUNEL staining , cells were consistent with decreased proliferation and induction of apoptosis by troglitazaone . DB00197 -mediated PPARgamma activation also suppressed P35354 expression and induced p27 in HCC cells . Administration of troglitazone to Huh7 tumor-bearing mice significantly reduced tumor growth and caused tumor regression . In conclusion , collectively , these results indicate that PPARgamma could be a regulator of cell survival and growth in HCC . PPARgamma therefore represents a putative molecular target for chemopreventive therapy or inhibition of liver cancer growth . P37231 ligand prevents the development of chronic pancreatitis through modulating NF-kappaB-dependent proinflammatory cytokine production and pancreatic stellate cell activation . PURPOSE : Thiazolidinedione derivatives ( TZDs ) are known to be ligands of peroxisome proliferator-activated receptor gamma ( PPARgamma ) . In this study , we investigated the effect of a TZD , troglitazone , on inflammation and fibrogenesis in the pancreas of an experimental model of chronic pancreatitis . MATERIAL AND METHODS : Male WBN/Kob rats with spontaneous chronic pancreatitis were fed rat chow containing 0.2 % troglitazone from 1 to 4 months of age . Immunohistochemical studies of rat pancreas were carried out with monoclonal mouse antibody against human alpha-smooth muscle actin ( alpha-SMA ) or rabbit polyclonal antibody against collagen type I , collagen type III , or fibronectin . Cytokine production was measured by enzyme-linked immunosorbent assay . The inhibitory action of troglitazone on nuclear factor-kappaB ( NF-kappaB ) binding activity in activated macrophages was also investigated . RESULTS : Long-term administration of troglitazone reduced inflammatory cell infiltration and fibrosis in the pancreas of WBN/Kob rats , and expression of alpha-SMA , procollagen I , III , and fibronectin was significantly reduced by troglitazone . The increase in P01375 production by activated macrophages was significantly decreased by troglitazone . Peritoneal macrophages isolated from WBN/Kob rats produced a large amount of P01375 , whereas those from troglitazone-treated WBN/Kob rats produced only a marginal amount of P01375 . Lipopolysaccharide-induced NF-kappaB binding activity in peritoneal macrophages was also significantly reduced by troglitazone . CONCLUSIONS : DB00197 prevented the progression of chronic pancreatitis via inhibition of Q13201 synthesis and proinflammatory cytokine production mediated by the inhibition of NF-kappaB activity . Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 -R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 however exhibited an altered binding activity in cancer specimens . DB04839 did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event . DB00197 , a P37231 activator prevents endothelial cell adhesion molecule expression and lymphocyte adhesion mediated by P01375 . BACKGROUND : Cytokine mediated induction of the mucosal addressin cell adhesion molecule-1( Q13477 ) expression is associated with the onset and progression of inflammatory bowel disease ( Q9UKU7 ) . RESULTS : Using western blotting and cell-based ELISA , we show in this study that troglitazone , an activator of the peroxisome proliferator-activated receptor-gamma ( P37231 ) , widely used in the treatment of diabetes , has as well recently been highlighted as protective in models of inflammation and cancer . We found that troglitazone ( 10-40 microM ) , significantly reduced the P01375 ( 1 ng/ml ) mediated induction of endothelial Q13477 in a dose-dependent manner , achieving a 34.7 % to 98.4 % reduction in induced Q13477 . Trogliazone ( 20 microM ) reduced P01375 induced P19320 , P05362 and P16581 expression . Moreover , troglitazone significantly reduced alpha4beta7-integrin dependent lymphocyte adhesion to P01375 cultured endothelial cells . CONCLUSIONS : These results suggest that P37231 agonists like troglitazone may be useful in the clinical treatment of Q9UKU7 . Novel function of androgen receptor-associated protein 55/ O43294 as a negative regulator of P84022 signaling . P10275 -associated protein 55 ( O43294 / O43294 ) belongs to the LIM protein superfamily and is featured by three or four N-terminal LD motifs and four C-terminal zinc finger-like LIM domains . Both LD motifs and LIM domains can serve as protein-protein interaction interfaces . Recently , we found that enforced expression of O43294 inhibits transforming growth factor-beta-mediated up-regulation of Smad binding element-luciferase reporter activity in NRP-154 and NRP-152 rat prostate and LNCaP human prostate cell lines . Moreover , O43294 also inhibits the induction of Smad-binding element 4-luciferase and 3TP-luciferase ( a plasminogen activator inhibitor-1 ( P05121 ) promoter construct ) reporters by constitutively active ( CA ) - P84022 in these cell lines . Co-immunoprecipitation studies suggest an interaction between O43294 and either CA- P84022 or wild-type P84022 in HEK293 cells that occurs through the MH2 domain of P84022 and the C terminus of O43294 with wild-type P84022 having stronger affinity than CA- P84022 to O43294 . O60760 pull-down assays demonstrate that this interaction can occur in a cell-free system . These results are consistent with the luciferase data showing that the C terminus of O43294 is critical for suppression of P84022 activity . Furthermore , using a mammalian two-hybrid system , we confirmed that O43294 interacts with the MH2 domain of P84022 and suppresses CA- P84022 -induced transcriptional responses . In conclusion , these results support that O43294 selectively intercepts transforming growth factor-beta signaling through an interaction of the LIM domain of O43294 with the MH2 domain of P84022 . The role of tumor suppressor dysregulation in prostate cancer progression . P10275 activity is essential for prostate cancer development and progression . While there are classically defined roles for the retinoblastoma ( P06400 ) and p53 tumor suppressor pathways in maintenance of cell cycle control and the DNA damage response , recent studies have demonstrated a direct role of these two pathways in regulating AR expression and function . While the role of Pten deregulation in prostate cancer has provided much insight in to the mechanisms underlying prostate cancer initiation and progression , emerging roles for P06400 and p53 are likely to further expand upon our understanding of tumor suppressor/nuclear receptor interaction . As disconnecting mitogenic signaling from AR-mediated gene transcription underlies the progression to castrate resistant prostate cancer ( CRPC ) , functional inactivation of these two tumor suppressor pathways represents one mechanism through which AR protein levels can be upregulated and AR-mediated gene transcription can become aberrant . Importantly , recent advances in small molecule inhibitor design and discovery have led to the identification of agents capable of targeting these two prominent pathways and restoring the function of deregulated wild-type P06400 and p53 protein . While such agents have undergone extensive study in many solid tumor types , the additional importance of P06400 and p53 in restraining transcription of the AR gene within the prostate provides impetus for examining how loss of these two tumor suppressor proteins can facilitate transition of prostate cancers to CRPC . As will be reviewed in this article , restoration of P06400 and p53 functions are not only important in regard to shortterm cell cycle regulation and response to genomic stresses , but likely have direct implications for deregulation of the AR locus . Coadministration of acetaminophen and troglitazone : pharmacokinetics and safety . DB00197 , a P37231 agonist , enhances the actions of insulin on muscle and liver . It is metabolized predominantly in the liver to a sulfate conjugate and a quinone metabolite . Acetaminophen also undergoes metabolism by conjugation . This three-way crossover study in 12 healthy male volunteers was conducted to investigate the effects of acetaminophen on the metabolism of troglitazone and vice versa . No statistically or clinically relevant differences in area under the concentration-time curve extrapolated to infinity ( AUCinfinity ) were observed for troglitazone , its quinone metabolite , or acetaminophen . A statistically significant decrease in troglitazone sulfate conjugate during administration with acetaminophen was not clinically relevant . No statistically or clinically relevant differences were observed in maximum concentration ( Cmax ) , time to Cmax ( tmax ) , or elimination half-life of troglitazone , its two main metabolites , and acetaminophen or in acetaminophen urinary sulfate excretion , although there was a slight decrease in acetaminophen glucuronide excretion during administration with troglitazone . Adverse events were minor and similar between treatments . These findings suggest that troglitazone and acetaminophen can be coadministered without adverse clinical consequences . P06401 level as a predictor of response to megestrol acetate in advanced breast cancer : a retrospective study . DB00351 ( 160 mg/day ) produced a response rate of 44 % in a retrospective series of 39 evaluable patients with advanced breast cancer . The estrogen-receptor ( ER ) level was greater than 10 fmols/mg of protein in 28 patients , and the progesterone-receptor ( PR ) level was greater than 10 fmols/mg of protein in 26 patients . ER and PR levels , age , and disease-free interval were analyzed for their relationship to response . The PR was the single best predictor of response to megestrol acetate ; the addition of ER added 2 % to the predictive accuracy rate of PR alone . Influence of labor on neonatal neutrophil apoptosis , and inflammatory activity . Neutrophil apoptosis is impaired in neonates , and this contributes to prolonged inflammation and tissue injury in infants after infection or trauma . In the present studies , we investigated whether labor generates mediators that further suppress apoptosis . We found that neutrophil apoptosis was reduced in neonates exposed to labor , when compared with infants delivered by cesarean section before labor . This was not due to alterations in caspase-3 or inhibitor of apoptosis protein-2 ( IAP-2 ) . In contrast , labor primed neutrophils to express tumor necrosis factor alpha ( P01375 ) , suggesting that proinflammatory mediators contribute to reduced apoptosis after labor . Eicosanoids generated via cyclooxygenase-2 ( Cox-2 ) and lipoxygenase ( Lox ) also regulate neutrophil apoptosis . 15-Lox , which generates proapoptotic lipoxins , but not Cox-2 , was greater in neutrophils before labor , relative to cells exposed to labor . Anti-inflammatory eicosanoids exert their effects in part via peroxisome proliferator-activated receptor gamma ( P37231 ) . Expression of gelatinase-associated lipocalin and catalase , two markers of P37231 activity , were increased in neonatal neutrophils before labor , relative to cells exposed to labor . These findings suggest that the anti-inflammatory environment is maintained before labor , in part , by eicosanoids . Although increased neutrophil longevity after labor is important for host defense in the immediate newborn period , it may contribute to inflammatory or oxidative injury in susceptible infants . Effects of the PPARgamma agonist troglitazone on endothelial cells in vivo and in vitro : differences between human and mouse . P37231 ( PPARgamma ) agonists and PPARgamma/alpha dual agonists have been or are being developed for clinical use in the treatment of type 2 diabetes mellitus and hyperlipidemias . A common tumor finding in rodent carcinogenicity studies for these agonists is hemangioma/hemangiosarcoma in mice but not in rats . We hypothesized that increased endothelial cell proliferation may be involved in the mechanism of Q07869 agonist-induced vascular tumors in mice , and we investigated the effects on endothelial cells utilizing troglitazone , the first clinically used PPARgamma agonist , in vivo and in vitro . DB00197 ( 400 and 800 mg/kg/day ) induced hemangiosarcomas in mice in a 2-year bioassay . We showed that troglitazone increased endothelial cell proliferation in brown and white adipose tissue and liver in mice at sarcomagenic doses after 4 weeks of treatment . DB00197 was cytotoxic both to human dermal microvascular endothelial cells ( HMEC1 ) and mouse mammary fat pad microvascular endothelial cells ( MFP MVEC ) at high concentrations . However , MFP MVEC were more resistant to the cytotoxic effects of troglitazone based on the much lower LC(50) in HMEC1 ( 17.4 muM ) compared to MFP MVEC ( 92.2 muM ) . DB00197 increased the proliferation and survival of MFP MVEC but not HMEC1 in growth factor reduced conditions . Our data demonstrate that troglitazone may induce hemangiosarcomas in mice , at least in part , through enhancement of survival and proliferation of microvascular endothelial cells . Such an effect does not occur with human cells , suggesting that human may react differently to exposure to Q07869 agonists compared with mice . Cardioprotective mechanism of telmisartan via P37231 - P29474 pathway in dahl salt-sensitive hypertensive rats . BACKGROUND : Recently , some investigators have shown that telmisartan , an angiotensin II ( Ang II ) -receptor blocker ( ARB ) , is a partial agonist of the peroxisome proliferator-activated receptor-gamma ( P37231 ) . We investigate whether telmisartan improves cardiovascular remodeling associated with the production of endothelial nitric oxide synthase ( P29474 ) through P37231 , inhibits the Rho-kinase pathway , and suppresses oxidative stress in Dahl salt-sensitive ( DS ) hypertensive rats . METHODS : Telmisartan ( 1 mg/kg per day ) or telmisartan plus P37231 inhibitor , GW9662 ( 1 mg/kg per day ) was administered from the age of 6-11 weeks . Age-matched male Dahl salt-resistant ( DR ) rats served as a control group . RESULTS : The levels of P29474 and P37231 expression , and P29474 phosphorylation were significantly lower in DS rats than in DR rats . Chronic telmisartan treatment in DS rats significantly increased these parameters , but not telmisartan plus GW9662 . Telmisartan effectively inhibited the vascular lesion formation such as medial thickness and perivascular fibrosis , but not telmisartan plus GW9662 . Moreover , upregulated RhoA protein , Rho-kinase mRNA , and myosin light-chain phosphorylation in DS rats was decreased by telmisartan to a similar degree as observed after treatment with Y-27632 , a selective Rho-kinase inhibitor . In addition , NAD(P)H oxidase P13498 , p47phox , gp91phox expression , and mitogen-activated protein kinase and its downstream effector P08133 S6 kinase phosphorylation in DS rats was also inhibited by telmisartan . CONCLUSIONS : These results suggest that the cardioprotective mechanism of telmisartan may be partly due to improvement of endothelial function associated with P37231 - P29474 , oxidative stress , and Rho-kinase pathway . Concomitant loss of p120-catenin and β-catenin membrane expression and oral carcinoma progression with P12830 reduction . The binding of p120-catenin and β-catenin to the cytoplasmic domain of P12830 establishes epithelial cell-cell adhesion . Reduction and loss of catenin expression degrades P12830 -mediated carcinoma cell-cell adhesion and causes carcinomas to progress into aggressive states . Since both catenins are differentially regulated and play distinct roles when they dissociate from P12830 , evaluation of their expression , subcellular localization and the correlation with P12830 expression are important subjects . However , the same analyses are not readily performed on squamous cell carcinomas in which P12830 expression determines the disease progression . In the present study , we examined expression and subcellular localization of p120-catenin and β-catenin in oral carcinomas ( n = 67 ) and its implications in the carcinoma progression and P12830 expression using immunohitochemistry . At the invasive front , catenin-membrane-positive carcinoma cells were decreased in the dedifferentiated ( p120-catenin , P < 0.05 ; β-catenin , P < 0.05 ) and invasive carcinomas ( p120-catenin , P < 0.01 ; β-catenin , P < 0.05 ) and with the P12830 staining ( p120-catenin , P < 0.01 ; β-catenin , P < 0.01 ) . Carcinoma cells with β-catenin cytoplasmic and/or nuclear staining were increased at the invasive front compared to the center of tumors ( P < 0.01 ) . Although the p120-catenin isoform shift from three to one associates with carcinoma progression , it was not observed after TGF-β , P01133 or P01375 -α treatments . The total amount of p120-catenin expression was decreased upon co-treatment of TGF-β with P01133 or P01375 -α . The above data indicate that catenin membrane staining is a primary determinant for P12830 -mediated cell-cell adhesion and progression of oral carcinomas . Furthermore , it suggests that loss of p120-catenin expression and cytoplasmic localization of β-catenin fine-tune the carcinoma progression .

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The effect of 1,25-dihydroxyvitamin D3 on lymphoma cell lines and expression of vitamin D receptor in lymphoma . 1,25(OH)2D3 promotes differentiation and has an antiproliferative effect in a variety of cell lines derived from the immunohaematopoetic system . alpha-Calcidol which is metabolised to 1,25(OH)2D3 has been shown to produce tumour regression in follicular low grade non-Hodgkin 's lymphoma ( Q9NZ71 ) and the dose limiting toxicity is hypercalcaemia . The cellular action of 1,25(OH)2D3 is mediated by binding to an intracellular protein , the vitamin D receptor ( P11473 ) . We have evaluated the activity of 1,25(OH)2D3 and its non-calcaemogenic analogue MC903 in the SU-DHL4 and SU-DUL5 B cell lines which carry the 14;18 translocation characteristic of follicular Q9NZ71 , and also the expression of the P11473 in a range of B cell NHLs . Both agents induced differentiation and had an antiproliferative effect on the SU-DHL4 and SU-DUL5 cell lines . However this occurred at a relatively high concentration ( 10(-7) M ) which exceeds the physiological concentration of 1,25(OH)2D3 by approximately 10(3)-10(4)-fold . Expression of the P11473 was low in each cell line and in the low grade lymphoma tumour samples . To account for the observed clinical response to 1 alpha OHD3 ( alpha-calcidol ) in follicular Q9NZ71 a network is suggested whereby 1,25(OH)2D3 modulates the activity of P01730 +T cells which have previously been shown to promote follicle centre cell proliferation . DB00169 analogues may enable serum levels to be achieved which produce a direct action on follicular lymphoma cells without disturbing calcium metabolism . HIV entry inhibitors : mechanisms of action and resistance pathways . Entry inhibitors represent a new generation of antivirals for the treatment of HIV infection . Several compounds which block the attachment of HIV gp120 to either the P01730 T cell receptor or the P51681 / P61073 co-receptors are currently in clinical development . Most of these compounds have different molecular structures and specific mechanisms of action . These agents are eagerly awaited by a growing number of patients carrying viruses resistant viruses to many of the current available reverse transcriptase and protease inhibitors . For enfuvirtide , the first and , so far , only entry inhibitor approved for clinical use , the main mechanism of resistance is the selection of changes within a 10 amino acid segment encompassing residues 36-45 within the Q96GN5 region of gp41 . For other entry inhibitors , multiple changes in different gp120 domains ( V1 , V2 , V3 , P06681 and C4 ) have been associated with loss of susceptibility to these agents , although in most cases with limited cross-resistance . Ca2+/calmodulin-dependent kinase II signalling cascade mediates Q99572 receptor-dependent inhibition of neuritogenesis in neuroblastoma cells . DB00171 , via purinergic P2X receptors , acts as a neurotransmitter and modulator in both the central and peripheral nervous systems , and is also involved in many biological processes , including cell proliferation , differentiation and apoptosis . Previously , we have reported that Q99572 receptor inhibition promotes axonal growth and branching in cultured hippocampal neurons . In this article , we demonstrate that the Q99572 receptor negatively regulates neurite formation in mouse Neuro-2a neuroblastoma cells through a Ca2+/calmodulin-dependent kinase II-related mechanism . Using both molecular and immunocytochemical techniques , we characterized the presence of endogenous P51575 , P56373 , Q99571 and Q99572 subunits in these cells . Of these , the Q99572 receptor was the only functional receptor , as its activation induced intracellular calcium increments similar to those observed in primary neuronal cultures , exhibiting pharmacological properties characteristic of homomeric Q99572 receptors . Patch-clamp experiments were also conducted to fully demonstrate that ionotropic Q99572 receptors mediate nonselective cation currents in this cell line . Pharmacological inhibition of the Q99572 receptor and its knockdown by small hairpin RNA interference resulted in increased neuritogenesis in cells cultured in low serum-containing medium , whereas Q99572 overexpression significantly reduced the formation of neurites . Interestingly , Q99572 receptor inhibition also modified the phosphorylation state of focal adhesion kinase , Akt and glycogen synthase kinase 3 , protein kinases that participate in the Ca2+/calmodulin-dependent kinase II signalling cascade and that have been related to neuronal differentiation and axonal growth . Taken together , our results provide the first mechanistic insight into Q99572 receptor-triggered signalling pathways that regulate neurite formation in neuroblastoma cells . Neutrophil apoptosis : selective regulation by different ligands of integrin alphaMbeta2 . Neutrophils undergo spontaneous apoptosis , but their survival can be extended during inflammatory responses . alpha(M)beta(2) is reported either to delay or accelerate neutrophil apoptosis , but the mechanisms by which this integrin can support such diametrically opposed responses are poorly understood . The abilities of closely related alpha(M)beta(2) ligands , plasminogen and angiostatin , derived from plasminogen , as well as fibrinogen and its two derivative alpha(M)beta(2) recognition peptides , P1 and P2-C , differed markedly in their effects on neutrophil apoptosis . P00747 , fibrinogen , and P2-C suppressed apoptosis via activation of Akt and P27361 /2 kinases , while angiostatin and P1 failed to activate these prosurvival pathways and did not prevent neutrophil apoptosis . Using cells transfected with alpha(M)beta(2) or its individual alpha(M) or beta(2) subunits , and purified receptors and its constituent chains , we show that engagement of both subunits with prosurvival ligands is essential for induction of the prosurvival response . Hence , engagement of a single integrin by closely related ligands can induce distinct signaling pathways , which can elicit distinct cellular responses . Vitamin D and phosphate regulate fibroblast growth factor-23 in K-562 cells . Fibroblast growth factor-23 ( Q9GZV9 ) has been recently identified as playing an important pathophysiological role in phosphate homeostasis and vitamin D metabolism . To elucidate the precise physiological regulation of Q9GZV9 , we characterized the mouse Q9GZV9 5'-flanking region and analyzed its promoter activity . The 5'-flanking region of the mouse Q9GZV9 gene contained a P20226 site ( TATA box ) and several putative transcription factor binding sites , including P28698 , P15976 and c-Ets-1 motifs , but it did not contain the typical sequences of the vitamin D response element . Plasmids encoding 554-bp ( pGL/-0.6 ) , 364-bp ( pGL/-0.4 ) and 200-bp ( pGL/-0.13 ) promoter regions containing the P20226 element and +1-bp fragments drove the downstream expression of a luciferase reporter gene in transfection assays . We also found that Q9GZV9 mRNA was expressed in K-562 erythroleukemia cell lines but not in MC3T3-E1 , Raji , or Hep G2 human carcinoma cells . Treatment with 1,25-dihydroxyvitamin D3 in the presence of high phosphate markedly stimulated pGL/-0.6 activity , but calcium had no effect . In addition , the plasma Q9GZV9 levels were affected by the dietary and plasma inorganic phosphate concentrations . Finally , the levels of plasma Q9GZV9 in vitamin D receptor-null mice were significantly lower than in wild-type mice . The presents study demonstrated that vitamin D and the plasma phosphate level are important regulators of the transcription of the mouse Q9GZV9 gene . MAPK inhibition by 1alpha,25(OH)2- DB00169 in breast cancer cells . Evidence on the participation of the P11473 and Src . 1alpha,25-Dihydroxyvitamin D(3) [ 1alpha,25(OH)(2)D(3) ] , the hormonally active form of Vitamin D(3) , has been shown to be a potent negative growth regulator of breast cancer cells both in vitro and in vivo . 1alpha,25(OH)(2)D(3) acts through two different mechanisms . In addition to regulating gene transcription via its specific intracellular receptor ( Vitamin D receptor , P11473 ) , 1alpha,25(OH)(2)D(3) induces , rapid , non-transcriptional responses involving activation of transmembrane signal transduction pathways . The mechanisms that mediate the antiproliferative effects of 1alpha,25(OH)(2)D(3) in breast cancer cells are not fully understood . Particularly , there is no information about the early non-genomic signal transduction effectors modulated by the hormone . The present study shows that 1alpha,25(OH)(2)D(3) rapidly inhibits serum induced activation of P27361 and P28482 Q96HU1 kinases . The non-receptor tyrosine kinase Src is involved in the pathway leading to activation of P29323 1/2 by serum . Furthermore , 1alpha,25(OH)(2)D(3) increases the tyrosine-phosphorylated state of Src as well as it inhibits its kinase activity and induces the association of the P11473 with Src . These data suggest that 1alpha,25(OH)(2)D(3) inhibits MAPK by inactivating Src tyrosine kinase through a so far unknown mechanism that seems to be mediated by the P11473 . Differential ligand-dependent interactions between the AF-2 activating domain of nuclear receptors and the putative transcriptional intermediary factors mSUG1 and O15164 . Using a yeast two-hybrid system we report the isolation of a novel mouse protein , mSUG1 , that interacts with retinoic acid receptor alpha ( RAR alpha ) both in yeast cells and in vitro in a ligand- and AF-2 activating domain ( AF-2 AD ) -dependent manner and show that it is a structural and functional homologue of the essential yeast protein P62195 . mSUG1 also efficiently interacts with other nuclear receptors , including oestrogen ( ER ) , thyroid hormone ( TR ) , DB00169 ( P11473 ) and retinoid X ( RXR ) receptors . By comparing the interaction properties of these receptors with mSUG1 and O15164 , we demonstrate that : ( i ) RXR alpha efficiently interacts with O15164 , but not with mSUG1 , whereas TR alpha interacts much more efficiently with mSUG1 than with O15164 , and RAR alpha , P11473 and ER efficiently interact with mSUG1 and O15164 ; ( ii ) the amphipathic alpha-helix core of the AF-2 AD is differentially involved in interactions of RAR alpha with mSUG1 and O15164 ; ( iii ) the AF-2 AD cores of RAR alpha and ER are similarly involved in their interaction with O15164 , but not with mSUG1 . Thus , the interaction interfaces between the different receptors and either mSUG1 or O15164 may vary depending on the nature of the receptor and the putative mediator of its AF-2 function . We discuss the possibility that mSUG1 and O15164 may mediate the transcriptional activity of the AF-2 of nuclear receptors through different mechanisms . Phosphorylation at serine 208 of the 1alpha,25-dihydroxy P11473 modulates the interaction with transcriptional coactivators . Upon ligand binding the 1alpha,25-dihydroxy P11473 ( P11473 ) undergoes a conformational change that allows interaction with coactivator proteins including P52701 / P12931 family members and the multimeric DRIP complex through the Q15648 subunit . Casein kinase II ( CKII ) phosphorylates P11473 both in vitro and in vivo at serine 208 within the hinge domain . This phosphorylation does not affect the ability of P11473 to bind DNA , but increases its ability to transactivate target promoters . Here , we have analyzed whether phosphorylation of P11473 by CKII modulates the ability of P11473 to interact with coactivators in vitro . We find that both mutation of serine 208 to aspartic acid ( VDRS208D ) or phosphorylation of P11473 by CKII enhance the interaction of P11473 with Q15648 in the presence of 1alpha,25-dihydroxy DB00169 . We also find that the mutation VDRS208D neither affects the ability of this protein to bind DNA nor to interact with Q15788 and RXRalpha . Together , our results indicate that phosphorylation of P11473 at serine 208 contributes to modulate the affinity of P11473 for the DRIP complex and therefore may have a role in vivo regulating P11473 -mediated transcriptional enhancement . P22681 mutation-related patterns of phosphorylation and sensitivity to tyrosine kinase inhibitors . Recurrent homozygous P22681 -inactivating mutations in myeloid malignancies decrease ubiquitin ligase activity that inactivates P12931 family kinases ( SFK ) and receptor tyrosine kinases ( RTK ) . However , the most important SFK and RTK affected by these mutations , and hence , the most important therapeutic targets , have not been clearly characterized . We compared SFK and RTK pathway activity and inhibitors in acute myeloid leukemia cell lines containing homozygous R420Q mutation ( GDM-1 ) , heterozygous deletion ( MOLM13 ) and wild-type ( WT ) P22681 ( THP1 , U937 ) . As expected with P22681 loss , GDM-1 displayed high P10721 expression and granulocyte-macrophage colony-stimulating factor ( GM- P04141 ) hypersensitivity . Ectopic expression of WT P22681 decreased GDM-1 proliferation but not cell lines with WT P22681 . GDM-1 , but not the other cell lines , was highly sensitive to growth inhibition by dasatinib ( dual SFK and RTK inhibitor , LD50 50 nM ) ; there was less or no selective inhibition of GDM-1 growth by sunitinib ( RTK inhibitor ) , imatinib ( P00519 , P10721 inhibitor ) , or Q99463 ( SFK inhibitor ) . Phosphoprotein analysis identified phosphorylation targets uniquely inhibited by dasatinib treatment of GDM-1 , including a number of proteins in the P10721 and GM- P04141 receptor pathways ( for example , P10721 Tyr721 , P40763 Tyr705 ) . In conclusion , the promiscuous effects of P22681 loss on SFK and RTK signaling appear to be best targeted by dual SFK and RTK inhibition . Q99572 receptor-dependent intestinal afferent hypersensitivity in a mouse model of postinfectious irritable bowel syndrome . The DB00171 -gated P2X(7) receptor ( P2X(7)R ) was shown to be an important mediator of inflammation and inflammatory pain through its regulation of IL-1β processing and release . Trichinella spiralis-infected mice develop a postinflammatory visceral hypersensitivity that is reminiscent of the clinical features associated with postinfectious irritable bowel syndrome . In this study , we used P2X(7)R knockout mice ( P2X(7)R(-/-) ) to investigate the role of P2X(7)R activation in the in vivo production of IL-1β and the development of postinflammatory visceral hypersensitivity in the T. spiralis-infected mouse . During acute nematode infection , IL-1β-containing cells and P2X(7)R expression were increased in the jejunum of wild-type ( WT ) mice . Peritoneal and serum IL-1β levels were also increased , which was indicative of elevated IL-1β release . However , in the P2X(7)R(-/-) animals , we found that infection had no effect upon intracellular , plasma , or peritoneal IL-1β levels . Conversely , infection augmented peritoneal P01375 -α levels in both WT and P2X(7)R(-/-) animals . Infection was also associated with a P2X(7)R-dependent increase in extracellular peritoneal lactate dehydrogenase , and it triggered immunological changes in both strains . Jejunal afferent fiber mechanosensitivity was assessed in uninfected and postinfected WT and P2X(7)R(-/-) animals . Postinfected WT animals developed an augmented afferent fiber response to mechanical stimuli ; however , this did not develop in postinfected P2X(7)R(-/-) animals . Therefore , our results demonstrated that P2X(7)Rs play a pivotal role in intestinal inflammation and are a trigger for the development of visceral hypersensitivity . DNA methylation-related vitamin D receptor insensitivity in breast cancer . Calcitriol ( 1α , 25(OH)(2)- DB00169 ) binds to the vitamin D receptor ( P11473 ) and regulates differentiation of the normal mammary gland , and may therefore be useful in breast cancer treatment or prevention . Many breast cancer cells are , however , resistant to Calcitriol . In this study , we investigated the resistance mechanism and the role of epigenetic silencing of P11473 by promoter hypermethylation . Bisulfite sequencing of the P11473 promoter region revealed methylated CpG islands at -700 base pairs ( bp ) upstream and near the transcription start site . P11473 CpG islands were demethylated by 5'deoxy-azacytidine treatment , and this was accompanied by a parallel increase in P11473 mRNA levels in breast cancer cell lines . Quantitative methylation-specific PCR analyses confirmed hypermethylation of these CpG islands in primary tumors , and its absence in normal breast tissue . P11473 transcripts detected in breast cancers were predominantly 5'-truncated , while normal breast tissue expressed full-length transcripts . Consistent with this observation , genes containing the P11473 -responsive element ( VDRE ) , such as cytochrome p450 hydroxylases , P38936 or C/EBP were underexpressed in breast cancers compared to normal breast samples . Expression of the active longer transcripts of P11473 was restored with 5'deoxy- DB00928 ( AZA ) treatment , with a concurrent increase in expression of VDRE-containing genes . Thus , promoter methylation-mediated silencing of expression of the functional variants of P11473 may contribute to reduced expression of downstream effectors of the P11473 pathway and subsequent Calcitriol insensitivity in breast cancer . These data suggest that pharmacological reversal of P11473 methylation may re-establish breast cancer cell susceptibility to differentiation therapy using Calcitriol . DB00169 and its nuclear receptor increase the expression and activity of the human proton-coupled folate transporter . Folates are essential for nucleic acid synthesis and are particularly required in rapidly proliferating tissues , such as intestinal epithelium and hemopoietic cells . Availability of dietary folates is determined by their absorption across the intestinal epithelium , mediated by the proton-coupled folate transporter ( Q96NT5 ) at the apical enterocyte membranes . Whereas transport properties of Q96NT5 are well characterized , regulation of Q96NT5 gene expression remains less elucidated . We have studied the mechanisms that regulate Q96NT5 promoter activity and expression in intestine-derived cells . Q96NT5 mRNA levels are increased in Caco-2 cells treated with 1,25-dihydroxyvitamin D(3) ( vitamin D(3) ) in a dose-dependent fashion , and the duodenal rat Pcft mRNA expression is induced by vitamin D(3) ex vivo . The Q96NT5 promoter region is transactivated by the vitamin D receptor ( P11473 ) and its heterodimeric partner retinoid X receptor-alpha ( RXRalpha ) in the presence of vitamin D(3) . In silico analyses predicted a P11473 response element ( VDRE ) in the Q96NT5 promoter region -1694/-1680 . DNA binding assays showed direct and specific binding of the P11473 :RXRalpha heterodimer to the Q96NT5 (-1694/-1680) , and chromatin immunoprecipitations verified that this interaction occurs within living cells . Mutational promoter analyses confirmed that the Q96NT5 (-1694/-1680) motif mediates a transcriptional response to vitamin D(3) . In functional support of this regulatory mechanism , treatment with vitamin D(3) significantly increased the uptake of [(3)H]folic acid into Caco-2 cells at pH 5.5 . In conclusion , vitamin D(3) and P11473 increase intestinal Q96NT5 expression , resulting in enhanced cellular folate uptake . Pharmacological treatment of patients with vitamin D(3) may have the added therapeutic benefit of enhancing the intestinal absorption of folates . Dysregulation of vitamin D3 synthesis leads to enhanced cholangiocarcinoma growth . BACKGROUND : Cholangiocarcinoma is a deadly biliary tumour with limited treatment strategies . Vitamin ( 1,25(OH)2D ) has anti-proliferative effects on several cancers . DB00169 is synthesized by the enzyme , O15528 , and signals via the nuclear vitamin D3 receptor . The enzyme , Q07973 , degrades vitamin D3 . AIMS : ( i ) Measure the expression of O15528 , Q07973 , and vitamin D3 receptor in human nonmalignant and cholangiocarcinoma lines and biopsy control or tumour samples ; and ( ii ) evaluate the effects of vitamin D3 on vitamin D3 synthesis and cholangiocarcinoma growth . METHODS : In vitro studies were performed in malignant and nonmalignant cholangiocytes . P11473 , Q07973 and Q02318 expression was measured in cell lines and biopsy samples . Cell lines were stimulated with vehicle or vitamin D3 from 30min to 48h . Cell viability was assessed by MTS assays and BrdU incorporation . P11473 , Q07973 and O15528 expression was measured in cholangiocarcinoma cells stimulated with vehicle or vitamin D3 . RESULTS : In cholangiocarcinoma lines and biopsy samples , vitamin D3 receptor and Q07973 expression increased compared to controls , whereas O15528 expression was decreased or unchanged . DB00169 induced nuclear translocation of vitamin D3 receptor in cholangiocarcinoma and decreased cholangiocarcinoma growth . CONCLUSION : Treatment with vitamin D3 decreased Q07973 , whereas O15528 expression increased . Modulation of vitamin D3 synthesis may be important in the management of cholangiocarcinoma . Changes in circulating biomarkers during a single hemodialysis session . The hemodialysis ( HD ) procedure induces an inflammatory response potentially contributing to cardiovascular disease . Here we investigated the acute impact of HD on circulating biomarkers . Circulating biomarkers ( small solutes , middle molecular-sized peptides , and proteins ) related to inflammation , oxidative stress , and vascular calcification ( VC ) were measured before and after a single session of HD in 45 clinically stable patients . Concentrations were corrected for ultrafiltration-induced hemoconcentration . Among vascular calcification-related biomarkers , osteoprotegerin and fetuin-A remained unchanged while fibroblast growth factor-23 ( Q9GZV9 ) decreased by -19 % . Changes of Q9GZV9 and changes of phosphate correlated ( ρ = 0.61 , P < 0.001 ) . While P02741 did not change , interleukin-6 ( P05231 ) increased by 14 % and pentraxin 3 ( PTX3 ) increased by 45 % . P05231 and PTX3 appear to be valid biomarkers of the intradialytic inflammatory response . VC-related markers were in general not affected by the single HD session ; however , the observed correlation between acute changes of Q9GZV9 and phosphate during HD warrants further studies . Particle size of latex beads dictates IL-1β production mechanism . Macrophages ( Mϕ ) are well documented to produce IL-1β through various signaling pathways in response to small particles such as silica , asbestos and urea crystals , in the presence of lipopolysaccharide ( LPS ) . However , it has not been clear to what extent particle size affects the response . To investigate this point , we stimulated bone marrow-derived macrophages ( BMDM ) with size-defined latex beads ( LxB ) . Although both nano-sized ( 20 nm ) and micro-sized ( 1,000 nm ) LxB induced IL-1β production , only the nano-sized particles formed large intracellular vacuoles . In contrast , 100 nm LxB did not induce either of the responses . The same cellular responses were also observed in primary microglia cells . Although K(+) efflux and Q96P20 activation in BMDM were crucial in response to both 20 and 1,000 nm LxB , only IL-1β production by 20 nm LxB was sensitive to cathepsin B and Q99572 , a receptor for DB00171 . The response by 1,000 nm LxB relied on a robust production of reactive oxygen species ( ROS ) , since IL-1β production was remarkably reduced by ROS inhibitors such as diphenylene iodonium ( DPI ) and DB06151 ( Q9C000 ) . In contrast , IL-1β production by 20 nm LxB was augmented by Q9C000 and in BMDM deficient in thioredoxin-binding protein-2 ( P20226 -2 ) , a negative regulator of the ROS scavenger thioredoxin . These results suggest that the cells responded differently in their secretion of IL-1β depending on particle size , and that there is a range within which neither pathway works . DB00169 derivatives with adamantane or lactone ring side chains are cell type-selective vitamin D receptor modulators . The vitamin D receptor ( P11473 ) mediates the biological actions of 1,25-dihydroxyvitamin D(3) [ 1,25(OH)(2)D(3) ] , the active form of vitamin D , which regulates calcium homeostasis , immunity , cellular differentiation , and other physiological processes . We investigated the effects of three 1,25(OH)(2)D(3) derivatives on P11473 function . AD47 has an adamantane ring and LAC67a and LAC67b have lactone ring substituents at the side chain position . These vitamin D derivatives bind to P11473 but do not stabilize an active cofactor conformation . In a P11473 transfection assay , AD47 and LAC67b act as partial agonists and all three compounds inhibit P11473 activation by 1,25(OH)(2)D(3) . The derivatives enhanced the heterodimerization of P11473 with the retinoid X receptor , an effect unrelated to agonist/antagonist activity . AD47 and LAC67b weakly induced recruitment of the Q15788 cofactor to P11473 , and all three derivatives inhibited the recruitment of P52701 family cofactors to P11473 induced by 1,25(OH)(2)D(3) . It is noteworthy that AD47 induced Q15648 recruitment as effectively as 1,25(OH)(2)D(3) , whereas LAC67a and LAC67b were not effective . We examined the expression of endogenous P11473 target genes and the nuclear protein levels of P11473 and cofactors in several cell lines , including cells derived from intestine , bone , and monocytes , and found that the vitamin D(3) derivatives act as cell type-selective P11473 modulators . The data indicate that side chain modification is useful in the development of P11473 antagonists and tissue-selective modulators . Further elucidation of the molecular mechanisms of action of selective P11473 modulators will be essential for their clinical application . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . Synthesis and biological evaluation of 2- , 3- , and 4-acylaminocinnamyl-N-hydroxyamides as novel synthetic HDAC inhibitors . A new series of 2- , 3- , and 4-acylaminocinnamyl-N-hydroxyamides 1-3 have been prepared , and their anti-HDAC ( against maize Q92769 , HD1-B , and HD1-A enzymes ) activities have been assessed . Cinnamyl-hydroxyamides bearing acylamino substituents at the P06681 position of the benzene ring ( compounds 1a-g ) showed very low HDAC inhibiting activities , with IC(50) values in the high micromolar range . By shifting the same acylamino groups from P06681 to P01024 ( compounds 2a-g ) as well as C4 ( compounds 3a-f ) position of the benzene ring , a number of highly potent HDAC inhibitors have been obtained . In the anti- Q92769 assay 3c ( IC(50) = 11 nM ) was the most potent compound , being > 11600- , 4.5- , and 10-fold more potent than sodium valproate , DB02546 , and HC-toxin , respectively , and showing the same activity as trapoxin . HD1-B and HD1-A assays have been performed to screen the inhibitory action of 1-3 against mammalian class I ( HD1-B ) and class II ( HD1-A ) HDAC homologous enzymes . From the corresponding IC(50) data , a selectivity ratio has been calculated . In general , compounds 1-3 showed no or little selectivity towards the class II homologue HD1-A , the most selective being 2a with class II selectivity ratio = 4.3 . About the inhibitory potency , the 4-(2-naphthoylamino)cinnamyl-N-hydroxyamide 3f showed the highest inhibiting effect against the two enzymes ( IC(50-HD1-B) = 36 nM ; IC(50-HD1-A) = 42 nM ) . Selected 2 and 3 compounds will be evaluated to determine their antiproliferative and cyto differentiating activities on HL-60 cells . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation . P05231 , P01579 and P01375 production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 P01579 and P05231 by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 , P01579 and P01375 . These increases correlated with an increase in the numbers of P01730 + , CD8+ and CD25+ T cells in blood and P01730 + and CD25+ T cells in the liver perfusate , but not with CD8+ T cells in liver perfusate . Increased levels of P05231 , P01579 and P01375 were constitutively produced by liver-associated P01730 + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 + T cells secreted increasing levels of P01375 in a time-dependent manner following Con A injection , but P01375 production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 + T cells acting within the liver , at least in part through the secretion of P01375 , P01579 and P05231 . Synergy between vitamin D(3) and Toll-like receptor agonists regulates human dendritic cell response during maturation . Human dendritic cells ( DC ) can be differentiated from blood monocytes in the presence of GM- P04141 and P05112 and matured by lipopolysaccharide ( LPS ) . DB00169 inhibits the maturation of human DC measured by changes in surface expression of HLA-DR , P08571 , P25942 , P33681 , Q01151 , and P42081 . We here examine the function of vitamin D3 during DC maturation . One of the earliest changes to LPS-induced maturation was an increase in Q01151 expression . DB00169 inhibited the increase in expression of HLA-DR , P25942 , P33681 , Q01151 , and P42081 and the decrease in expression of P08571 , which was paralleled morphologically by vitamin D3-induced inhibition of dendritic cell differentiation . DB00169 acted in synergy with the TLR agonists LPS and peptidoglycan ( Q9UQ90 ) in inducing P05231 , P10145 , and P22301 , whereas vitamin D3 completely inhibited LPS-induced secretion of IL-12 . The synergy occurred at concentrations where neither vitamin D3 nor the TLR agonists alone induced measurable cytokine secretion . Both LPS and Q9UQ90 enhanced the level of the vitamin D3 receptor ( P11473 ) . Taken together , these data demonstrated that vitamin D3 and TLR agonists acted in synergy to alter secretion of cytokines from human DC in a direction that may provide an anti-inflammatory environment . P01148 antagonist DB00050 reduces prostate size and gene expression of proinflammatory cytokines and growth factors in a rat model of benign prostatic hyperplasia . BACKGROUND : Recent findings suggest that BPH has an inflammatory component . Clinical trials have documented that therapy with P01148 antagonist DB00050 causes a marked and prolonged improvement in LUTS in men with symptomatic BPH . We investigated the mechanism of action and effect of DB00050 in a rat model of BPH . METHODS : Adult male Wistar rats were used . BPH was induced in rats by subcutaneous injections of TE 2 mg/day for 4 weeks . Control animals received injections of corn oil . After induction of BPH , rats received depot DB00050 pamoate at the doses of 0.625 , 1.25 , and 12.5 mg/kg on days 1 and 22 and TE-control rats received vehicle injections . Whole prostates were weighed and processed for RNA and protein . Real-time RT-PCR assays for numerous inflammatory cytokines and growth factors were performed . Quantitative analyses of prostatic P01148 receptor , P01148 , androgen receptor ( AR ) and 5α-reductase 2 were done by real-time RT-PCR and immunoblotting ; serum DB02901 , LH , PSA , and DB01277 by immunoassays . RESULTS : mRNA levels for inflammatory cytokines IFN-γ , P08700 , P05112 , P05113 , P05231 , P10145 , P35225 , P40933 , and Q16552 and for growth factors P01133 , P09038 , FGF-7 , P55075 , Q92915 , TGF-β1 , and P15692 were significantly reduced by DB00050 0.625 mg/kg ( P < 0.05 ) . Prostate weights were also significantly lowered by any dose of DB00050 . CONCLUSIONS : This study suggests that DB00050 reduces various inflammatory cytokines and growth factors in rat prostate and , at doses which do not induce castration levels of testosterone , can lower prostate weights . Our findings shed light on the mechanism of action of P01148 antagonists in BPH . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Retinoic acid-induced modulation of P60568 mRNA production and P60568 receptor expression on T cells . BACKGROUND : Retinoic acid ( RA ) has important immune-modulating effects on both T and B cell function . Our laboratory has shown that RA can enhance in vitro polyclonal B cell immunoglobulin ( Ig ) response . Investigating cytokines known to affect B cell differentiation , we have recently shown that P05231 production is augmented by RA . In the present study we have examined the immune modulating effects of RA on P60568 mRNA , another important cytokine for B cell immunoglobulin production , the expression of P60568 receptors on T cells , and the RA nuclear receptors . METHODS : Purified T cells were obtained from adenoidal tissues , and incubated with RA ( 10(-7) M ) or DB01093 solvent/media control for 0 , 6-8 , and 24 h . Total mRNA was extracted from T cells , and using RT-PCR , changes in the production of P60568 and RA receptors ( RAR ) -alpha,beta,gamma mRNA were determined . The effects of RA on P60568 -alpha receptor expression was determined by flow cytometry on T cells . CONCLUSION : These studies suggest that RA can augment P60568 mRNA production by T cells with a possible paracrine effect on IL-2R-alpha expression . These changes appear to be mediated by P10276 . Thus , P60568 may be another important cytokine modulated by RA in the immune response . Q13507 -like protein and vitamin D receptor mediate DB00136 -induced Q5T124 influx in muscle cells . 1alpha,25-Dihydroxy-Vitamin-D3 ( 1alpha,25(OH)2- DB00169 ) stimulates in skeletal muscle cells Ca2+ release from inner stores and influx through both voltage-dependent and store-operated Ca2+ ( Q5T124 , CCE ) channels . We investigated the involvement of TRPC proteins and Vitamin D receptor ( P11473 ) in CCE induced by DB00136 in chick muscle cells . Two fragments were amplified by RT-PCR , exhibiting approximately 80 % sequence homology with mammalian Q13507 /6/7 . Northern and Western blots employing a Q13507 -probe and anti- Q13507 antibodies , respectively , confirmed endogenous expression of a Q13507 -like protein of 140 kDa . Spectrofluorimetric measurements in Fura-2 loaded cells showed reduced CCE and Mn2+ entry in response to either thapsigargin or DB00136 upon transfection with anti- Q13507 /6/7 antisense oligodeoxynucleotides ( ODNs ) . Transfection with anti- P11473 antisense ODNs diminished DB00136 -dependent Ca2+ and Mn2+ influx . Co-immunoprecipitation of Q13507 -like protein and P11473 under non-denaturating conditions was observed . We propose that endogenous Q13507 -like proteins and the P11473 participate in the modulation of CCE by DB00136 in muscle cells , which could be mediated by an interaction between these proteins . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . DB00169 down-regulates monocyte TLR expression and triggers hyporesponsiveness to pathogen-associated molecular patterns . Toll-like receptors ( TLR ) represent an ancient front-line defence system that enables the host organism to sense the presence of microbial components within minutes . As inducers of inflammation , TLR act as important triggers of distinct entities such as sepsis or autoimmune disease exacerbation . We report here that vitamin D3 [ 1alpha,25-dihydroxycholecalciferol , 1,25(OH)(2)D3 ] suppresses the expression of O60603 and O00206 protein and mRNA in human monocytes in a time- and dose-dependent fashion . Despite 1,25(OH)(2)D3-induced up-regulation of P08571 , challenge of human monocytes with either LPS or lipoteichoic acid resulted in impaired P01375 and procoagulatory tissue factor ( CD142 ) production , emphasizing the critical role of TLR in the induction of inflammation . Moreover , reduced TLR levels in 1,25(OH)(2)D3-treated phagocytes were accompanied by impaired NF-kappaB/RelA translocation to the nucleus and by reduced p38 and Q8NFH3 /44 ( extracellular signal-regulated kinase 1/2 ) phosphorylation upon TLR-ligand engagement . Both TLR down-regulation and P08571 up-regulation were substantially inhibited by the vitamin D receptor ( P11473 ) antagonist ZK 159222 , indicating that the immunomodulatory effect of 1,25(OH)(2)D3 on innate immunity receptors requires P11473 transcription factor activation . Our data provide strong evidence that 1,25(OH)(2)D3 primes monocytes to respond less effectively to bacterial cell wall components in a P11473 -dependent mechanism , most likely due to decreased levels of O60603 and O00206 . Attenuation of fever at near term : is interleukin-6- P40763 signalling altered ? Pregnant rats in late gestation show a reduced fever response after stimulation with lipopolysaccharide ( LPS ) . This can result from either an increased action of endogenous antipyretics or a reduction in the production or action of endogenous pyrogens . Nonpregnant rats given LPS release interleukin ( IL ) -6 , which causes nuclear translocation of the signal transducer and activator of transcription 3 ( P40763 ) in the vascular organ of the lamina terminalis ( OVLT ) , followed by a significant increase in core body temperature . The present study investigated whether the reduced fever response in near-term pregnant rats is associated with a reduced nuclear P40763 response . Rats at gestation day 15 ( Q99943 ) , gestation day 21 ( Q96NT5 , near term ) and at lactation day 5 ( Q15004 ) were injected with LPS ( 50 microg/kg , i.p. ) or vehicle . Only near-term pregnant rats responded with an attenuated body temperature during the fever response . Immunohistological analysis indicated no significant difference in nuclear P40763 in the OVLT of the different animal groups 2 h after LPS . Measurement of total and phosphorylated P40763 protein in the OVLT with semiquantitative western blot revealed no significant differences of this protein among these immune challenged animal groups . P05231 concentrations were also similar at Q99943 , Q96NT5 and Q15004 2 h after injection of LPS . These results lead to the conclusion that the attenuation of the fever response at near-term pregnancy is not associated with a reduced amount of nuclear P40763 in the OVLT , indicating a maintained P05231 - P40763 signalling pathway in the OVLT . Characterization of folate uptake by choroid plexus epithelial cells in a rat primary culture model . Reduced derivatives of folic acid ( folates ) play a critical role in the development , function and repair of the CNS . However , the molecular systems regulating folate uptake and homeostasis in the central nervous system remain incompletely defined . Choroid plexus epithelial cells express high levels of folate receptor alpha ( FRalpha ) suggesting that the choroid plays an important role in CNS folate trafficking and maintenance of P04141 folate levels . We have characterized 5-methyltetrahydrofolate ( 5-MTHF ) uptake and metabolism by primary rat choroid plexus epithelial cells in vitro . Two distinct processes are apparent ; one that is FRalpha dependent and one that is independent of the receptor . FRalpha binds 5-MTHF with high affinity and facilitates efficient uptake of 5-MTHF at low extracellular folate concentrations ; a lower affinity FRalpha independent system accounts for increased folate uptake at higher concentrations . Cellular metabolism of 5-MTHF depends on the route of folate entry into the cell . 5-MTHF taken up via a non-FRalpha -mediated process is rapidly metabolized to folylpolyglutamates , whereas 5-MTHF that accumulates via FRalpha remains non-metabolized , supporting the hypothesis that FRalpha may be part of a pathway for transcellular movement of the vitamin . The proton-coupled folate transporter , proton-coupled folate transporter ( Q96NT5 ) , mRNA was also shown to be expressed in choroid plexus epithelial cells . This is consistent with the role we have proposed for proton-coupled folate transporter in FRalpha-mediated transport as the mechanism of export of folates from the endocytic compartment containing FRalpha . The 1alpha,25-dihydroxy P11473 preferentially recruits the coactivator Q15788 during up-regulation of the osteocalcin gene . Binding of 1alpha,25-dihydroxy DB00169 to the C-terminal domain ( LBD ) of its receptor ( P11473 ) , induces a conformational change that enables interaction of P11473 with transcriptional coactivators such as the members of the P52701 / P12931 family or the DRIP ( Vitamin D interacting complex ) /Mediator complex . These interactions are critical for P11473 -mediated transcriptional enhancement of target genes . Recent reports indicate that nuclear receptors , including P11473 , interact with P52701 / P12931 members and the DRIP/Mediator complex in a sequential , cyclical , and mutually exclusive manner when bound to a target promoter , exhibiting also a high exchange rate . Here , we present an overview of how these coactivators are recruited to the bone-specific osteocalcin ( OC ) gene in response to short and long exposures to 1alpha,25-dihydroxy DB00169 . We find that in intact osteoblastic cells P11473 and Q15788 rapidly bind to the OC promoter in response to the ligand . This recruitment correlates with transcriptional enhancement of the OC gene and with increased histone acetylation at the OC promoter . In contrast , binding of the Q15648 subunit , which anchors the DRIP/Mediator complex to the P11473 , is detected at the OC promoter after several hours of incubation with 1alpha,25-dihydroxy DB00169 . Together , our results indicate that P11473 preferentially recruits Q15788 to enhance basal bone-specific OC gene transcription . We propose a model where specific protein-DNA and protein-protein interactions that occur within the context of the OC gene promoter in osteoblastic cells stabilize the preferential association of the P11473 - Q15788 complex . Replication of genetic association studies in aortic stenosis in adults . Only a handful of studies have attempted to unravel the genetic architecture of calcific aortic valve stenosis ( AS ) . The goal of this study was to validate genes previously associated with AS . Seven genes were assessed : P04114 , P02649 , P29279 , P22301 , PTH , P01137 , and P11473 . Each gene was tested for a comprehensive set of single-nucleotide polymorphisms ( SNPs ) . SNPs were genotyped in 457 patients who underwent surgical aortic valve replacement , and allele frequencies were compared to 3,294 controls . A missense mutation in the P04114 gene was significantly associated with AS ( rs1042031 , E4181K , p = 0.00001 ) . A second SNP located 5.6 kilobases downstream of the P04114 stop codon was also associated with the disease ( rs6725189 , p = 0.000013 ) . Six SNPs surrounding the P22301 locus were strongly associated with AS ( 0.02 > p > 6.2 × 10⁻¹¹ ) . The most compelling association for P22301 was found with a promoter polymorphism ( rs1800872 ) well known to regulate the production of the encoded anti-inflammatory cytokine . The frequency of the low-producing allele was greater in cases compared to controls ( 30 % vs 20 % , p = 6.2 × 10⁻¹¹ ) . SNPs in PTH , P01137 , and P11473 had nominal p values < 0.05 but did not resist Bonferroni correction . In conclusion , this study suggests that subjects carrying specific polymorphisms in the P22301 and P04114 genes are at higher risk for developing AS . Expression of vitamin D receptor and metabolizing enzymes in multiple sclerosis-affected brain tissue . Vitamin D deficiency has been implicated as a risk factor for multiple sclerosis ( MS ) , but how vitamin D metabolism affects MS pathophysiology is not understood . We studied the expression of vitamin D receptor ( P11473 ) and related enzymes , including 1,25(OH)(2)D-24-hydroxylase ( Q07973 ; Q07973 ) and 25(OH)D-1α-hydroxylase ( O15528 ) , in CNS tissues of 39 MS patients and 20 controls and in primary human glial cells in vitro . In control and MS normal-appearing white matter ( NAWM ) , nuclear P11473 immunostaining was observed in oligodendrocyte-like cells , human leukocyte antigen ( HLA ) -positive microglia , and glial fibrillary acidic protein-positive astrocytes . There was a 2-fold increase in P11473 transcripts in MS NAWM versus control white matter ( p = 0.03 ) . In chronic active MS lesions , HLA-positive microglia/macrophages showed nuclear P11473 staining ; astrocytes showed nuclear and cytoplasmic P11473 staining . Staining for Q07973 was restricted to astrocytes. P11473 and O15528 mRNA expressions were increased in active MS lesions versus NAWM ( p < 0.01 , p = 0.04 , respectively ) . In primary human astrocytes in vitro , the active form of vitamin D , 1,25(OH)(2)D(3) , induced upregulation of P11473 and Q07973 . P01375 and interferon-γ upregulated O15528 mRNA in primary human microglia and astrocytes . Increased P11473 expression in MS NAWM and inflammatory cytokine-induced amplified expression of P11473 and O15528 in chronic active MS lesions suggest increased sensitivity to vitamin D in NAWM and a possible endogenous role for vitamin D metabolism in the suppression of active MS lesions . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Regulation of the human P38936 (waf1/cip1) gene promoter via multiple binding sites for p53 and the vitamin D3 receptor . The main regulator of the human tumor suppresser gene P38936 (waf1/cip1) is the transcription factor p53 , but more recently it has been suggested to be a primary anti-proliferative target for the nuclear receptor P11473 in the presence of its ligand 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) . To identify P11473 responding regions , we analyzed 20 overlapping regions covering the first 7.1 kb of the P38936 (waf1/cip1) promoter in MCF-7 human breast cancer cells using chromatin immuno-precipitation assays ( ChIP ) with antibodies against p53 and P11473 . We confirmed two known p53 binding regions at approximate positions -1400 and -2300 and identified a novel site at position -4500 . In addition , we found three P11473 -associated promoter regions at positions -2300 , -4500 and -6900 , i.e. two regions showed binding for both p53 and P11473 . In silico screening and in vitro binding assays using recombinant and in vitro translated proteins identified five p53 binding sites within the three p53-positive promoter regions and also five DB00136 response elements within the three P11473 -positive regions . Reporter gene assays confirmed the expected responsiveness of the respective promoter regions to the p53 inducer 5-fluorouracil and DB00136 . Moreover , re-ChIP assays confirmed the functionality of the three DB00136 -reponsive promoter regions by monitoring simultaneous occupancy of P11473 with the co-activator proteins CBP , Q15788 and Q15648 . Taken together , we demonstrated that the human P38936 ( ( waf1/cip1 ) ) gene is a primary DB00136 -responding gene with at least three P11473 binding promoter regions , in two of which also p53 co-localizes . Apoptosis induction by 1alpha,25-dihydroxyvitamin D3 in prostate cancer . Calcitriol [ 1alpha,25-dihydroxyvitamin D3 ] is the natural ligand of the vitamin D receptor ( P11473 ) . Using cultured prostate cancer ( PC ) cell lines , LN-CaP and ALVA-31 , we studied the effects of 1alpha,25(OH)2- DB00169 ( VD3 ) on expression of several apoptosis-regulating proteins including : ( a ) Bcl-2 family proteins ( Bcl-2 , Bcl-X(L) , Mcl-1 , Bax , and Bak ) ; ( b ) the heat shock protein 70-binding protein BAG1L ; and ( c ) IAP family proteins ( P98170 , cIAP1 , and cIAP2 ) . VD3 induced decreases in levels of antiapoptotic proteins Bcl-2 , Bcl-X(L) , and Mcl-1 , BAG1L , P98170 , cIAP1 , and cIAP2 ( without altering proapoptotic Bax and Bak ) in association with increases in apoptosis . In contrast to P11473 -expressing LN-CaP and ALVA-31 cells , P11473 -deficient prostate cancer line Du-145 demonstrated no changes in apoptosis protein expression after treatment with VD3 . In sensitive PC cell lines , VD3 activates downstream effector protease , caspase-3 , and upstream initiator protease caspase-9 , the apical protease in the mitochondrial ( " intrinsic " ) pathway for apoptosis , but not caspase-8 , an initiator caspase linked to an alternative ( " extrinsic " ) apoptosis pathway triggered by cytokine receptors . VD3 induced declines in antiapoptotic proteins and also stimulated cytochrome c release from mitochondria by a caspase-independent mechanism . Moreover , apoptosis induction by VD3 was suppressed by overexpressing Bcl-2 , a known blocker of cytochrome c release , whereas the caspase-8 suppressor CrmA afforded little protection . Thus , VD3 is capable of inhibiting expression of multiple antiapoptotic proteins in P11473 -expressing prostate cancer cells , leading to activation of the mitochondrial pathway for apoptosis . The potential role of PD0332991 ( DB09073 ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 ) is an orally bioavailable , highly selective inhibitor of the P11802 /6-cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 /6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 /6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM . 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 . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . Expression of P20839 and P12268 after transplantation and initiation of immunosuppression . BACKGROUND : DB01024 ( DB00603 ) mediates immunosuppressive effects by inhibiting inosine monophosphate dehydrogenase ( IMPDH ) . Induction of IMPDH activity has been observed in whole blood and erythrocyte samples during immunosuppressive therapy . Information concerning the mechanisms for increased IMPDH activity is limited and the potential implications of induction have been debated . METHODS : Whole blood , P01730 + cell , and reticulocyte samples were collected from 30 renal transplant patients pre- and posttransplantation . The expressions of two IMPDH isoforms , type 1 and 2 , were analyzed by real-time reverse-transcription polymerase chain reaction and quantified using a housekeeping gene index . The IMPDH activity was determined by ultraviolet high-performance liquid chromatography . RESULTS : Transplantation and the initiation of immunosuppressive therapy was associated with increased P20839 ( 50-88 % , P < 0.0005 ) and decreased P12268 ( 42-56 % , P < 0.0005 ) expression . In P01730 + cells , however , P12268 increased ( 15 % , P=0.009 ) . These changes are probably related to glucocorticoid effects . Two weeks posttransplant , DB00603 -treated patients displayed elevated P20839 and 2 in reticulocytes , suggesting enzyme induction in these cells during prolonged DB00603 therapy . Patients with acute rejection during follow-up demonstrated higher P12268 expression in P01730 + cells pretransplant than nonrejecting patients ( median expression 1.26 vs. 0.87 respectively , P=0.017 ) . CONCLUSIONS : Knowledge of changes in P20839 and 2 expression after transplantation and initiation of immunosuppression is important considering the action of DB00603 on IMPDH and the potential for pharmacodynamic monitoring of DB00603 by measuring IMPDH activity . The expression of P12268 in P01730 + cells pretransplant may be an indicator of immune activation . Wogonin has multiple anti-cancer effects by regulating c-Myc/ Q13309 /Fbw7α and Q13547 / Q92769 pathways and inducing apoptosis in human lung adenocarcinoma cell line A549 . Wogonin is a plant monoflavonoid which has been reported to inhibit cell growth and/or induce apoptosis in various tumors . The present study examined the apoptosis-inducing activity and underlying mechanism of action of wogonin in A549 cells . The results showed that wogonin was a potent inhibitor of the viability of A549 cells . Apoptotic protein changes detected after exposure to wogonin included decreased P98170 and Mcl-1 expression , increased cleaved-PARP expression and increased release of O95831 and cytochrome C . Western blot analysis showed that the activity of c-Myc/Skp2 and Q13547 / Q92769 pathways , which play important roles in tumor progress , was decreased . Quantitative PCR identified increased levels of c-Myc mRNA and decreased levels of its protein . Protein levels of Fbw7α , GSK3β and Thr58-Myc , which are involved in c-Myc ubiquitin-dependent degradation , were also analyzed . After exposure to wogonin , Fbw7α and GSK3β expression decreased and Thr58-Myc expression increased . However , MG132 was unable to prevent c-Myc degradation . The present results suggest that wogonin has multiple anti-cancer effects associated with degradation of c-Myc , Q13309 , Q13547 and Q92769 . Its ability to induce apoptosis independently of Fbw7α suggests a possible use in drug-resistance cancer related to Fbw7 deficiency . Further studies are needed to determine which pathways are related to c-Myc and Fbw7α reversal and whether Thr58 phosphorylation of c-Myc is dependent on GSK3β . A case study of acenocoumarol sensitivity and genotype-phenotype discordancy explained by combinations of polymorphisms in Q9BQB6 and P11712 . To determine the cause of a genotype-phenotype discordancy for acenocoumarol sensitivity . Methods A patient , highly sensitive to acenocoumarol , and previously determined to carry only a single P11712 *3 allele , was genotyped for additional functionally defective alleles in the P11712 and Q9BQB6 genes . Family members were also analyzed to trace the pedigree . Results The acenocoumarol-sensitive patient was found to possess , in addition to P11712 *3 allele , a P11712 *11 allele and the Q9BQB6 AA diplotype which were all traced back through the parental lines . Conclusions DB01418 sensitivity in this subject is the consequence of inheritance of multiple functionally defective alleles in both the P11712 and Q9BQB6 genes . The study provides additional data in support of diminished P11712 activity due to the presence of the rare *11 allele . Retinoid receptors and acute promyelocytic leukaemia . While a great deal has been learned about APL over the last few years , many important questions remain unanswered . It has become clear that the P29590 / P10276 fusion protein is expressed in most cases of APL , and this protein presumably contributes to leukaemia initiation and/or progression . P29590 / P10276 appears to specifically block the further differentiation of myeloid progenitor cells , although the mechanism of its action is not known . It may inhibit the transcription of RAR- or P29590 -regulated genes , in which case expression must be restored in the presence of therapeutic RA concentrations . However , the possibility remains that P29590 / P10276 may have a novel function . In order to elucidate the molecular pathogenesis of APL , several important questions remain to be answered . These include whether P29590 is a transcription factor ; the identification of its target genes and response elements , and the role of P29590 / P10276 and RA in their regulation . Also whether the expression of P29590 / P10276 in bone marrow cells ( either by itself or in combination with other oncogenes ) alters their tumourigenicity or differentiation potential . It is also important to determine the function(s) of Q05516 and Q05516 / P10276 , and determine whether other APL patients with mutations involving P29590 or P10276 ( but not both ) respond to therapy with all-trans-RA . Finally , it is important both for the understanding of the molecular biology of APL and its therapy , to determine the effects of other regulatory factors involved in the control of myeloid cell differentiation such as granulocyte colony stimulation factor ( DB00099 ) and granulocyte macrophage colony stimulating factor ( GM- P04141 ) on APL cells in vitro and in vivo , both at presentation and in the RA-resistant patients in relapse . Role of transient receptor potential P01024 in P01375 -enhanced calcium influx in human airway myocytes . Previous studies have suggested that the proinflammatory cytokine , P01375 , contributes to airway hyperresponsivness by altering airway smooth muscle ( P17405 ) Ca(2+) responses to agonist stimulation . The present study examined the effects of P01375 on Ca(2+) influx pathways in cultured human P17405 cells ( HASMCs ) . Proteins encoded by the transient receptor potential ( TRP ) gene family function as channels through which receptor-operated and store-operated Ca(2+) entry ( SOCE ) occur . In the present study , the presence of P48995 , Q13507 , Q9UBN4 , Q9UL62 , and Q9Y210 mRNA and protein expression was confirmed in cultured HASMCs using RT-PCR and Western blot analysis . P01375 treatment significantly increased Q13507 mRNA and protein levels in HASMCs as well as SOCE . P01375 treatment also increased both the peak and plateau intracellular Ca(2+) concentration responses in HASMCs elicited by acetylcholine and bradykinin . The effects of P01375 treatment on SOCE and agonist-induced intracellular Ca(2+) concentration responses were attenuated using small interfering RNA transfection , which knocked down Q13507 expression . Thus , in inflammatory airway diseases , P01375 treatment may result in increased myocyte activation due to altered Ca(2+) influx pathways . These results suggest that Q13507 may be an important therapeutic target in inflammatory airway diseases such as asthma and chronic obstructive pulmonary disease . Oncogenic events associated with endometrial and ovarian cancers are rare in endometriosis . Endometriosis displays some features that resemble malignant processes , including invasive growth , resistance to apoptosis and distant implantation . The objective of this study was to investigate whether gene alterations that are frequent in endometrial and/or ovarian cancers contribute to the pathogenesis of endometriosis . Biopsies were obtained from ectopic endometriosis lesions from 23 patients with revised American Fertility Score stage 1 ( n= 1 ) , 2 ( n= 10 ) , 3 ( n= 11 ) or 4 ( n= 1 ) endometriosis . Six genes ( P25054 , CDKN2A , Q9ULZ3 , P10826 , Q9NS23 and P03372 ) were analyzed for promoter hypermethylation using methylation-specific melting curve analysis , and 9 genes ( P15056 , P01112 , P01111 , P35222 , P11802 , P22607 , P42336 , P04637 and P60484 ) were analyzed for mutations using denaturing gradient gel electrophoresis and direct sequencing . An oncogenic mutation in P01116 ( c.34G > T ; p.G12C ) was detected in a single lesion . No gene alterations were found in the remaining samples . Our data suggest that genetic and epigenetic events contributing to endometrial and ovarian cancers are rare in endometriosis . However , other proto-oncogenes and tumor suppressor genes should be tested for alterations in order to identify the molecular basis of the susceptibility of endometriosis to malignant transformation . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . Essential role of Ret for defining non-peptidergic nociceptor phenotypes and functions in the adult mouse . Transduction of pain following noxious stimuli is mediated by the activation of specialized ion channels and receptors expressed by nociceptive sensory neurons . A common early nociceptive sublineage expressing the nerve growth factor receptor TrkA diversifies into peptidergic and non-peptidergic nociceptors around birth . In this process , peptidergic neurons maintain TrkA expression , while non-peptidergic neurons downregulate TrkA and upregulate the common glial-derived neurotrophic factor family ligand receptor Ret and bind the isolectin B4 ( IB4 ) . Although Ret can have profound impacts on the molecular and physiological properties of nociceptive neurons , its role is not fully understood . Here we have deleted Ret in small- and medium-size sensory neurons , bypassing the early lethality of the full Ret knockout . We identify that Ret is expressed in two distinct populations of small-medium sized non-peptidergic neurons , an IB4(+) and an IB4(-) population . In these neurons , Ret is a critical regulator of several ion channels and receptors , including Nav1.8 , Nav1.9 , ASIC2a , P56373 , Q13507 , TrpM8 , TrpA1 , delta opioid receptor , MrgD , MrgA1 and MrgB4 . Ret-deficient mice fail to respond to mustard oil-induced neurogenic inflammation , have elevated basal responses and a failure to terminate injury-induced sensitization to cold stimuli , hypersensitivity to basal but not injury-induced mechanical stimuli , while heat sensation is largely intact . We propose that elevated pain responses could be contributed by Q9HC97 , which is dysregulated in adult Ret-deficient mice . Our results show that Ret is critical for expression of several molecular substrates participating in the detection and transduction of sensory stimuli , resulting in altered physiology following Ret deficiency . Novel vitamin D receptor ligands having a carboxyl group as an anchor to arginine 274 in the ligand-binding domain . DB00169 is metabolized into the hormonally active form , 1alpha,25-dihydroxyvitamin D3 ( 1 ) , via 25-hydroxyvitamin D3 ( 2 ) which is the most abundant circulating metabolite . Introduction of the 1alpha-hydroxyl group into 25-hydroxyvitamin D3 ( 2 ) to produce 1alpha,25-dihydroxyvitamin D3 ( 1 ) increases the P11473 binding affinity by approximately 1000-fold . The X-ray crystal structure of human P11473 in complex with 1alpha,25-dihydroxyvitamin D3 ( 1 ) shows that , together with DB00133 -237 , the 1alpha-hydroxyl group of 1alpha,25-dihydroxyvitamin D3 ( 1 ) makes hydrogen bonds with DB00125 -274 , single mutation of which results in impaired ligand recognition . In 2002 , lithocholic acid , which possesses a carboxyl group at position C24 , was demonstrated to be a weak P11473 ligand . We speculated that the carboxylic acid of lithocholic acid could be recognized by DB00125 -274 in the ligand-binding domain of P11473 . In view of the significance of DB00125 -274 to direct the 1alpha-hydroxyl group , as well as the results with lithocholic acid and its derivatives , we designed the P06681 modified analogues of 25-hydroxylvitamin D3 ( 2 ) having a carboxyl group , instead of the 1-hydroxyl group , for better electrostatic interaction to the guanidinium side-chain of arginine . TIF1delta , a novel P59665 -interacting member of the transcriptional intermediary factor 1 ( O15164 ) family expressed by elongating spermatids . O15164 ( transcriptional intermediary factor 1 ) proteins are encoded by an expanding family of developmental and physiological control genes that are conserved from flies to man . These proteins are characterized by an N-terminal RING-B box-coiled-coil ( Q9Y577 ) motif and a C-terminal P20941 finger/bromodomain unit , and have been implicated in epigenetic mechanisms of transcriptional repression involving histone modifiers and heterochromatin-binding proteins . We describe here the isolation and functional characterization of a fourth murine O15164 gene , TIF1delta . The predicted TIF1delta protein displays all the structural hallmarks of a bona fide O15164 family member and resembles the other TIF1s in that it can exert a deacetylase-dependent silencing effect when tethered to a promoter region . Moreover , like TIF1alpha and TIF1beta , TIF1delta can homodimerize and contains a PXVXL motif necessary and sufficient for P59665 ( heterochromatin protein 1 ) binding . Although TIF1alpha and TIF1beta also bind nuclear receptors and Kruppel-associated boxes specifically and respectively , TIF1delta appears to lack nuclear receptor- and Kruppel-associated box binding activity . Furthermore , TIF1delta is unique among the O15164 family proteins in that its expression is largely restricted to the testis and confined to haploid elongating spermatids , where it associates preferentially with P59665 isotype gamma ( HP1gamma ) and forms discrete foci dispersed within the centromeric chromocenter and the surrounding nucleoplasm . Collectively , these data are consistent with specific , nonredundant functions for the O15164 family members in vivo and suggest a role for TIF1delta in heterochromatin-mediated gene silencing during postmeiotic phases of spermatogenesis . P35367 occupancy in human brains after single oral doses of histamine H1 antagonists measured by positron emission tomography . 1. P35367 occupancy in the human brain was measured in 20 healthy young men by positron emission tomography ( PET ) using [ 11C ] -doxepin . 2 . (+)- DB01114 , a selective and classical antihistamine , occupied 76.8 +/- 4.2 % of the averaged values of available histamine H1 receptors in the frontal cortex after its administration in a single oral dose of 2 mg . Intravenous administration of 5 mg (+)-chlorpheniramine almost completely abolished the binding of [ 11C ] -doxepin to H1 receptors ( H1 receptor occupancy : 98.2 +/- 1.2 % ) . 3 . Terfenadine , a nonsedative antihistamine , occupied 17.2 +/- 14.2 % of the available H1 receptors in the human frontal cortex after its administration in a single oral dose of 60 mg . 4 . There was no correlation between H1 receptor occupancy by terfenadine and the plasma concentration of the active acid metabolite of terfenadine in each subject . 5 . PET data on human brain were essentially compatible with those on H1 receptor occupancy in guinea-pig brain determined by in vivo binding techniques , although for the same H1 receptor occupancy the dose was less in human subjects than in guinea-pigs . 6 . The PET studies demonstrated the usefulness of measuring H1 receptor occupancy with classical and second-generation antihistamines in human brain to estimate their unwanted side effects such as sedation and drowsiness quantitatively . [ Current Topics on Vitamin D. Evolution of animals and vitamin D ] . DB00169 is already found in the early evolution of life , but essentially as inactive products of the photochemical reaction of 7-dehydrocholesterol . The full vitamin D endocrine system characterized by the specific vitamin D transport protein ( DBP ) , specific vitamin D-metabolizing CYP P450 enzymes , active vitamin D metabolites , 1α,25 ( OH ) 2D3 , specific vitamin D nuclear receptor ( P11473 ) , and fibroblast growth factor 23 ( Q9GZV9 ) became essential for maintaining calcium and bone homeostasis in terrestrial animals cope with the challenging of higher gravity and calcium-poor environment . The present review describes the story about the evolution of animals and vitamin D . Evidence of an Epigenetic Modification in Cell-cycle Arrest Caused by the Use of Ultra-highly-diluted Gonolobus Condurango Extract . OBJECTIVES : Whether the ultra-highly-diluted remedies used in homeopathy can effectively bring about modulations of gene expressions through acetylation/deacetylation of histones has not been explored . Therefore , in this study , we pointedly checked if the homeopathically-diluted anti-cancer remedy Condurango 30C ( ethanolic extract of Gonolobus condurango diluted 10(-60) times ) was capable of arresting the cell cycles in cervical cancer cells HeLa by triggering an epigenetic modification through modulation of the activity of the key enzyme histone deacetylase 2 vis-a-vis the succussed alcohol ( placebo ) control . METHODS : We checked the activity of different signal proteins ( like P38936 (WAF) , p53 , Akt , P40763 ) related to deacetylation , cell growth and differentiation by western blotting and analyzed cell-cycle arrest , if any , by fluorescence activated cell sorting . After viability assays had been performed with Condurango 30C and with a placebo , the activities of histone de-acetylase ( HDAC ) enzymes 1 and 2 were measured colorimetrically . RESULTS : While Condurango 30C induced cytotoxicity in HeLa cells in vitro and reduced Q92769 activity quite strikingly , it apparently did not alter the Q13547 enzyme ; the placebo had no or negligible cytotoxicity against HeLa cells and could not alter either the HDAC 1 or 2 activity . Data on P38936 (WAF) , p53 , Akt , and P40763 activities and a cell-cycle analysis revealed a reduction in DNA synthesis and P55008 -phase cell-cycle arrest when Condurango 30C was used at a 2 % dose . CONCLUSION : Condurango 30C appeared to trigger key epigenetic events of gene modulation in effectively combating cancer cells , which the placebo was unable to do . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Discovery of [4-Amino-2-(1-methanesulfonylpiperidin-4-ylamino)pyrimidin-5-yl] ( 2,3-difluoro-6- methoxyphenyl ) methanone ( R547 ) , a potent and selective cyclin-dependent kinase inhibitor with significant in vivo antitumor activity . The cyclin-dependent kinases ( CDKs ) and their cyclin partners are key regulators of the cell cycle . Since deregulation of CDKs is found with high frequency in many human cancer cells , pharmacological inhibition of CDKs with small molecules has the potential to provide an effective strategy for the treatment of cancer . The 2,4-diamino-5-ketopyrimidines 6 reported here represent a novel class of potent and DB00171 -competitive inhibitors that selectively target the cyclin-dependent kinase family . This diaminopyrimidine core with a substituted 4-piperidine moiety on the P06681 -amino position and 2-methoxybenzoyl at the P01031 position has been identified as the critical structure responsible for the CDK inhibitory activity . Further optimization has led to a good number of analogues that show potent inhibitory activities against P06493 , P24941 , and P11802 but are inactive against a large panel of serine/threonine and tyrosine kinases ( K(i) > 10 microM ) . As one of these representative analogues , compound 39 ( R547 ) has the best CDK inhibitory activities ( K(i) = 0.001 , 0.003 , and 0.001 microM for P06493 , P24941 , and P11802 , respectively ) and excellent in vitro cellular potency , inhibiting the growth of various human tumor cell lines including an HCT116 cell line ( IC(50) = 0.08 microM ) . An X-ray crystal structure of 39 bound to P24941 has been determined in this study , revealing a binding mode that is consistent with our SAR . Compound 39 demonstrates significant in vivo efficacy in the HCT116 human colorectal tumor xenograft model in nude mice with up to 95 % tumor growth inhibition . On the basis of its superior overall profile , 39 was chosen for further evaluation and has progressed into Phase I clinical trial for the treatment of cancer . Vitamin D metabolism , mechanism of action , and clinical applications . DB00169 is made in the skin from 7-dehydrocholesterol under the influence of UV light . DB00153 ( ergocalciferol ) is derived from the plant sterol ergosterol . Vitamin D is metabolized first to 25 hydroxyvitamin D ( 25OHD ) , then to the hormonal form 1,25-dihydroxyvitamin D ( 1,25(OH)2D ) . Q6VVX0 is the most important 25-hydroxylase ; O15528 is the key 1-hydroxylase . Both 25OHD and 1,25(OH)2D are catabolized by Q07973 . 1,25(OH)2D is the ligand for the vitamin D receptor ( P11473 ) , a transcription factor , binding to sites in the DNA called vitamin D response elements ( VDREs ) . There are thousands of these binding sites regulating hundreds of genes in a cell-specific fashion . P11473 -regulated transcription is dependent on comodulators , the profile of which is also cell specific . Analogs of 1,25(OH)2D are being developed to target specific diseases with minimal side effects . This review will examine these different aspects of vitamin D metabolism , mechanism of action , and clinical application . Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin-1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA/uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 ) or interleukin-1 beta ( P01584 ) , are present in epidermal wounds . We have therefore tested P01584 and P01375 for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 and P01375 induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing . P12004 D3 interacts with vitamin D receptor and regulates its transcription activity . D-type cyclins are essential for the progression through the P55008 phase of the cell cycle . Besides serving as cell cycle regulators , D-type cyclins were recently reported to have transcription regulation functions . Here , we report that cyclin D3 is a new interacting partner of vitamin D receptor ( P11473 ) , a member of the superfamily of nuclear receptors for steroid hormones , thyroid hormone , and the fat-soluble vitamins A and D . The interaction was confirmed with methods of yeast two-hybrid system , in vitro binding analysis and in vivo co-immunoprecipitation . P12004 D3 interacted with P11473 in a ligand-independent manner , but treatment of the ligand , 1,25-dihydroxyvitamin D3 , strengthened the interaction . Confocal microscopy analysis showed that ligand-activated P11473 led to an accumulation of cyclin D3 in the nuclear region . P12004 D3 up-regulated transcriptional activity of P11473 and this effect was counteracted by overexpression of P11802 and Q00534 . These findings provide us a new clue to understand the transcription regulation functions of D-type cyclins . DB00169 suppresses the androgen-stimulated growth of mouse mammary carcinoma SC-3 cells by transcriptional repression of fibroblast growth factor 8 . Active metabolites of vitamin A and D are well known to act as growth inhibitors in hormone-related prostate and breast cancers . When various concentrations of 1alpha,25-dihydroxyvitamin D3 ( vitamin D3 ) , all-trans-retinoic acid ( DB00755 ) and 9-cis retinoic acid ( 9-cis RA ) were examined , the androgen-stimulated growth of mouse mammary carcinoma SC-3 cells was inhibited by vitamin D3 alone in a dose-dependent manner . A flow cytometer analysis showed that vitamin D3 leads SC-3 cells to relative P55008 -growth arrest after 72 h . Characterization of vitamin D3-responsive genes using an oligonucleotide microarray demonstrated that 220 genes were upregulated at more than threefold , and 84 genes were downregulated to less than one-third , compared with the testosterone-stimulated SC-3 cells . Neither cyclin-dependent kinase inhibitors ( CDKIs ) nor the antiapoptotic bcl-2 gene were induced in vitamin D3-responsive genes , with the exception of a slight induction of p15(INK4B) . Importantly , fgf8 was markedly repressed in response to vitamin D3 . The exogenous addition of P55075 canceled the growth suppression by vitamin D3 in SC-3 cells , suggesting that the repression of fgf8 is an indispensable step in vitamin D3-mediated growth inhibition . In reporter assays using the ARE-containing artificial construct and the natural androgen-regulated PSA promoter , co-transfection of the vitamin D receptor ( P11473 ) and androgen receptor ( AR ) suppressed AR-stimulated promoter activity . In addition , vitamin D3 also suppressed androgen-stimulated promoter activity in the stably transfected SC-3 cells . Moreover , P11473 repressed the core promoter activity of fgf8 in COS1 cells and in the SC-3 cells . All these findings strongly suggest that vitamin D3 serves as a negative regulator for both androgen-related and fgf8 transcriptions . Multiple pathways of apolipoprotein E signaling in primary neurons . P02649 is a genetic risk factor for Alzheimer 's disease , and the apoE protein is associated with beta-amyloid deposits in Alzheimer 's disease brain . We examined signaling pathways stimulated by apoE in primary neurons in culture . ApoE and an apoE-derived peptide activated several intracellular kinases , including prominently extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . P27361 /2 activation by apoE was blocked by an inhibitor of the low-density lipoprotein receptor family , the specific DB01221 glutamate receptor antagonist MK 801 and other calcium channel blockers . Activation of apoE receptors also induced tyrosine phosphorylation of Dab1 , an adaptor protein of apoE receptors , but experiments in Dab1 knockout neurons demonstrated that Dab1 was not necessary for P29323 activation . In contrast , apoE treatment of primary neurons decreased activation of c-Jun N-terminal kinase , a kinase that interacts with another apoE receptor adaptor protein , c-Jun N-terminal kinase-interacting protein . This change also depended on interactions with the low-density lipoprotein receptor family but was independent of calcium channels . c-Jun N-terminal kinase deactivation by apoE was blocked by gamma-secretase inhibitors and pertussis toxin . These results demonstrate that apoE affects several signaling cascades in neurons : increased disabled phosphorylation , activation of the P27361 /2 pathway ( dependent on calcium influx via the DB01221 receptor ) and inhibition of the P45983 /2 pathway ( dependent on gamma-secretase and G proteins ) . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . Mammalian Q99572 receptor pharmacology : comparison of recombinant mouse , rat and human Q99572 receptors . BACKGROUND AND PURPOSE : Acute activation of Q99572 receptors rapidly opens a non-selective cation channel . Sustained Q99572 receptor activation leads to the formation of cytolytic pores , mediated by downstream recruitment of hemichannels to the cell surface . Species- and single-nucleotide polymorphism-mediated differences in Q99572 receptor activation have been reported that complicate understanding of the physiological role of Q99572 receptors . Studies were conducted to determine pharmacological differences between human , rat and mouse Q99572 receptors . EXPERIMENTAL APPROACH : Receptor-mediated changes in calcium influx and Yo-Pro uptake were compared between recombinant mouse , rat and human Q99572 receptors . For mouse Q99572 receptors , wild-type ( BALB/c ) and a reported loss of function ( C57BL/6 ) Q99572 receptor were also compared . KEY RESULTS : BzATP [ 2,3-O-(4-benzoylbenzoyl)- DB00171 ] was more potent than DB00171 in stimulating calcium influx and Yo-Pro uptake at rat , human , BALB/c and C57BL/6 mouse Q99572 receptors . Two selective Q99572 receptor antagonists , A-740003 and A-438079 , potently blocked Q99572 receptor activation across mammalian species . Several reported P51575 receptor antagonists [ e.g. P59665 2159 ( 4- [ ( 4-formyl-5-hydroxy-6-methyl-3- [ (phosphonooxy)methyl } -2-pyridinyl ) azo ] -benzoic acid ) , PPNDS and NF279 ] blocked Q99572 receptors . NF279 fully blocked human Q99572 receptors , but only partially blocked BALB/c Q99572 receptors and was inactive at C57BL/6 Q99572 receptors . CONCLUSIONS AND IMPLICATIONS : These data provide new insights into Q99572 receptor antagonist pharmacology across mammalian species . Q99572 receptor pharmacology in a widely used knockout background mouse strain ( C57BL/6 ) was similar to wild-type mouse Q99572 receptors . Several structurally novel , selective and competitive Q99572 receptor antagonists show less species differences compared with earlier non-selective antagonists . P98170 regulates human interleukin-6 in umbilical vein endothelial cells via stimulation of the nuclear factor-kappaB and Q96HU1 kinase signaling pathways . X-linked inhibitior of apoptosis ( P98170 ) is known as a potent inhibitor of apoptosis , but more recently has been shown to also act as a modulator of the nuclear factor kB ( NF-kappaB ) signaling pathway . To investigate whether P98170 also affects other signalling pathways , we studied the transcriptional regulation of interleukin 6 ( P05231 ) , a gene that is strongly affected by P98170 , in more detail . The human P05231 gene contains transcription factor binding sites for activator protein 1 ( P05412 ) , enhancer binding protein beta ( C/EBP-beta ) and NF-kappaB . In reporter gene assays , mutation of these binding sites revealed the necessity of functional NF-kappaB and P05412 -sites for its ability to respond to P98170 . In contrast , P05231 promoter activity was slightly increased in the C/EBP deletion mutant . Pharmacologic inhibition of extracellular signal regulated kinase ( P29323 ) kinases ( Q02750 /2 ) as well as inhibition of the p38 signaling pathway both reduced P98170 -induced P05231 promoter activity . In conclusion , these results suggest that P98170 regulates P05231 transcription via NF-kappaB in cooperation with P05412 and C/EBP-beta .

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A review and expert opinion of the use of certolizumab for Crohn 's disease . INTRODUCTION : Crohn 's disease ( CD ) is a chronic , idiopathic , inflammatory bowel disease with no known cure . In those patients with moderate to severe disease , the result is often a clinically debilitating condition . In the last decade , one of the most significant developments in therapy has been a class of biological agents that neutralize TNFa . DB08904 ( CZP ) is the most recently FDA approved anti- P01375 agent for the induction and maintenance of moderate to severely active Crohn 's disease . AREAS COVERED : The currently available evidence regarding the use of CZP in CD , the expected efficacy and possible adverse events associated with this population . EXPERT OPINION : CZP is a TNFa inhibitor that is a safe and effective agent for treatment of CD . It has several unique features which make it useful in patients with moderate to severe disease . Cordycepin suppresses P01375 -α-induced NF-κB activation by reducing p65 transcriptional activity , inhibiting IκBα phosphorylation , and blocking IKKγ ubiquitination . Cordycepin is reported to participate in multiple pharmacological activities including anti-tumor and anti-inflammation , and is involved in the regulation of NF-κB signaling pathway . However , the detailed molecular mechanism of cordycepin in suppression of NF-κB signaling pathway remains ambiguous . In this study , we first analyzed the effect of cordycepin on NF-κB activity in P29320 -293T cells , and found that cordycepin resulted in a dose-dependent reduction in P01375 -α-induced NF-κB activation . Although cordycepin did not block P01375 -α-induced nuclear translocation of p65 , high concentration of cordycepin reduced the DNA-binding and transcriptional activities of NF-κB . Moreover , cordycepin also inhibited IκBα phosphorylation so as to suppress the degradation of IκBα . Further investigation revealed that cordycepin suppressed IKKs-mediated NF-κB activation and inhibited the ubiquitination of IKKγ . In conclusion , cordycepin effectively inhibits NF-κB signaling through suppressing the activities of NF-κB , IκB and IKK . Thus , cordycepin may provide some potential therapeutic application in inflammation-associated disorders and cancer . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . Pulmonary infectious complications of tumor necrosis factor blockade . The understanding of the infection risks posed by tumor necrosis factor ( P01375 ) antagonists has continued to evolve in the 10 years since these drugs first were introduced . Recent prospective studies have confirmed the risk of tuberculosis ( TB ) reactivation posed by P01375 antibodies to be several fold greater than soluble P01375 receptor . DB08904 , a monovalent anti- P01375 Fab ' fragment , appears to share this risk , despite its lack of Fc and its inability to cross-link transmembrane P01375 or activate complement . Two-step ( boosted ) tuberculin skin test screening and initiation of treatment for latent TB infection can greatly reduce the TB risk of anti- P01375 treatment in western countries . Current recommendations for withdrawal of anti- P01375 therapy when TB is diagnosed place patients at risk for paradoxical worsening due to recovery of P01375 -dependent inflammation . Further research is needed to determine how best to prevent and manage their infectious complications and to determine their potential adjunctive therapeutic role in chronic infectious diseases . DB08904 for the treatment of psoriatic arthritis . DB08904 ( CZP ) is a P01375 -α inhibitor approved for the treatment of psoriatic arthritis in 38 countries , including many European countries and the USA . It is a pegylated humanized anti- P01375 -α antigen-binding fragment , administered subcutaneously . As other P01375 -α antibodies , CZP binds to and neutralizes both soluble and membrane P01375 -α . In contrast to whole antibodies and etanercept , CZP does not activate antibody-dependent cell-mediated cytotoxicity or complement-dependent cytotoxicity , as it does not have an Fc piece . CZP showed efficacy in improving skin scores and patient reported outcomes in a Phase II study of 176 adults with moderate-to-severe plaque psoriasis . In a Phase III study of CZP in 409 psoriatic arthritis patients , CZP treatment resulted in improvements in peripheral arthritis , as well as dactylitis , enthesitis , nail disease and quality of life . The safety profile of CZP appears to be similar to that of other P01375 -α inhibitor . How future tumor necrosis factor antagonists and other compounds will meet the remaining challenges in Crohn 's disease . The chimeric monoclonal antibody to tumor necrosis factor ( P01375 ) , infliximab , is an effective therapy for Crohn 's disease . However , the formation of human anti-chimeric antibodies to infliximab ( immunogenicity ) can lead to loss of efficacy as well as acute infusion reactions and delayed hypersensitivity reactions . The fully human monoclonal antibody adalimumab and the pegylated humanized monoclonal antibody fragment DB08904 are biologic therapies against P01375 that might be effective for Crohn 's disease and less immunogenic than infliximab . Other potential alternatives to infliximab for Crohn 's disease include the humanized anti-adhesion molecule antibodies natalizumab and DB05802 , the humanized anti-interleukin 12 antibody ABT-874 , the humanized anti-interferon g antibody fontolizumab , the humanized anti-interleukin 6 receptor antibody DB06273 , and human recombinant granulocyte macrophage colony stimulating factor ( sargramostim ) . Some , or all , of these therapies will likely represent important treatments for Crohn 's disease in the future . Anti- P01375 therapy in Crohn 's disease . Anti-tumour necrosis factor ( P01375 ) strategies , the most studied of biological therapies , include chimeric monoclonal ( infliximab ) , humanized monoclonal ( CDP571 and the PEGylated DB08904 ) and fully human monoclonal ( adalimumab ) antibodies , p75 fusion protein ( etanercept ) , p55 soluble receptor ( onercept ) and small molecules such as MAPkinase inhibitors . The principal use of infliximab is in treating active Crohn 's disease patients not responding to or intolerant of conventional therapies . DB00065 is steroid sparing . The development of antibodies against infliximab is associated with an increased risk of infusion reactions , and a reduced duration of response to treatment , and concomitant immunosuppressive therapy reduces the immunogenic response . The demonstration of the efficacy of maintenance therapy every 8 weeks with infliximab in the randomised , controlled , ACCENT I trial opened up the strategy for regular maintenance . In patients who have failed therapy with cortocosteroids and immunosuppressive therapy and are poor surgical candidates , and patients with fistulizing disease , where infliximab therapy is chosen , regular maintenance therapy with infliximab is likely to be required . On the other hand , patients with severely active , steroid-refractory disease in whom immunosuppressive therapy and infliximab are initiated together , may respond adequately and be continued on long-term immunosuppressive therapy alone . In ulcerative colitis the role of infliximab remains uncertain . Simultaneous inhibition of epidermal growth factor receptor ( P00533 ) signaling and enhanced activation of tumor necrosis factor-related apoptosis-inducing ligand ( P50591 ) receptor-mediated apoptosis induction by an scFv:sTRAIL fusion protein with specificity for human P00533 . P00533 ( P00533 ) signaling inhibition by monoclonal antibodies and P00533 -specific tyrosine kinase inhibitors has shown clinical efficacy in cancer by restoring susceptibility of tumor cells to therapeutic apoptosis induction . P01375 -related apoptosis-inducing ligand ( P50591 ) is a promising anti-cancer agent with tumor-selective apoptotic activity . Here we present a novel approach that combines P00533 -signaling inhibition with target cell-restricted apoptosis induction using a P50591 fusion protein with engineered specificity for P00533 . This fusion protein , scFv425:sTRAIL , comprises the P00533 -blocking antibody fragment scFv425 genetically fused to soluble P50591 ( sTRAIL ) . Treatment with scFv425:sTRAIL resulted in the specific accretion to the cell surface of P00533 -positive cells only . P00533 -specific binding rapidly induced a dephosphorylation of P00533 and down-stream mitogenic signaling , which was accompanied by cFLIP(L) down-regulation and Bad dephosphorylation . P00533 -specific binding converted soluble scFv425:sTRAIL into a membrane-bound form of P50591 that cross-linked agonistic P50591 receptors in a paracrine manner , resulting in potent apoptosis induction in a series of P00533 -positive tumor cell lines . Co-treatment of P00533 -positive tumor cells with the P00533 -tyrosine kinase inhibitor DB00317 resulted in a potent synergistic pro-apoptotic effect , caused by the specific down-regulation of O15519 . Furthermore , in mixed culture experiments binding (L)of scFv425:sTRAIL to P00533 -positive target cells conveyed a potent apoptotic effect toward P00533 -negative bystander tumor cells . The favorable characteristics of scFv425:sTRAIL , alone and in combination with DB00317 , as well as its potent anti-tumor bystander activity indicate its potential value for treatment of P00533 -expressing cancers . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . 8-OH-DPAT ( P08908 agonist ) Attenuates 6-Hydroxy- dopamine-induced catalepsy and Modulates Inflammatory Cytokines in Rats . OBJECTIVE(S) : Neuroinflammation in Parkinson disease ( PD ) is associated with glial cells activation and production of different inflammatory cytokines . In this study , we investigated the effect of chronic administration of 8-OH-DPAT on 6-OHDA-induced catalepsy and levels of inflammatory cytokines in cerebrospinal fluid ( P04141 ) . MATERIALS AND METHODS : Catalepsy was induced by unilateral infusion of 6-OHDA ( 8 μg/2 μl/rat ) into the central region of the sabstantia nigra pars compacta ( SNc ) being assessed by the bar-test , 5 , 60 , 120 and 180 min after intraperitoneal ( IP ) administration of 8-OH-DPAT ( P08908 receptor agonist ; 0.25 , 0.5 and 1mg/kg , IP for 10 days ) . P04141 samples were collected on the tenth day of 8-OH-DPAT administration and analyzed by ELISA method to measure levels of P01375 -α , IL-1β and P05231 . RESULTS : Chronic injection of 8-OH-DPAT decreased catalepsy in a dose dependent manner when compared with the control group . The most anti-cataleptic effect was observed at the dose of 1 mg/kg of 8-OH-DPAT . Levels of P01375 -α in P04141 increased three weeks after 6-OHDA injection while there was a significant decrease in P01375 -α level of parkinsonian animals treated with 8-OH-DPAT ( 1 mg/kg , IP for 10 days ) . IL-1β and P05231 decreased and increased in parkinsonian rats and in 8-OH-DPAT-treated parkinsonian rats , respectively . CONCLUSION : Our study indicated that chronic administration of 8-OH-DPAT improves catalepsy in 6-OHDA-induced animal model of PD and restores central concentration of inflammatory cytokines to the basal levels . P08908 receptor agonists can be suggested as potential adjuvant therapy in PD by modulation of cerebral inflammatory cytokines . DB08904 for the treatment of rheumatoid arthritis . INTRODUCTION : Improved understanding of the pathogenesis of rheumatoid arthritis ( RA ) , and subsequent development of targeted therapies , have greatly advanced the management of this chronic inflammatory disease . The aim of treatment is a state of clinical remission . DB08904 ( CZP ) is a novel pegylated P01375 alpha inhibitor ( TNFi ) therapy and is the focus of this review . AREAS COVERED : CZP is different from other TNFi as it contains a polyethylene glycol ( PEG ) moiety , and lacks the constant fragment ( Fc ) of immunoglobulin ; therefore it does not activate complement . In this review in addition to clinical efficacy of CZP , effects on radiographic and patient-reported outcomes , are discussed . Adverse event data from clinical trials and extension studies are also reviewed . EXPERT OPINION : The addition of novel TNFi therapies to treat RA is welcomed . As well as displaying clinical efficacy , there is evidence to suggest that CZP has unique characteristics , including reduced transfer across the placenta and reduced frequency of injection site reactions . Furthermore , randomised controlled trials ( RCTs ) of CZP have demonstrated rapid improvements in workplace and home productivity in patients contributing to reducing the significant socioeconomic burden of RA . Recommendations for the treatment of Crohn 's disease with tumor necrosis factor antagonists : an expert consensus report . BACKGROUND : Symptom relief is the traditional treatment goal in Crohn 's disease ( CD ) . New goals including mucosal healing and bowel preservation are now achievable with tumor necrosis factor ( P01375 ) antagonists . DB00065 and adalimumab are approved as second-line treatments for severe , active CD . DB08904 is approved only in the U.S. and Switzerland as second-line treatment for moderate-to-severe , active CD . Data from trials of infliximab suggest that high-risk patients and patients with active inflammation ( CRP elevation and/or ileocolonic ulcers ) may benefit from earlier use of this drug . METHODS : A Delphi survey was used to obtain consensus on issues surrounding bowel preservation and use of P01375 antagonists . At the time of this survey , infliximab was the only P01375 antagonist approved for the treatment of CD in Europe , Canada , and Australia . An expert panel of 12 gastroenterologists with substantial clinical experience using infliximab in clinical practice and trials in these areas participated . RESULTS : The experts agreed that bowel preservation and mucosal healing are relevant and achievable goals , and form a rationale for using P01375 antagonists in CD patients . Control of inflammation and induction of mucosal healing were considered essential for bowel preservation . Consensus areas : 1 ) mucosal healing is predictive of improved long-term disease course and increases the likelihood of steroid-free remission ; 2 ) infliximab induces sustained mucosal healing , promotes bowel preservation , and reduces hospitalizations and surgeries ; 3 ) benefits of infliximab in relation to mucosal healing , bowel preservation , and clinical remission increase when therapy is initiated earlier . CONCLUSIONS : Treatment with P01375 antagonists helps preserve the bowel in CD patients . Targeting nanomedicines in the treatment of Crohn 's disease : focus on certolizumab pegol ( DB08904 ) . A variety of targets for therapeutic intervention are based upon advances in understanding of the immunopathogenesis of Crohn 's disease . Crohn 's disease is initiated by an innate immune response , which eventuates in a T-cell driven process , characterized by a T-helper cell 1 type cytokine profile . Several new treatments now focus on suppressing T-cell differentiation or T-cell inflammation . Since inflammatory bowel disease ( Q9UKU7 ) represents a state of dysregulated inflammation , drugs that augment the anti-inflammatory response have the potential to downregulate inflammation and thereby hopefully modify the disease . Tumour necrosis factor ( P01375 ) is a major target of research and clinical investigation . P01375 has proinflammatory effects in the intestinal mucosa and is a pivotal cytokine in the inflammatory cascade . DB08904 ( DB08904 ) is a PEGylated , Fab ' fragment of a humanized anti- P01375 monoclonal antibody . PEGylation increases the half-life , reduces the requirement for frequent dosing , and possibly reduces antigenicity as well . Certolizumab has been shown in Phase III trials to achieve and maintain clinical response and remission in Crohn 's disease patients . It improves the quality of life . DB08904 will be indicated for moderately to severely active Crohn 's disease , but it is not yet licensed in Europe or the US . It is not possible to construct an algorithm for treatment , but when compared with infliximab the two principal advantages are likely to be lower immunogenicity ( as shown by anti-drug antibodies , absence of infusion reactions , and low rate of antinuclear antibodies ) , and a subcutaneous route of administration . These two factors may be sufficient to promote it up the pecking order of anti- P01375 agents . DB08904 in axial spondyloarthritis . The axial spondyloarthritis ( SpA ) classification criteria cover both patients with ankylosing spondylitis and non-radiographic axial SpA . After failure of NSAIDs P01375 -α-inhibitors ( P01375 -blockers ) can be given to patients with active axial SpA . Until recently , the P01375 -blockers infliximab , adalimumab , etanercept and DB06674 are labeled for the treatment of active ankylosing spondylitis while for active nr-axSpA only adalimumab has been approved in Europe . The P01375 -blocker certolizumab pegol has recently been evaluated in the RAPID-axSpA trial which is the first placebo-controlled randomized-controlled trial in the entire group of axial SpA . An elevated P02741 and/ or evidence of bone marrow edema on Q9BWK5 of the sacroiliac joints were required for inclusion in RAPID-axSpA , and patients could have been preexposed to P01375 -blockers . The interesting data of this important trial in the context of the emerging therapeutic field of non-radiographic axial SpA therapy is discussed in this review . DB08904 in the treatment of Crohn 's disease . INTRODUCTION : The introduction of antibodies directed against tumour necrosis factor ( anti- P01375 ) has dramatically changed the concept of treating patients with Crohn 's disease ( CD ) . Unfortunately , the long-term efficacy of anti- P01375 agents may be hampered by immunogenicity . The availability of more anti- P01375 agents in the therapeutic armamentarium would therefore be of great benefit in patients loosing response to another anti- P01375 . In this review , the authors will focus on the efficacy of certolizumab pegol ( CZP ) . AREAS COVERED : A literature search to January 2013 was performed to identify all trials studying CZP in patients with CD . The authors first focused on the mechanism of action of CZP . Second , they evaluated the efficacy of an induction and maintenance therapy with CZP and the impact of CZP on mucosal healing and fistula closure . Next , they explored the influence of previous anti- P01375 exposure and baseline P02741 levels . Finally , they analysed the safety data on CZP , including the development of antibodies against CZP and the use of CZP during pregnancy . EXPERT OPINION : Based on the provided literature , CZP could be a good alternative in patients with moderate-to-severe CD failing another anti- P01375 agent . More data are required to conclude on its impact on the long-term outcome of CD . Targeting tumor necrosis factor alpha in psoriasis and psoriatic arthritis . BACKGROUND : Psoriasis is an immune-mediated chronic inflammatory disease triggered and maintained by inflammatory mediators , including P01375 . OBJECTIVE/METHODS : To summarize the role of anti- P01375 agents psoriasis therapy , focusing on the mechanisms and biological pathways involved , by reviewing relevant literature . RESULTS/CONCLUSIONS : The three P01375 antagonists currently available ( etanercept , infliximab and adalimumab ) are effective in the therapy of psoriasis and psoriatic arthritis . DB08904 and DB06674 are P01375 inhibitors not approved for therapy of psoriasis yet . In addition to neutralizing soluble P01375 , P01375 blockers bind to membrane P01375 and change the behavior of P01375 -expressing cells , resulting in hastened cell cycle arrest and apoptosis , and suppression of cytokine production . P01375 blockers may also affect adaptive immune responses by reducing T helper cell (Th)1 and Th17 responses , and favoring the development of T-regulatory cells . P01375 antagonists can regulate differentiation and activation of osteoclasts , thus reducing bone destruction in psoriatic arthritis . Anti- P01375 agents differ in their pharmacokinetics and pharmacodinamic properties , which is reflected in their therapeutic and safety profiles . The safety of P01375 antagonists has been established , and patient selection and monitoring allow risk minimization . DB08904 for the treatment of Crohn 's disease . DB08904 is a polyethylene glycolated FAb ' fragment of a humanized anti- P01375 monoclonal antibody . This pegylated molecule binds with circulating P01375 and forms an inactive complex that is then eliminated from the body . The drug has been shown to be better than placebo in the treatment of Crohn 's disease and maintaining a clinical response in adult patients with moderate-to-severe active disease who have had an inadequate response to conventional therapy , and the treatment of adults with moderately to severely active rheumatoid arthritis . Comparative trials with an active control group are lacking . The most common adverse reactions include abdominal pain , diarrhea , injection site reactions and infection . All necessary live and attenuated vaccines should be given prior to the initiation of certolizumab pegol therapy , patients should be evaluated for TB risk factors and tested for latent TB prior to initiating therapy , and the initiation of therapy should be avoided if the patient has an active infection . Concomitant use with anakinra is not recommended because of the increased risk of serious infections and neutropenia . Therapy should be discontinued if the patient develops a serious infection during therapy . The use of P01375 blocking agents in rheumatoid arthritis : an update . P01375 has been found to play a pivotal role in the pathogenic mechanisms of rheumatoid arthritis ( RA ) . Drugs targeting P01375 have been developed to neutralise the deleterious effects of this inflammatory cytokine . There are , at present , three drugs available for the treatment of RA patients with active disease who are refractory to conventional treatments including methotrexate : 2 monoclonal antibodies , infliximab and adalimumab , and a fusion protein with p75 receptors , etanercept . These three agents have proved to be effective and safe in large placebo-controlled trials enrolling patients with established or early disease and showed effectiveness in controlling signs and symptoms of the disease , improving quality of life and in slowing and even arresting the progression of radiographic damage . With the long-term surveillance of these drugs were described serious adverse events , particularly infections such as tuberculosis , especially with infliximab . The risk for malignancies under P01375 antagonists , especially lymphoma , remains controversial . Specific recommendations are given by international experts for selecting and monitoring RA patients with P01375 antagonists . Other drugs targeting P01375 such as PEGylated molecules ( DB08904 or certolizumab ) are in development . These new biological therapies blocking P01375 undoubtedly constitute a considerable advancement in the management of RA , but careful evaluation at the initiation of the treatment and long-term surveillance of the patients receiving such drugs remains necessary . Prefilled certolizumab pegol ( Cimzia(®) ) syringes for self-use in the treatment of rheumatoid arthritis . A new anti-tumor necrosis factor alpha ( P01375 -α ) inhibitor with a novel mechanism of action has entered phase 3 trials in rheumatoid arthritis ( RA ) . DB08904 ( Cimzia(®) ) is a humanized Fab ' antibody fragment against P01375 -α with a polyethylene glycol tail that prevents complement-dependent and antibody-dependent cell-mediated cytotoxicity or apoptosis . Four randomized clinical trials have been published so far . Reported results are similar to those published in previous studies with other P01375 -α inhibitors , with ACR20 , ACR50 , and ACR70 responses of around 60 % , 40 % , and 20 % , respectively , when combined with methotrexate and slightly lower when used as monotherapy . Safety was shown to be similar to that seen with P01375 -α blockers and some cases of tuberculosis were seen in the trials , stressing the importance of a complete screening in these patients . Although we still need effectiveness and safety data in larger numbers of patients and longer follow-up , this new P01375 inhibitor is a welcome addition to our current armamentarium for the treatment of RA . DB08904 in rheumatoid arthritis : current update . INTRODUCTION : The development of P01375 -α inhibitors ( P01375 -is ) represents a major advancement in the treatment of rheumatoid arthritis ( RA ) . Currently , there are five agents licensed for moderate-to-severely active RA . DB08904 ( CZP ) is a novel PEGylated , constant fragment-free P01375 -i therapy , which is the focus of this review . AREAS COVERED : Data from Phase III randomised controlled trials in terms of clinical efficacy , radiographic progression , patient-reported outcomes and safety profile are reviewed . These include long-term data from open-label extension studies . EXPERT OPINION : The advantages of CZP include rapid reduction of disease activity , low rates of injection-site reaction and may be safe for use in pregnancy . The long-term data strengthen the position of CZP for use either as monotherapy or preferably in combination with disease modifying anti-rheumatic drugs ( DMARDs ) , in moderate-to-severely active RA , comparable to other P01375 -is . Notably , prolonged CZP exposure is not associated with increased risk of severe infection compared to general population , contrasting with preliminary analysis of short-term data . Over the next few years , evidence will be available on the use of CZP in combination with methotrexate for remission induction in DMARD-naïve patients , biomarkers and the development and licensing of P01375 -i biosimilars . P01375 and its inhibitors in cancer . P01375 ( P01375 ) -alpha is implicated in the same time in apoptosis and in cell proliferation . P01375 not only acts as pro-inflammatory cytokine conducing to wide spectrum of human diseases including inflammatory diseases , but can also induce tumor development . The molecular mechanisms of P01375 functions have been intensively investigated . In this review we covered P01375 , the molecule , its signaling pathway , and its therapeutic functions . We provide a particular insight in its paradoxical role in tumor promotion and in its use as anti-tumor agent . This review considers also the recent findings regarding P01375 inhibitors , their pharmacokinetics , and their pharmacodynamics . Six P01375 inhibitors have been considered here : DB00065 , DB00051 , Golimumab , DB08904 , CDP571 , DB00005 , and Thalidomide . We discussed the clinical relevance of their functions in treatment of several diseases such as advanced inflammatory rheumatic and bowel disease , with a focus in cancer treatment . Targeting P01375 by these drugs has many side effects like malignancies development , and the long-term sequels are not very well explored . Their efficacy and their safety were discussed , underscoring the necessity of close patients monitoring and of their caution use . P01375 as a therapeutic target in inflammatory diseases , ischemia-reperfusion injury and trauma . P01375 -alpha ( P01375 ) is a central regulator of inflammation , and P01375 antagonists may be effective in treating inflammatory disorders in which P01375 plays an important pathogenetic role . Recombinant or modified proteins are an emerging class of therapeutic agents . To date , several recombinant or modified proteins which acts as P01375 antagonists have been disclosed . In particular , antibodies that bind to and neutralise P01375 have been sought as a means to inhibit P01375 activity . Inhibition of P01375 has proven to be an effective therapy for patients with rheumatoid arthritis and other forms of inflammatory disease including psoriasis , psoriatic arthritis , and ankylosing spondylitis , inflammatory bowel disease . Additionally , the efficacy of preventing septic shock and AIDS has been questioned as a result of recent research . The currently available therapies include a soluble p75 P01375 receptor:Fc construct , etanercept , a chimeric monoclonal antibody , infliximab , and a fully human monoclonal antibody , adalimumab . DB08904 is a novel P01375 inhibitor which is an antigen-binding domain of a humanized P01375 antibody coupled to polyethylene glycol ( PEG ) to increase half-life , and thus is Fc-domain-free . In this review , we discuss briefly the present understanding of P01375 -mediated biology and the current therapies in clinical use , and focus on some of the new therapeutic approaches with small-molecule inhibitors . Moreover , we examine recent reports providing important insights into the understanding of efficacy of thalidomide and its analogs , as P01375 activity inhibitories , especially in therapies of several inflammatory diseases within the nervous system . Randomised clinical trial : improvement in health outcomes with certolizumab pegol in patients with active Crohn 's disease with prior loss of response to infliximab . BACKGROUND : Crohn 's disease ( CD ) is associated with impaired health-related quality of life ( HRQoL ) . DB08904 , administered either every 2 weeks ( q2w ) or q4w , maintains efficacy in patients previously failing on the anti- P01375 agent infliximab ( WELCOME study ) . AIM : To investigate the impact of certolizumab pegol administered q2w and q4w on work productivity and HRQoL in the WELCOME study . METHODS : Patients with loss of response to infliximab received open-label certolizumab pegol induction and were randomised to receive double-blind maintenance treatment with certolizumab pegol 400 mg either q4w or q2w through week 24 , with a final evaluation at week 26 . Work productivity and HRQoL were assessed using the Work Productivity and Activity Impairment:CD questionnaire and Inflammatory Bowel Disease Questionnaire respectively . RESULTS : Baseline HRQoL burden was representative of moderately to severely active CD . HRQoL , daily activity and work productivity improved in both treatment groups as early as week 6 and were maintained through week 26 . Treatment benefits to HRQoL , daily activity and work productivity were similar between the certolizumab pegol q2w vs. q4w groups . CONCLUSIONS : DB08904 therapy results in meaningful improvements in work productivity , daily activities and HRQoL in patients with active CD who previously responded to but either lost response or could not tolerate infliximab ( ClinicalTrials.gov number : NCT00308581 ) . DB08904 : a new biologic targeting rheumatoid arthritis . The past decade has been an exciting period for clinical research and patient care in rheumatoid arthritis . This is mostly due to targeted biologic agents that have changed the outcome of this disease . DB08904 ( Cimzia(®) , UCB Inc. , GA , USA ) , which targets P01375 -α with a different mechanism of action than widely used biologics , was initially investigated for Crohn 's disease but has now been shown to be effective for rheumatoid arthritis . There have been three significant clinical trials demonstrating the efficacy of certolizumab pegol in active rheumatoid arthritis ; two with combination methotrexate and one with monotherapy . This article will summarize the data from those trials and compare some of the characteristics of certolizumab pegol to conventional disease-modifying antirheumatic drugs and other biologic agents . Treatment recommendations are beyond the scope of this review ; however , with many options available , there will be annotations on current trends in the care of this chronic disease . DB08904 : an evidence-based review of its place in the treatment of Crohn 's disease . INTRODUCTION : Crohn 's disease ( CD ) is a chronic inflammatory bowel disease characterized by a relapsing/remitting course with transmural inflammation of potentially any section of the digestive tract . DB08904 ( CZP ) is a pegylated Fc-free Fab ' fragment of a humanized anti- P01375 -alfa monoclonal antibody that is in development for clinical use in CD . AIMS : To review the available data with CZP in CD , to investigate its possible place in therapy . EVIDENCE REVIEW : Available studies suggest that CZP has the potential to achieve and maintain clinical response and remission in moderate to severe CD , and to improve quality of life compared with placebo . Further studies with CZP are also ongoing . PLACE IN THERAPY : Although only suggested by currently available studies , successive clinical practice and further ongoing trials may confirm a positive role for CZP as a new anti- P01375 treatment in CD . The impact on clinical management or on resources can not be estimated until the results from all phase III clinical trials are available and the price is determined . Efficacy and safety of certolizumab pegol in rheumatoid arthritis : meeting rheumatologists ' requirements in routine clinical practice . DB08904 , a pegylated Fab ' anti-tumour necrosis factor ( P01375 ) -α agent , has shown efficacy in patients with rheumatoid arthritis ( RA ) unresponsive to previous treatment . In key randomised controlled trials involving patients with moderate to severe RA and an inadequate response to methotrexate or one or more disease-modifying antirheumatic drug ( DMARD ) , the efficacy of certolizumab pegol , as monotherapy or with methotrexate , was similar to that reported in other anti- P01375 clinical studies , with 60 % or fewer of patients achieving American College of Rheumatology 20 % improvement in RA . Rapid clinical response was also seen , with significant differences evident at week 1 , and efficacy maintained at study end and in open-label extensions . Adding certolizumab pegol to non-biological DMARDs is efficacious in other RA populations . In the CERTAIN study , certolizumab increased remission rates , prevented disease worsening and improved work productivity and daily activity in patients with low to moderate RA . In the REALISTIC study , rapid and consistent clinical responses were observed in a diverse group of anti- P01375 -eligible RA patients representing those seen in clinical practice . In the RAPID studies , rapid and sustained reduction in RA signs and symptoms , inhibition of structural joint damage progression , and improved physical function were seen with certolizumab pegol plus methotrexate versus methotrexate alone in RA patients with an incomplete response to methotrexate . DB08904 was generally well-tolerated in clinical trials , although long-term observational data are not yet available . Current data suggest that certolizumab pegol suits a ' treat to target ' approach , providing rapid and sustained improvements in RA signs and symptoms , and beneficial effects on workplace and home productivity in patients with RA . Resistance to killing by tumor necrosis factor in an adipocyte cell line caused by a defect in arachidonic acid biosynthesis . We have found that Q96RJ0 -R6 , which are resistant to the cytotoxic effects of tumor necrosis factor ( P01375 ) in the presence of cycloheximide ( Reid , T. R. , Torti , F. , and Ringold , G. M. ( 1989 ) J. Biol. Chem. 264 , 4583-4589 ) , have reduced ability to release arachidonic acid ( 20:4 ) from membrane phospholipids in response to either P01375 or the calcium ionophore A23187 treatment . However , no defect in the activity of phospholipase A2 , the principal enzyme responsible for the release of 20:4 from phospholipids , was observed in these cells . Detailed biochemical characterization of these P01375 -resistant cells has revealed that these cells are unable to synthesize 20:4 endogenously because of a defect in delta 6-desaturase , the rate-limiting enzyme of 20:4 biosynthesis . This deficiency leads to a marked decrease in the steady-state levels of 20:4 present in choline-containing phospholipid ( PC ) and ethanolamine-containing phospholipid ( PE ) . The Q96RJ0 -R6 cells , however , are capable of incorporating exogenous 20:4 into PC and PE , and when loaded in such manner they become significantly more sensitive to the cytotoxic effects of P01375 in the presence of cycloheximide . Therefore , the release of arachidonic acid from phospholipids appears to be a critical element in the signaling pathway utilized by P01375 and is essential to the rapid cytotoxic response elicited by P01375 in the absence of protein synthesis in wild-type Q96RJ0 cells . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . DB08904 attenuates the pro-inflammatory state in endothelial cells in a manner that is atheroprotective . OBJECTIVES : Rheumatoid arthritis ( RA ) is associated with accelerated atherosclerosis and premature cardiovascular death . Anti- P01375 therapy is thought to reduce clinical cardiovascular disease risk and improve vascular function in RA patients . However , the specific effects of P01375 inhibitors on endothelial cell function are largely unknown . Our aim was to explore the effects of certolizumab pegol ( CZP ) on P01375 -activated human aortic endothelial cells ( HAoECs ) . METHODS : HAoECs were cultured in vitro and exposed to i ) P01375 alone , ii ) P01375 plus CZP , or iii ) neither agent . Microarray analysis and quantitative polymerase chain reaction were used to analyse gene expression . Activation of NF-κB was investigated using immunocytochemistry , high content analysis and western blotting . Flow cytometry was performed to detect microparticle release from HAoECs . RESULTS : P01375 alone had strong effects on endothelial gene expression , while P01375 and CZP together produced a global gene expression pattern similar to untreated controls . In particular , genes for P16581 , P19320 and P05362 were significantly up-regulated by P01375 treatment . Notably , the P01375 /CZP cocktail prevented the up-regulation of these genes . P01375 -induced nuclear translocation of NF-κB was abolished by treatment with CZP . In addition the increased production of endothelial microparticles in P01375 -activated HAoECs was prevented by treatment with CZP . CONCLUSIONS : We have found at cellular level , that a clinically available P01375 inhibitor , CZP i ) reduces adhesion molecule expression ; ii ) prevents P01375 -induced activation of the NF-κB pathway and iii ) prevents the production of microparticles by activated endothelial cells . This could be central to the prevention of inflammatory environments underlying these conditions . Current directions of biologic therapies in inflammatory bowel disease . Crohn 's disease ( CD ) and ulcerative colitis ( UC ) are chronic inflammatory bowel diseases which can be difficult to control with conventional therapies . A greater understanding of their pathophysiology has led to new therapies that target specific molecules of the inflammatory cascade . Three anti-tumor necrosis factor ( P01375 ) monoclonal antibodies have been developed . DB00065 and adalimumab can induce clinical response and sustained remission in CD . DB00065 is also effective in UC . DB08904 gives good short-term results but long-term efficacy has yet to be determined in other clinical trials . Therapies that target leucocyte trafficking ( anti-integrins ) have also been developed and are associated with good clinical response in CD . DB00108 ( anti-α4 integrin antibody ) is associated with important side effects and is not used anymore in gastroenterology in Europe but is still used in the USA . DB09033 ( MLN0002 ) , an anti-α4β7 integrin antibody , has a good efficacy and safety profile . Monoclonal antibodies targeting other cytokines are also under development . For example , ustekinumab ( CNTO 1275 ) inhibits interleukins 12 and 23 . It is associated with a good clinical response in CD . New concepts in anti-tumor necrosis factor therapy for inflammatory bowel disease . Crohn 's disease is a T helper type 1 response immune disease characterized by increased production of interleukin-12 tumor necrosis factor-a ( P01375 ) , and interferon-g . Clinical trials have demonstrated that inhibition of P01375 is effective for the treatment of Crohn 's disease . Adverse events reported in patients treated with anti- P01375 agents include immunogenicity , acute infusion reactions , delayed hypersensitivity-type reactions , autoimmune diseases including drug-induced lupus and demyelination , and infection . This article reviews new concepts in the treatment of Crohn 's disease and ulcerative colitis with a variety of anti- P01375 biologic therapies : infliximab , adalimumab , DB08904 , CDP571 , etanercept , and onercept . The efficacy and safety of certolizumab pegol ( CZP ) in the treatment of active rheumatoid arthritis ( RA ) : a meta-analysis from nine randomized controlled trials . OBJECTIVE : DB08904 ( CZP ) is a novel anti- P01375 agent that is used for patients with moderate to severe active rheumatoid arthritis ( RA ) . However , the efficacy of CZP in RA remains controversial . Thus , we performed this meta-analysis to assess the efficacy and safety of CZP in the treatment of RA patients . METHODS : Eligible studies were randomized controlled trials ( RCTs ) that evaluated the efficacy and safe of CZP in the patients with active RA . The primary outcome was American College of Rheumatology 20 % ( ACR20 ) , and secondary outcome were ACR50 , ACR70 , disease activity , patient-reported outcomes ( PROs ) , and adverse events . A fixed-effect model or random-effect model was used to pool the estimates , depending on the absence or presence of heterogeneity among the included studies . RESULTS : Nine RCTs with a total of 5228 patients were included in this meta-analysis , and all of the patients were administered CZP or placebo . The pooled results showed that CZP significantly improved the ACR20 , ACR50 , ACR70 response rates , and physical function . CZP was associated with a statistically significant reduction in Disease Activity Score in 28 joints-Erythrocyte sedimentation rate , arthritis pain , and fatigue . Patients who received CZP treatment did not have a higher incidence of treatment-related adverse events , no matter in any intensity . CONCLUSIONS : CZP 200 or 400mg in the treatment of active RA significantly reduced the RA signs and symptoms , and improved physical function as compared with the placebo . More large-scale RCTs are needed to evaluate the long-term efficacy and safety of CZP in the treatment of active RA . [ Treatment of patients with destructive arthritis with certolizumab pegol ] . DB08904 is a new anti- P01375 inhibitor which has been approved for the treatment of rheumatoid arthritis since October 2009 . Due to the modification of the antibody fragment by the adherence of polyethylene glycol ( PEG ) a sufficient distribution in inflammatory tissue was found in animal experiments . In two individual case reports a remission of therapy refractive arthritis was achieved by administration of certolizumab pegol . Optimizing anti-tumor necrosis factor strategies in inflammatory bowel disease . The introduction of infliximab , a mouse/human chimeric monoclonal antibody to tumor necrosis factor ( P01375 ) , is an important advance in the treatment of Crohn 's disease . DB00065 is effective for induction and maintenance of remission in patients with inflammatory luminal and fistulizing disease . The development of human antichimeric antibodies ( HACAs ) has led to infusion reactions and loss of efficacy in patients treated with infliximab . Strategies to reduce the frequency of HACA formation include induction of immunologic tolerance with a three-dose regimen at 0 , 2 , and 6 weeks followed by systematic maintenance dosing every 8 weeks ; concomitant immunosuppressive therapy with azathioprine , 6-mercaptopurine , or methotrexate ; and premedication with intravenous corticosteroids . Humanized or fully human anti- P01375 biotechnologic agents , including CDP571 , DB08904 , etanercept , adalimumab , and onercept , are theoretically less immunogenic than the chimeric antibody infliximab . DB00005 is not effective for Crohn 's disease . CDP571 is not effective in unselected patients with active Crohn 's disease , but it may be effective in patients with elevated P02741 . The efficacy of DB08904 , adalimumab , and onercept is under investigation . The different mechanisms of action of these anti- P01375 agents may account for their variable efficacy . Their benefits , however , must be considered in the context of their risks , including infusion reaction ; delayed hypersensitivity-like reaction ; new onset of autoimmunity , with rare cases of drug-induced lupus and new-onset demyelination ; and the potential for rare but serious infections . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Treatment of Crohn 's disease with certolizumab pegol . Biologic therapies have revolutionized the treatment of Crohn 's disease ( CD ) . Targeting P01375 with monoclonal antibodies has changed the therapeutic landscape for tackling refractory and complicated CD . Intravenous use of infliximab , a chimeric monoclonal antibody to P01375 is , however , limited by the occurrence of adverse events , infusion reactions , infectious complications , aggravation of heart failure , the occurrence of neurological demyelinating conditions and induction of rare malignancies . The incremental development of next-generation P01375 antibodies and binding proteins through antibody-engineering techniques has followed , with the aim of producing efficacious drugs that are less expensive to produce , have a convenient route of administration and have fewer side effects . DB08904 ( DB08904 , Cimzia ) is an engineered humanized anti- P01375 antibody Fab fragment that minimizes the protein component and is conjugated to polyethylene glycol . Clinical studies have demonstrated efficacy in the treatment of moderate-to-severe active CD . Reported adverse events in the clinical trial program have been largely of mild-to-moderate severity , and occurred at similar frequencies in the active-treatment and placebo groups . DB08904 will be a useful addition to the armamentarium of biologic agents that can be used for the long-term treatment of CD . Inorganic lead enhances cytokine-induced elevation of matrix metalloproteinase P14780 expression in glial cells . Inorganic lead ( Pb ) is a ubiquitous environmental contaminant that produces a variety of deleterious effects in the central nervous system ( CNS ) . Matrix metalloproteinases ( MMPs ) , specifically P14780 , induced by inflammatory cytokines , are increasingly being implicated in CNS pathology . The present study demonstrates that low concentrations of either pro-inflammatory cytokines ( P01375 and IL-1beta ) or Pb did not influence the P14780 expression in a glial cell line ( P13671 ) when added separately . However , combined administration of Pb and cytokines induced a marked synergized elevation of P14780 expression in spite of a reduction in the number of glial cells . These results demonstrate a possible new mechanism by which Pb may induce neuropathological processes . The clinical efficacy and safety of certolizumab pegol in rheumatoid arthritis . IMPORTANCE OF THE FIELD : The treatment of rheumatoid arthritis has changed dramatically over the past 25 years , first with the introduction of methotrexate and then the introduction of biologic therapy . These agents have provided patients with multiple treatment options to try to achieve disease remission . Unfortunately , no one single agent is fully effective in every patient ; different patients respond to different therapies , even those with the same mechanism of action , in different ways . Another medication , such as certolizumab pegol , is a welcome addition to our treatment armamentarium of rheumatoid arthritis . AREAS COVERED IN THIS REVIEW : The basis of this review is all the peer-reviewed manuscripts found in PubMed and Medline searches from 1990 to 2009 and abstracts on certolizumab pegol presented at the American College of Rheumatology and European League Against Rheumatism within the past 5 years . WHAT THE READER WILL GAIN : This review should enable the reader to fully understand the benefit:risk ratio of certolizumab pegol in the treatment of rheumatoid arthritis . TAKE HOME MESSAGE : DB08904 is an effective agent either in combination with methotrexate or as monotherapy in the treatment of rheumatoid arthritis with a safety profile similar to other approved P01375 inhibitors . [ DB08904 ] . DB08904 is a new anti- P01375 drug formed by the Fab ' fragment of a humanized mouse monoclonal antibody bound to two molecules of polyethylene glycol . DB08904 recognizes and binds to human P01375 -α , both in its soluble and membrane bound form , and has shown clinical efficacy in controlled trials for the treatment of RA and Crohns ' disease . In this review we summarize the structural characteristics and clinical efficacy data , as well as safety data of this anti- P01375 agent in patients with RA . DB08904 for the treatment of Crohn 's disease . In this article we provide a contemporary overview of available clinical data on certolizumab pegol , a pegylated anti-tumor necrosis factor ( P01375 ) alpha agent that comprises a uniquely small protein , and its emerging role as a therapy for Crohn 's disease ( CD ) . The results from a comprehensive clinical trial program suggest that certolizumab pegol offers rapid and sustained remission of moderate to severe CD . DB08904 is an effective and well-tolerated therapy both in patients who have already received biologics and in patients who are anti- P01375 naïve . Benefits of therapy include a stable dosing regimen , which allows for rapid induction of a clinical response followed by long-term maintenance of response and remission under one fixed dose . Treatment with certolizumab pegol has been shown to improve function and quality of life in patients with CD , and insights into the potential mechanisms by which certolizumab pegol effects a response in CD suggest that this agent may have the potential to slow or even modify disease progression . Early therapy is particularly effective and could help control CD progression and lessen the burden of disease on patients . The cytotoxic effects of certolizumab pegol and DB06674 mediated by transmembrane tumor necrosis factor α . BACKGROUND : Anti-tumor necrosis factor α ( anti- P01375 -α ) agents have been successfully applied for the treatment of rheumatoid arthritis , Crohn 's disease , and other chronic inflammatory diseases . Not only the neutralization of soluble P01375 -α but also the effect on transmembrane P01375 -α is important mechanisms of action of anti- P01375 -α agents . This study investigated the cytotoxic effects of new anti- P01375 -α agents , certolizumab pegol and DB06674 , which are mediated by transmembrane P01375 -α . METHODS : Transmembrane P01375 -α-expressing Jurkat T cells that did not express P01375 receptors were used . The binding ability of each anti- P01375 -α agent to transmembrane P01375 -α , antibody-dependent cell-mediated cytotoxicity , complement-dependent cytotoxicity , and the apoptotic effect were examined . RESULTS : DB08904 and DB06674 bound to transmembrane P01375 -α . Golimumab induced antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity , which was comparable to infliximab and adalimumab . However , certolizumab pegol did not induce antibody-dependent cell-mediated cytotoxicity or complement-dependent cytotoxicity . DB08904 directly induced nonapoptotic cell death in transmembrane P01375 -α-expressing cells . Golimumab induced a weaker apoptotic effect than infliximab and adalimumab . CONCLUSIONS : The cytotoxic effects of anti- P01375 -α agents on P01375 -α-expressing cells are considered to be associated with the clinical effect of these agents on granulomatous diseases . The direct cytotoxic effect of certolizumab pegol on P01375 -α-producing cells may contribute to its clinical efficacy in Crohn 's disease . Golimumab may be less effective for granulomatous diseases . Strategies for targeting tumour necrosis factor in Q9UKU7 . Tumour necrosis factor ( P01375 ) plays an important role in mediating the inflammation of inflammatory bowel disease , in particular , Crohn 's disease . Strategies aimed at reducing tumour necrosis factor in patients with inflammatory bowel disease include the mouse/human chimeric monoclonal antibody infliximab , the humanized monoclonal antibody CDP571 , the human soluble P01375 p55 receptor onercept , the human monoclonal antibody D2E7 ( adalimumab ) , the anti- P01375 human antibody Fab ' fragment-polyethelene glycol ( PEG ) conjugate DB08904 , and the small molecules thalidomide and CNI-1493 ( Q96HU1 -kinase inhibitor ) . DB00065 is effective for treating active Crohn 's disease , maintaining remission , closing fistulas , maintaining fistula closure , and treating ankylosing spondylitis . DB00065 is also being investigated for the treatment of ulcerative colitis . Side-effects occurring in patients treated with infliximab include human anti-chimeric antibodies , infusion reactions , delayed hypersensitivity reactions , formation of autoantibodies , and , in rare circumstances , drug-induced lupus and serious infections , including tuberculosis . CDP571 is effective for treating active Crohn 's disease , steroid sparing , and possibly for closing fistulas and maintaining remission . Side-effects occurring in patients treated with CDP571 include anti-idiotype antibodies , infusion reactions and the formation of autoantibodies . A controlled trial of etanercept in patients with Crohn 's disease was negative . Pilot studies with onercept , thalidomide , and CNI-1493 have suggested benefit for Crohn 's disease . There are no published data on the efficacy of adalimumab ( D2E7 ) or DB08904 for either Crohn 's disease or ulcerative colitis . Anti-tumour necrosis factor therapies are effective for the treatment of Crohn 's disease and are being investigated for ulcerative colitis . The PRECiSE 2 trial of certolizumab pegol , a new PEGylated anti- P01375 agent , in the treatment of Crohn 's disease : An interview with David A Schwartz , 13 June 2007 . DB08904 ( CDP 870 ) is a new anti-tumor necrosis factor ( P01375 ) therapy currently in development for the treatment of Crohn 's disease , rheumatoid arthritis , and psoriasis . DB08904 is the first PEGylated biologic anti- P01375 agent and has a high binding affinity for P01375 . Dr. Schwartz was an investigator of the PRECiSE ( PEGylated Antibody Fragment Evaluation in Crohn 's Disease Safety and Efficacy ) 2 trial of certolizumab pegol in patients with Crohn 's disease . DB08904 in Crohn 's disease . DB08904 is a humanized Fab ' fragment monoclonal antibody to tumor necrosis factor alpha ( P01375 ) . PEGylation increases its half-life , and it is administered subcutaneously to treat immune-mediated inflammatory diseases such as Crohn 's disease and rheumatoid arthritis . DB08904 improves quality of life and reduces clinical disease activity . Inflammatory markers such as P02741 ( CRP ) also decrease after administration of certolizumab pegol . The dose for induction of remission is 400 mg subcutaneously at weeks 0 , 2 and 4 . The dose for maintenance of remission is 400 mg sc given every four weeks . The safety profile is comparable with other anti- P01375 agents , and the major adverse events are related to infections . This article reviews the published data regarding the efficacy and safety of certolizumab pegol . Infectious complications of tumor necrosis factor blockade . PURPOSE OF REVIEW : Our understanding of the infection risks posed by tumor necrosis factor ( P01375 ) antagonists has continued to evolve in the 10 years since these drugs were first introduced . This review summarizes recent data regarding infection risk , examines potential structure-function relationships that may account for the differences , and discusses their implications with regard to tuberculosis prevention and management . RECENT FINDINGS : Recent prospective studies have confirmed the risk of tuberculosis reactivation posed by P01375 antibodies to be several fold greater than soluble P01375 receptor . DB08904 , a monovalent anti- P01375 Fab ' fragment appears to share this risk , despite its lack of Fc and its inability to cross-link transmembrane P01375 . Screening and initiation of treatment for latent tuberculosis ( TB ) infection can greatly reduce the TB risk of anti- P01375 treatment in western countries . However , alternative strategies to prevent TB because of new transmission may be required as these therapies become available worldwide . Current recommendations for withdrawal of anti- P01375 therapy when TB is diagnosed place patients at risk for paradoxical worsening because of recovery of P01375 -dependent inflammation . Recent case reports suggest reinstitution of P01375 blockade may be safe and effective adjunctive treatment in such cases , but prospective studies are needed to confirm these observations . SUMMARY : P01375 blockers have transformed treatment of several chronic inflammatory conditions . Further research is needed to determine how best to prevent and manage their infectious complications and to determine their potential adjunctive therapeutic role in chronic infection diseases . Meta-analysis : the efficacy and safety of certolizumab pegol in Crohn 's disease . BACKGROUND : DB08904 is the third anti- P01375 agent approved by the Food and Drug Administration of the United States . AIM : To provide a comprehensive up-to-date review of the efficacy and safety of certolizumab in Crohn 's disease ( CD ) . METHODS : Electronic databases , including PubMed , EMBASE , the Cochrane library and the Science Citation Index , were searched to retrieve relevant trials . In addition , meeting abstracts and the reference lists of retrieved articles were reviewed for further relevant studies . RESULTS : Three trials , enrolling a total of 1040 patients , are included in the meta-analysis to evaluate the short-term efficacy of certolizumab , which is effective for rapid induction and long-term maintenance of clinical response or remission and can improve quality of life in patients with Crohn 's disease . Certolizumab is also effective for patients who have lost response to infliximab . However , its efficacy in infliximab-exposed patients is probably less than in infliximab-naive patients . Re-induction with certolizumab in patients who have flared on maintenance therapy can rescue a significant proportion of patients . There is no significant association between the efficacy of certolizumab and the baseline P02741 level . In comparison with placebo , certolizumab does not increase the risk of serious adverse events . CONCLUSIONS : Certolizumab is effective and safe in treating Crohn 's disease . Further studies are still required to assess its full safety profile . Evaluation of pharmacokinetics and pharmacodynamics and clinical efficacy of certolizumab pegol for Crohn 's disease . INTRODUCTION : P01375 -α antagonists have transformed the treatment of patients with Crohn 's disease ( CD ) . DB08904 ( CZP ) is the third P01375 -α antagonist to be approved for use in the United States but is not currently approved in Europe . AREAS COVERED : This review evaluates the pharmacokinetics , pharmacodynamics and efficacy of CZP in CD . Safety , immunogenicity and its use in pregnancy have also been assessed . A literature search was conducted using Pub Med ( 2004 - 2014 ) for the terms ' Crohn 's disease ' and ' certolizumab pegol ' or ' certolizumab ' or ' cimzia ' . Additional studies were identified from other sources including citation . EXPERT OPINION : As a Fab ' fragment , CZP is effective in binding P01375 -α , but does not cause Fc-mediated effects . PEGylation has improved its pharmacokinetic profile and allowed for an increased half-life of 2 weeks . Benefit for inducing response ( an improvement in symptoms ) and maintenance of remission has been shown . However , the benefit is less clear for the more stringent end-points of inducing remission and mucosal healing . There may be an advantage from the PEGylated formulation of CZP in terms of reduced injection-site reactions , reduced placental transfer in pregnancy and as a treatment option in patients who are unable to tolerate infliximab . DB08904 and rheumatoid arthritis . Just another P01375 alpha antagonist , no therapeutic advantage . No comparison with other P01375 alpha antagonists ; possible bleeding risk . The use of P01375 blocking agents in rheumatoid arthritis : an overview . P01375 has been found to play a pivotal role in the pathogenic mechanisms of rheumatoid arthritis ( RA ) . The overexpression of P01375 in RA synovium , the data from in vitro synovial cell cultures with the use of anti- P01375 antibody and the results from P01375 blockade in animal models of arthritis argued for the importance of this cytokine in RA . Drugs targeting P01375 have been developed to neutralise the deleterious effects of this inflammatory cytokine . There are currently three drugs available in the treatment of RA patients with active disease , which was refractory to conventional treatments including methotrexate , infliximab ( a chimeric mouse/human monoclonal antibody ) , etanercept ( a fusion protein combining 2 p75 P01375 receptors with a Fc fragment of human IgG ( 1 ) ) and adalimumab ( a fully human monoclonal antibody ) . These three drugs have proved to be effective and safe in appropriate and well conducted clinical trials and showed effectiveness in slowing and even arresting the progression of radiographic damage . With the long-term surveillance of these drugs serious adverse events were described , particularly intracellular organism infections such as tuberculosis . Other drugs targeting P01375 are in development and include monoclonal antibody ( CDP571 ) , pegylated molecules ( DB08904 and PEG-r-Hu-sTNF-RI ) or soluble p55 P01375 receptor construct ( lenercept ) . These new biological therapies blocking P01375 undoubtedly constitute a considerable advance in the management of RA , but careful evaluation at the initiation of the treatment and long-term surveillance of the patients receiving such drugs remains necessary . Pegylation of biological molecules and potential benefits : pharmacological properties of certolizumab pegol . PEGylation of biological proteins , defined as the covalent conjugation of proteins with polyethylene glycol ( PEG ) , leads to a number of biopharmaceutical improvements , including increased half-life , increased solubility and reduced aggregation , and reduced immunogenicity . Since their introduction in 1990 , PEGylated proteins have significantly improved the management of various chronic diseases , including rheumatoid arthritis ( RA ) and Crohn 's disease . DB08904 is the only PEGylated anti-tumour necrosis factor ( P01375 ) -α agent . It is a PEGylated , humanised , antigen-binding fragment of an anti- P01375 monoclonal antibody . Unlike other anti- P01375 agents , it has no crystallisable fragment ( Fc ) domain . Because of its novel structure , certolizumab pegol may have a different mechanism of action to the other anti- P01375 agents , and also has different pharmacodynamic properties , which could possibly translate to a different safety profile . Pharmacodynamic studies have shown that certolizumab pegol binds to P01375 with a higher affinity than adalimumab and infliximab . DB08904 is also more potent at neutralising soluble P01375 -mediated signalling than adalimumab and infliximab , and has similar or lesser potency to etanercept . DB08904 does not cause detrimental in vitro effects such as degranulation , loss of cell integrity , apoptosis , complement-dependent cytotoxicity and antibody-dependent cell-mediated cytotoxicity . DB08904 may also penetrate more effectively into inflamed arthritic tissue than other anti- P01375 agents , and is not actively transported across the placenta during pregnancy . Pharmacokinetic studies in healthy volunteers demonstrated that single intravenous and subcutaneous doses of certolizumab pegol had predictable pharmacokinetics . The pharmacokinetics of certolizumab pegol in patients with RA and Crohn 's disease were consistent with pharmacokinetics in healthy volunteers . DB08904 -- what role does this new P01375 inhibitor have in the treatment of RA ? The efficacy and safety of a new tumor necrosis factor inhibitor , certolizumab pegol , in active rheumatoid arthritis has now been assessed in three phase III , multicenter , randomized , double-blind , placebo-controlled clinical trials . This commentary focuses on the paper by Keystone et al. , in which patients were followed for the longest duration . This study , which compared two doses of subcutaneous certolizumab pegol with placebo in patients with active RA receiving methotrexate , showed no advantage of 400 mg over 200 mg certolizumab pegol over 52 weeks , after induction with 400 mg . The nature of the patients enrolled in this study , trial design and possible safety issues are discussed , as is whether this trial can teach us anything about tumor necrosis factor inhibitors in general . On the basis of the results from this study , certolizumab pegol does not represent a major addition to our armamentarium , but because of the slightly different mechanism of action and structure of this drug , and the apparently acceptable therapeutic effects over one year , it is , nevertheless , welcome . Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing P04626 . In an attempt to treat cancer patients with P04626 overexpressing tumors , we developed a chimeric antigen receptor ( CAR ) based on the widely used humanized monoclonal antibody ( mAb ) DB00072 ( Herceptin ) . An optimized CAR vector containing P10747 , 4-1BB , and CD3zeta signaling moieties was assembled in a gamma-retroviral vector and used to transduce autologous peripheral blood lymphocytes ( PBLs ) from a patient with colon cancer metastatic to the lungs and liver , refractory to multiple standard treatments . The gene transfer efficiency into autologous T cells was 79 % CAR(+) in CD3(+) cells and these cells demonstrated high-specific reactivity in in vitro coculture assays . Following completion of nonmyeloablative conditioning , the patient received 10(10) cells intravenously . Within 15 minutes after cell infusion the patient experienced respiratory distress , and displayed a dramatic pulmonary infiltrate on chest X-ray . She was intubated and despite intensive medical intervention the patient died 5 days after treatment . Serum samples after cell infusion showed marked increases in interferon-gamma ( P01579 ) , granulocyte macrophage-colony stimulating factor ( GM- P04141 ) , tumor necrosis factor-alpha ( P01375 ) , interleukin-6 ( P05231 ) , and P22301 , consistent with a cytokine storm . We speculate that the large number of administered cells localized to the lung immediately following infusion and were triggered to release cytokine by the recognition of low levels of P04626 on lung epithelial cells . DB00173 nucleotides inhibit cytokine generation by human mast cells through a Gs-coupled receptor . DB00171 and ADP activate functionally distinct G protein-coupled purinergic ( P2Y ) receptors . We determined the expression and function of adenine nucleotide-specific P2Y receptors on cord blood-derived human mast cells ( hMCs ) . Human MCs expressed mRNA encoding the ADP-specific P47900 , Q9H244 , and Q9BPV8 receptors ; the DB00171 /UTP-specific P41231 receptor ; and the DB00171 -selective Q96G91 receptor . ADP ( 0.05-50 muM ) induced calcium flux that was completely blocked by a P47900 receptor-selective antagonist and was not cross-desensitized by DB00171 . Low doses of ADP induced strong phosphorylation of P29323 and p38 MAPKs ; higher doses stimulated eicosanoid production and exocytosis . Although MAPK phosphorylation was blocked by a combination of P47900 - and Q9H244 -selective antagonists , neither interfered with secretion responses . Unexpectedly , both ADP and DB00171 inhibited the generation of P01375 in response to the O60603 ligand , peptidoglycan , and blocked the production of P01375 , P10145 , and MIP-1beta in response to leukotriene D(4) . These effects were mimicked by two DB00171 analogues , adenosine 5'-O-(3-thiotriphosphate) and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate ( BzATP ) , but not by adenosine . ADP , DB00171 , adenosine 5'-O-(3-thiotriphosphate) , and 2',3'-O-(4-benzoyl-benzoyl) adenosine 5'-triphosphate each induced DB02527 accumulation , stimulated the phosphorylation of CREB , and up-regulated the expression of inducible DB02527 early repressor , a CREB-dependent inhibitor of cytokine transcription . Human MCs thus express several ADP-selective P2Y receptors and at least one G(s)-coupled ADP/ DB00171 receptor . Nucleotides could therefore contribute to MC-dependent microvascular leakage in atherosclerosis , tissue injury , and innate immunity while simultaneously limiting the extent of subsequent inflammation by attenuating the generation of inducible cytokines by MCs . DB08904 compared to natalizumab in patients with moderate to severe Crohn 's disease : results of a decision analysis . INTRODUCTION : A significant proportion of patients with Crohn 's disease ( CD ) lose response to antibodies directed against tumor necrosis factor α ( P01375 ) . Prior P01375 -antagonist failure is associated with lower rates of response to subsequent P01375 -antagonist therapy . In patients failing two anti- P01375 agents , a choice exists between using a third-anti- P01375 therapy or natalizumab ( NAT ) , an α-4 integrin inhibitor . A cost-effectiveness analysis comparing these competing strategies has not been performed . METHODS : A decision analytic model was constructed to compare the performance of certolizumab pegol ( CZP ) versus NAT in patients with moderate to severe CD . Previously published estimates of efficacy of third-line anti- P01375 therapy and NAT were used to inform the model . Costs were expressed in 2010 US dollars . A 1-year time frame was used for the analysis . RESULTS : In the base case estimate , use of NAT was only marginally more effective [ 0.71 vs. 0.70 quality adjusted life-years ( QALYs ) ] than CZP but was expensive with an incremental cost-effectiveness ratio ( Q03060 ) of $ 381,678 per QALY gained . For CZP 2 months response rate of at least 24 % , NAT had an Q03060 above the willingness-to-pay ( WTP ) threshold . The model was sensitive to the costs of both therapies ; for all CZP costs below $ 2,300 per dose , NAT had higher Q03060 than the WTP threshold . Substituting adalimumab for CZP resulted in similar Q03060 estimates and thresholds for NAT use . CONCLUSIONS : In patients with moderate to severe CD failing two P01375 -antagonists , using a third P01375 -antagonist therapy appears to be a cost-effective strategy without significantly compromising treatment efficacy . Prasugrel : a new antiplatelet drug for the prevention and treatment of cardiovascular disease . Prasugrel , trade name DB06209 , is an investigational new antiplatelet drug currently under review for clinical use by the Food and Drug Administration . It is a thienopyridine analog with a structure similar to that of clopidogrel and ticlopidine . Thienopyridine derivatives inhibit platelet aggregation induced by adenosine diphosphate by irreversibly inhibiting the binding of adenosine diphosphate to the purinergic Q9H244 receptor on the platelet surface . Prasugrel has been shown to be a potent antiplatelet agent with a faster , more consistent , and greater inhibition of platelet aggregation compared with clopidogrel . It is debatable , however , how effectively these pharmacologic benefits will translate to clinical benefits . The results of the large TRITON-TIMI 38 trial , which compared prasugrel and clopidogrel in patients with acute coronary syndrome who were scheduled to receive coronary stents , demonstrated a significant reduction in ischemic events , including stent thrombosis , with prasugrel , but with an increased risk of major bleeding . The exact role of prasugrel in the management of ischemic heart disease is still being defined , but the risk:benefit ratio will likely play a major role in directing the best place for therapy with this new agent . DB08904 : a review of its use in the management of rheumatoid arthritis . DB08904 ( Cimzia(®) ) is a recombinant , polyethylene glycolylated , antigen-binding fragment of a humanized monoclonal antibody that selectively targets and neutralizes tumour necrosis factor ( P01375 ) -α . The drug is indicated for subcutaneous use every 2 or 4 weeks ( q2w or q4w ) for the treatment of adults with moderate to severe active rheumatoid arthritis ( RA ) . The efficacy of subcutaneous certolizumab pegol in adults with active RA has been investigated in several well designed , placebo-controlled trials . In four pivotal studies of ≤52 weeks duration , patients with moderate to severe disease receiving recommended dosages of certolizumab pegol ( 200 mg q2w or 400 mg q4w ) , either as monotherapy ( after failing prior disease-modifying anti-rheumatic drug [ DMARD ] therapy ) or in combination with methotrexate ( after responding inadequately to methotrexate alone ) , experienced rapid clinical improvement , with some combination trials also demonstrating inhibition of radiographic progression . The beneficial effects of certolizumab pegol therapy were generally maintained for up to ≈5 years in clinical trial extensions in which the drug was administered at dosages of 400 mg q4w or q2w . Additional studies suggest certolizumab pegol is also effective in patients who are Asian or have low to moderate disease activity , as well as more clinically representative patient populations . The tolerability profile of certolizumab pegol was acceptable , with infections/infestations the most common adverse events . Thus , certolizumab pegol is an effective option for the management of active RA in adults , although additional long-term and comparative efficacy and tolerability data are needed to help definitively position certolizumab pegol relative to other biological DMARDs , particularly other anti- P01375 agents . Profile of certolizumab and its potential in the treatment of psoriatic arthritis . Psoriatic arthritis ( PsA ) is a chronic inflammatory arthropathy associated with psoriasis ( PsO ) . PsA could be considered an enthesal disease because of the link between mechanical stress ( entheses ) and immunologically active tissue ( synovium ) . Evidence of efficacy of anti-tumor necrosis factor alpha ( P01375 -α ) is supported by reduction of histological vascularity and immune cell infiltrates in synovial tissue after treatment . DB08904 ( CZP ) is a polyethylene glycolylated ( PEGylated ) Fab ' fragment of a humanized monoclonal antibody that binds and neutralizes human P01375 -α . The PEG moiety of the Fab fragment , markedly increases the half-life of CZP and confers to the drug a unique structure that differs from the other anti- P01375 -α agents tested for the treatment of Crohn 's disease , rheumatoid arthritis , ankylosing spondylitis , axial spondyloarthritis , nonradiographic spondyloarthritis , PsO , and PsA . In contrast to other anti- P01375 -α agents , CZP did not mediate increased levels of apoptosis , suggesting that these mechanisms are not essential for the anti- P01375 -α efficacy in Crohn 's disease . As CZP , infliximab , and adalimumab , but not etanercept , almost completely inhibited lipopolysaccharide-induced interleukin-1 beta release from monocytes , this cytokine-production inhibition may be relevant for drug efficacy . Due to these characteristics , it has been demonstrated in clinical studies that CZP effectively improves signs and symptoms of arthritis and physical function and skin manifestations of PsO , with a safety profile similar to rheumatoid arthritis . This drug can be considered as a valid treatment in patients affected by PsA . The efficacy and tolerability profiles suggest CZP as a suitable antipsoriatic drug in the treatment of PsA . [ Biologic therapies in chronic inflammatory bowel diseases ] . Crohn 's disease and ulcerative colitis are chronic inflammatory bowel diseases which can be difficult to control with conventional therapies . Thanks to a better knowledge of their physiopathology , new therapies aimed at specific targets of the inflammatory cascade were developed . Three monoclonal anti- P01375 antibodies were produced . DB00065 and adalimumab , currently widely used , can induce sustained remission in Crohn 's disease . DB00065 is also efficacious in UC . DB08904 provides good short term results ; its long term efficacy , however , remains to be assessed by further clinical trials . Therapies targeting leucocyte trafficking ( anti-integrine ) have also been provided and are associated with good clinical responses in Crohn 's disease . DB00108 ( anti-alpha4 ) is responsible for significant side effects and is no longer in use in gasrtoenterology in Europe whereas MLN02 ( anti-alpha417 ) has a good profile in terms of efficacy and safety . Monoclonal anti bodies targeting other cytokines are under development , mainly ustekinumab which inhibits IL12 and IL23 . Ustekinumab generates favourable clinical responses in Crohn 's disease . The development of biologic therapies in inflammatory bowel disease has dramatically altered the course and management of these disorders . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . DB01393 restores the inhibition of DB00094 -induced follicular development and steroidogenesis by tumor necrosis factor-alpha through peroxisome proliferator-activated receptor-gamma pathway in an in vitro mouse preantral follicle culture . We recently reported that bezafibrate , a lipid-lowering drug of the fibrate class , administered in addition to clomiphene citrate ( CC ) successfully induced ovulation in CC-resistant polycystic ovary syndrome ( PCOS ) patients . We hypothesized that bezafibrate may directly affect ovarian follicle development . P01308 resistance and compensatory hyperinsulinemia are important for the pathogenesis of PCOS . In this study , we first examined the effects of tumor necrosis factor-alpha ( P01375 ) , which plays a role in insulin resistance , on follicle development by using the follicle culture system . P01375 significantly inhibited follicle-stimulating hormone ( DB00094 ) -induced follicle development , 17beta-estradiol ( E2 ) secretion , and ovulation rate in a dose-dependent manner . We then examined whether bezafibrate treatment could rescue the inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 . DB01393 treatment rescued inhibition of follicle development , secretion of E2 , and ovulation rate by P01375 . We examined the expression of peroxisome proliferator-activated receptor ( Q07869 ) subtypes in mouse preantral follicles . As the protein expression of only P37231 was observed in mouse preantral follicles , we examined whether bezafibrate could affect follicle development and steroidogenesis through P37231 pathways . Treatment with GW1929 , a selective P37231 agonist , restored inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 , whereas treatment with GW9662 , a selective P37231 antagonist , canceled the restorative effects of bezafibrate . Collectively , the results in this study suggest that bezafibrate may directly exhibit a restorative effect on the inhibition of ovarian follicle development and steroidogenesis by P01375 through the P37231 pathway .

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(Pro)renin promotes fibrosis gene expression in P29320 cells through a Nox4-dependent mechanism . The (pro)renin receptor ( PRR ) has recently been demonstrated to bind equally well renin and its precursor , prorenin , leading to a similar intracellular signaling independent of angiotensin II . In this study , we report that human embryonic kidney cells ( P29320 ) exposed to renin or prorenin for 24 h in the presence of a blocking concentration of the angtiotensin-converting enzyme inhibitor perindoprilate increased superoxide anion production as measured by luminescence ( lucigenin ) and electron spin resonance spectroscopy ( hydroxylamine radical transition ) . Also , both renin and prorenin increased Nox4 expression while Nox2 , p47(phox) , and p67(phox) remained unchanged . In an investigation of the effects of renin and prorenin on fibrosis genes , it appeared that both proteins stimulated transforming growth factor-β ( TGF-β ) , fibronectin , and plasminogen activator inhibitor type 1 ( P05121 ) expression and therefore participated to an overall switch toward a profibrotic state of the kidney cells . When the cells were transfected with a siRNA targeting the PRR , Nox4 expression was efficiently prevented as well as the increase in superoxide production , TGF-β , fibronectin , and P05121 . Finally , we demonstrated that transfection of the cells with a Nox4-specific small interfering ( si ) RNA also prevented fibrosis gene expression following treatment with renin or prorenin . The results demonstrate that renin and prorenin , through their specific membrane receptor and independently of angiotensin II , promote fibrosis gene expression via a Nox4-dependent mechanism . 20-Hydroxycholecalciferol , product of vitamin D3 hydroxylation by P450scc , decreases NF-kappaB activity by increasing IkappaB alpha levels in human keratinocytes . The side chain of vitamin D3 is hydroxylated in a sequential manner by cytochrome P450scc ( P05108 ) to form 20-hydroxycholecalciferol , which can induce growth arrest and differentiation of both primary and immortalized epidermal keratinocytes . Since nuclear factor-kappaB ( NF-kappaB ) plays a pivotal role in the regulation of cell proliferation , differentiation and apoptosis , we examined the capability of 20-hydroxycholecalciferol to modulate the activity of NF-kappaB , using DB00136 ( calcitriol ) as a positive control . 20-hydroxycholecalciferol inhibits the activation of NFkappaB DNA binding activity as well as NF-kappaB-driven reporter gene activity in keratinocytes . Also , 20-hydroxycholecalciferol induced significant increases in the mRNA and protein levels of the NF-kappaB inhibitor protein , IkappaB alpha , in a time dependent manner , while no changes in total NF-kappaB-p65 mRNA or protein levels were observed . Another measure of NF-kappaB activity , p65 translocation from the cytoplasm into the nucleus was also inhibited in extracts of 20-hydroxycholecalciferol treated keratinocytes . Increased IkappaB alpha was concomitantly observed in cytosolic extracts of 20-hydroxycholecalciferol treated keratinocytes , as determined by immunoblotting and immunofluorescent staining . In keratinocytes lacking vitamin D receptor ( P11473 ) , 20-hydroxycholecalciferol did not affect IkappaB alpha mRNA levels , indicating that it requires P11473 for its action on NF-kappaB activity . Comparison of the effects of calcitrol , hormonally active form of vitamin D3 , with 20-hydrocholecalciferol show that both agents have a similar potency in inhibiting NF-kappaB . Since NF-kappaB is a major transcription factor for the induction of inflammatory mediators , our findings indicate that 20-hydroxycholecalciferol may be an effective therapeutic agent for inflammatory and hyperproliferative skin diseases . Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation . Matrix metalloproteinases ( MMPs ) are essential for proper extracellular matrix remodeling , a process that takes place during obesity-mediated adipose tissue formation . Here , we examine expression profiles and the potential role of MMPs and their tissue inhibitors ( TIMPs ) in adipose tissue remodeling during obesity . Expression patterns are studied by Northern blot and real-time PCR in two genetic models of obesity ( ob/ob and db/db mice ) and in a diet-induced model of obesity ( AKR mice ) . Of the MMPs and TIMPs studied , mRNA levels for P08253 , P08254 , P39900 , P50281 , Q99542 , and P01033 are strongly induced in obese adipose tissues compared with lean tissues . In contrast , P09237 and P35625 mRNAs are markedly decreased in obesity . Interestingly , enzymatic activities of P39900 and of a new identified adipocyte-derived 30-kDa metalloproteinase are enhanced in obese adipose tissue fractions , demonstrating that MMP/ P01033 balance is shifted toward increased matrix degradation in obesity . Finally , we analyze the modulation of P08253 , Q99542 , and P01033 during 3T3- Q9NUQ9 preadipocyte differentiation , and we explore the effect of inhibition of MMP activity on in vitro adipogenesis . We find that the synthetic MMP inhibitor BB-94 ( DB03880 ) decreases adipose conversion of 3T3- Q9NUQ9 and primary rat preadipocytes . BB-94 represses differentiation without affecting mitotic clonal expansion but prevents the early expression of P17676 , a transcription factor that is thought to play a major role in the adipogenic program . Such findings support a role for the MMP/ P01033 system in the control of proteolytic events and adipogenesis during obesity-mediated fat mass development . Genetics of type 2 diabetes mellitus and other specific types of diabetes ; its role in treatment modalities . Type 2 diabetes mellitus ( T2DM ) is among the most challenging health issues of the 21st century and is associated with an alarming rise in the incidence . The pathophysiological processes that lead to development of T2DM are still unclear , however impairment in insulin secretion and/or action is clearly indicated . Type 2 diabetes is a polygenic disorder with multiple genes located on different chromosomes contributing to its susceptibility . Analysis of the genetic factors is further complicated by the fact that numerous environmental factors interact with genes to produce the disorder . Only a minority of cases of type 2 diabetes are caused by single gene defects and one example is maturity onset diabetes of the young ( MODY ) . Previous studies indicated that variants in genes encoding the pancreatic β-cell K+ DB00171 channel subunits Kir6.2 ( Q14654 ) and Q09428 ( Q09428 ) are associated with neonatal diabetes . Six different types of maturity onset diabetes of young ( MODY ) have been identified based on characteristic gene defect . The common Pro12Ala polymorphism in peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) gene was confirmed in several studies to be associated with type 2 diabetes as well . More recently , studies reported variants within a novel gene , Q9NQB0 , as a putative susceptibility gene for type 2 diabetes across many ethnic backgrounds around the world . MODY patients respond better to sulphonylureas and metformin , while neonatal diabetes patients with genetic mutations can be changed from insulin to oral drugs . We hereby provide a comprehensive review on the role of genetics in type 2 diabetes mellitus . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Extended kindred with recessive late-onset Alzheimer disease maps to locus 8p22- P38936 .2 : a genome-wide linkage analysis . Late-onset Alzheimer disease ( LOAD ) is a complex genetic disorder . Although genes involved in early-onset forms were discovered more than a decade ago , LOAD research has only been able to point out small effect loci , with the exception of P02649 . We mapped the gene predisposing to LOAD in an extended inbred family coming from a genetically isolated region ( 24 sampled individuals , 12 of whom are affected ) , completing a genome-wide screen with an Affymetrix10 K single nucleotide polymorphism microarray . Genotyping results were evaluated under model-dependent ( dominant and recessive ) and model-free analysis . We obtained a maximum nonparametric linkage score of 3.24 ( P=0.00006 ) on chromosome 8p22- P38936 .2 . The same genomic position also yielded the highest multipoint heterogeneity LOD ( HLOD ) under a recessive model ( HLOD=3.04 ) . When we compared the results of the model-dependent analysis , a higher score was obtained in the recessive model ( 3.04 ) than in the dominant model ( 1.0 ) . This is a new locus identified in LOAD , in chromosome 8p22- P38936 .2 and encompassing several candidate genes , among them P10909 and P48454 that were excluded by sequencing . The finding of a recessive model of inheritance , consistent with the assumption of inbreeding as a morbidity factor in this population , supports the notion of a role of recessive genes in LOAD . Caveolae and caveolin-1 are implicated in 1alpha,25(OH)2-vitamin D3-dependent modulation of Src , MAPK cascades and P11473 localization in skeletal muscle cells . We previously reported that DB00136 induces non-transcriptional rapid responses through activation of MAPKs in C2C12 skeletal muscle cells . However , there is little information on the molecular mechanism underlying the initiation of DB00136 signaling through this pathway . Plasma membrane components have been involved in some non-genomic effects . In this work , we investigated the role of caveolae and caveolin-1 ( cav-1 ) in DB00136 -stimulation of c-Src and MAPKs . When proliferating cells were pretreated with methyl beta cyclodextrin ( MbetaCD ) , a caveolae disrupting agent , under conditions in which cell morphology is not affected and no signs of apoptosis are observed , DB00136 -dependent activation of P27361 /2 , p38 MAPK and c-Src was suppressed . Similar results were obtained by siRNA technology whereby silencing of cav-1 expression abolished activation of c-Src and MAPKs induced by the hormone . By confocal immunocytochemistry it was observed that cav-1 colocalizes with c-Src in the periplasma membrane zone at basal conditions . Hormone treatment disrupted the colocalization of these proteins and redistributed them into cytoplasm and nucleus . Co-immunoprecipitation assays corroborated these observations . Changes in P11473 localization after DB00136 exposure were also investigated . Confocal microscopy images showed that the hormone induces P11473 translocation to the plasma membrane , and this effect is abolished by MbetaCD . Altogether , these data suggest that caveolae is involved upstream in c-Src-MAPKs activation by DB00136 and that P11473 and cav-1 participate in the rapid signaling elicited by the hormone . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . A tumor-infiltrating lymphocyte from a melanoma metastasis with decreased expression of melanoma differentiation antigens recognizes MAGE-12 . Twenty separate tumor infiltrating lymphocyte ( Q15399 ) bulk cultures and a tumor cell line were originated simultaneously from a fine needle aspiration biopsy of a metastasis in a patient with melanoma ( F001 ) previously immunized with the HLA-A*0201-associated gp100:209-217 ( 210 M ) peptide . None of the Q15399 recognized gp100 . However , 12 recognized autologous ( F001- P61006 ) and allogeneic melanoma cells expressing the HLA haplotype A*0201 , B*0702 , Cw*0702 . Further characterization of F001- P61006 demonstrated loss of gp100/PMel17 , severely decreased expression of other melanoma differentiation Ags and retained expression of tumor-specific Ags . Transfection of HLA class I alleles into B*0702/Cw*0702-negative melanoma cell lines identified HLA-Cw*0702 as the restriction element for F001- Q15399 . A cDNA library from F001- P61006 was used to transfect IFN-alpha-stimulated 293 human embryonal kidney ( 293- P29320 ) cells expressing HLA-Cw*0702 . A 100-gene pool was identified that induced recognition of 293- P29320 cells by F001- Q15399 . Subsequent cloning of the pool identified a cDNA sequence homologous , except for one amino acid ( aa 187 D --> A ) , to MAGE-12 . Among 25 peptide sequences from MAGE-12 with the HLA-Cw*0702 binding motif , MAGE-12:170-178 ( VRIGHLYIL ) induced P01579 release by F001- Q15399 when pulsed on F001-EBV-B cells at concentrations as low as 10 pg/ml . Peptide sequences from MAGE-1 , 2 , 3 , 4a , and 6 aligned to MAGE-12:170-178 were not recognized by F001- Q15399 . In summary a Q15399 recognizing a MAGE protein was developed from an HLA-A*0201 expressing tumor with strongly reduced expression of melanoma differentiation Ags . Persisting tumor-specific Ag expression maintained tumor immune competence suggesting that tumor-specific Ags/melanoma differentiation Ags may complement each other in the context of melanoma Ag-specific vaccination . DB00741 is a suppressor of apoptosis in bovine corpus luteum . Glucocorticoid ( GC ) acts as a modulator of physiological functions in several organs . In the present study , we examined whether GC suppresses luteolysis in bovine corpus luteum ( CL ) . DB00741 ( an active GC ) reduced the mRNA expression of caspase 8 ( Q14790 ) and caspase 3 ( P42574 ) and reduced the enzymatic activity of P42574 and cell death induced by tumor necrosis factor ( P01375 ) and interferon gamma ( P01579 ) in cultured bovine luteal cells . mRNAs and proteins of GC receptor ( P04150 ) , 11beta-hydroxysteroid dehydrogenase type 1 ( P28845 ) , and P80365 were expressed in CL throughout the estrous cycle . Moreover , the protein expression and the enzymatic activity of P28845 were high at the early and the midluteal stages compared to the regressed luteal stage . These results suggest that cortisol suppresses P01375 - P01579 -induced apoptosis in vitro by reducing apoptosis signals via Q14790 and P42574 in bovine CL and that the local increase in cortisol production resulting from increased P28845 at the early and midluteal stages helps to maintain CL function by suppressing apoptosis of luteal cells . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Chemical synthesis and in vitro biological evaluation of a phosphorylated bisubstrate inhibitor of type 3 17beta-hydroxysteroid dehydrogenase . Type 3 17beta-hydroxysteroid dehydrogenase ( 17beta-HSD ) catalyzes the last step in the biosynthesis of the potent androgen testosterone ( T ) by selectively reducing the C17 ketone of 4-androstene-3,17-dione ( delta4-dione ) , with NADPH as cofactor . This enzyme is thus an interesting therapeutic target for androgen-sensitive diseases . Using an efficient convergent chemical approach we synthesized a phosphorylated version of the best delta4-dione/adenosine hybrid inhibitor of type 3 17beta-HSD previously reported . An appropriately protected P06681 ' phosphorylated adenosine was first prepared and linked by esterification to the steroid delta4-dione bearing an alkyl spacer . After three deprotection steps , the phosphorylated bisubstrate inhibitor was obtained . The inhibitory potency of this compound was evaluated on homogenated P29320 -293 cells overexpressing type 3 17beta-HSD and compared to the best non-phosphorylated bisubstrate inhibitor . Unexpectedly , the phosphorylated derivative was slightly less potent than the non-phosphorylated bisubstrate inhibitor of type 3 17beta-HSD . Two hypotheses are discussed to explain this result : 1 ) the phosphorylated adenosine moiety does not interact optimally with the cofactor-binding site and 2 ) the bisubstrate inhibitors , phosphorylated or not , interact only with the substrate-binding site of type 3 17beta-HSD . The anti-proliferative effects of DB00136 on breast and prostate cancer cells are associated with induction of P38398 gene expression . The anti-proliferative action of the seco-steroid hormone 1alpha , 25-dihydroxyvitamin D3 [ DB00136 ] extends to some , but not all breast and prostate cancer cell lines . By elucidating the molecular mechanisms mediating the sensitivity of these cells , we can identify critical target genes regulated directly or indirectly by DB00136 and pathways potentially disrupted during transformation . In this study , we demonstrated the induction of expression of P38398 mRNA and protein as well as transcriptional activation from the P38398 -promoter by DB00136 in the sensitive breast cancer cell line MCF-7 . This was not observed in the DB00136 -resistant breast cancer cell line MDA-MB-436 . The induction of P38398 mRNA was blocked by cyclohexamide . This indicated that transcriptional activation was mediated indirectly by the vitamin D receptor ( P11473 ) . Inhibition of P11473 protein levels by stable transformation of the anti-sense P11473 in MCF-7 reduced the sensitivity of MCF-7 to DB00136 by 50-fold . In addition , the induction of P38398 protein and transcriptional activation of a P38398 promoter-luciferase reporter construct was abrogated in the stable transformant with the greatest reduction of P11473 levels . Examination of other breast and prostate cancer cell lines revealed that sensitivity to the anti-proliferative effects of 1alpha , 25(OH)2D3 was strongly associated with an ability to modulate P38398 protein . Furthermore , the expression of the estrogen receptor in these cell lines strongly correlated with their sensitivity to DB00136 and their ability to modulate P38398 expression . Taken together , our data support a model whereby the anti-proliferative effects of DB00136 are mediated , in part , by the induction of P38398 gene expression via transcriptional activation by factors induced by the P11473 and that this pathway is disrupted during the development of prostate and breast cancers . Lack of association between genetic variation in 9 innate immunity genes and baseline CRP levels . It is well-known that baseline levels of P02741 ( CRP ) are an independent cardiovascular risk factor . We hypothesized that genetic variation with significant influence on CRP levels might be found in genes of the innate immunity system . We performed a candidate gene association study examining common single nucleotide polymorphisms in 9 innate immunity genes ( Q9HC29 , P51617 , Q9NWZ3 , P18428 , O95711 , O15553 , O60603 , O00206 and P19838 ) in relation to CRP levels . Seven hundred and seventeen subjects from the Women 's Health Study population were studied : 359 and 358 samples with extremely low ( < 0.2 mg/liter ) and high ( > 5 mg/liter ) CRP levels , respectively . SNPs were identified from publicly available resequencing data , using a minor allele frequency threshold of > 5 % and a linkage disequilibrium ( LD ) -based strategy ( r(2) > 0.8 ) to select 63 LD-independent markers . One non-synonymous SNP in O00206 and two non-synonymous SNPs in Q9HC29 , previously associated with atherosclerosis and Crohn 's disease , respectively , were also studied . Univariate , haplotype and gene-gene interaction analyses all indicated no significant association with CRP levels . Although this work excludes a significant association of common SNPs in these nine genes with CRP levels , it is possible that rarer alleles in these genes , or variation in other innate immunity genes , could be associated with variation in CRP . Effect of the IkBα mutant gene delivery to DB05914 on rat chronic pancreatitis . This study aimed to investigate the effect of inhibitors of the NF-kΒ alpha mutant gene ( IkBaM ) delivery to mensenchymal stem cells ( MSCs ) on rat chronic pancreatitis ( CP ) . A total of 120 Sprague-Dawley rats were randomly divided into 6 groups of 20 : Group A was injected with sterile saline solution , Group B was injected with allogenic MSCs , Group C1 was injected with allogenic IkBαM-MSCs cultured in vitro 4 h before CP modeling , Group P06681 was injected with allogenic IkBαM-MSCs cultured in vitro during CP modeling , Group P01024 was cultured with allogenic IkBαM-MSCs cultured in vitro 4 h after CP modeling , and Group D was injected with rAAV2-MSCs . Cytokine levels of P05362 , P29279 , IL-1 , P05231 , P10145 , P01375 -α , P01033 , P16035 , P22301 , FN , P03956 , P08253 , P08254 , and P14780 were examined . The results indicated that allogenic IκBαM-MSCs could reduce pro-inflammatory cytokine levels and increase anti-inflammatory cytokine levels in CP . The allogenic IkBαM-MSCs reduced the activation and promoted the apoptosis of pancreatic stellate cells in the rat model of CP . IkBαM-MSCs influenced the proliferation and apoptosis of pancreatic stellate cells by regulating the activation of the Q07869 , MAPK , P42345 , TGF-β , NOD-like receptor , Notch , WNT , TGF-β1-SMAD-2/3 , and P04637 signal transduction pathways . Synthesis and biological activities of 14-epi-MART-10 and 14-epi-MART-11 : implications for cancer and osteoporosis treatment . The 14-epimer of MART-10 , namely 14-epi-MART-10 ( 14-epi-2alpha-(3-hydroxypropyl)-1alpha,25-dihydroxy-19-norvitamin D3 ) and its 2-epimeric analog ( 14-epi-MART-11 ) were efficiently synthesized using the Julia coupling reaction to connect between the P01031 and P13671 positions ( steroid numbering ) . An A-ring precursor was prepared from (-)-quinic acid as shown in the previous MART-10 synthesis . The novel 14-epi-CD-ring coupling partner with an elongated two carbon unit as a sulfone was synthesized from 14-epi-25-hydroxy Grundmann 's ketone in good yield . The subsequent coupling reaction followed by a deprotection step afforded a mixture of 14-epi-MART-10 and 14-epi-MART-11 in 40 % yield . To separate 14-epi-MART-10 and 14-epi-MART-11 , each primary hydroxyl group was esterified with a pivaloyl group and the resulting pivalates 2alpha and 2beta were separated by high performance liquid chromatography . After the separation , the P06681 -stereochemistry of each ( 2alpha or 2beta ) was determined by 1H NMR ( nuclear magnetic resonance ) studies including NOE ( nuclear Overhauser effect ) experiments . The pivaloyl group was removed under basic conditions to obtain the target molecules of 14-epi-MART-10 and 14-epi-MART-11 , respectively . The P11473 ( vitamin D receptor ) -binding affinity , HL-60 ( human promyelocytic leukemia ) cell differentiation activity , antiproliferative activity in PZ-HPV-7 ( immortalized normal prostate ) cells and transactivation activity of the osteocalcin promoter in Q9UKB1 ( human osteoblast cell line ) cells ( serum-free conditions ) were investigated . In addition , the effects on bone mineral density ( BMD ) and the blood and urine calcium concentrations of ovariectomized ( OVX ) rats were examined . 14-epi-MART-10 has much greater antiproliferative and cell differentiation activities compared to 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Characterization of a vitamin D receptor knockout mouse as a model of colorectal hyperproliferation and DNA damage . The vitamin D receptor knockout ( P11473 -KO ) mouse presents with a skeletal phenotype typical for complete lack of genomic DB00136 effects . Our previous data from human colorectal tissue suggest that the steroid hormone and its receptor may have protective function against tumour progression . In order to investigate the relevance of the vitamin D system for pre-malignant site-directed changes in the colon , we characterized the amount and site-specific distribution of the P11473 along the large intestine in wild-type ( WT ) , heterozygote ( HT ) and KO mice . We also evaluated expression of proliferating cell nuclear antigen ( P12004 ) , of cyclin D1 and the levels of 8-hydroxy-2'-deoxyguanosine ( 8-OHdG ) , a marker of oxidative stress . In colon ascendens , proliferative cells were dispersed all along the crypt and expression levels of all three markers were high in WT mice . A decrease of P11473 expression did not affect expression significantly . In colon descendens , however , fewer proliferative cells were solely located in the lower third of the crypt , and an inverse relationship between P11473 reduction , P12004 positivity and cyclin D1 expression was found in HT and KO mice . In parallel to enhanced proliferation a highly significant increase of 8-OHdG positivity occurred . Therefore , the sigmoid colon of P11473 -KO mice , fed on an appropriate lactose/calcium-enriched diet to alleviate impaired calcium homeostasis-related phenotypic changes , is an excellent model for investigating induction and prevention of pre-malignant changes in one of the hotspots for human colorectal cancer incidence . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Q15109 /RAGE-mediated autophagy promotes pancreatic tumorigenesis and bioenergetics through the P05231 -pSTAT3 pathway . Pancreatic ductal adenocarcinoma ( PDA ) , the fourth leading cause of cancer death in the United States , is a complex disease that arises in the setting of genetic alterations ( P01116 , P38398 , Q13485 , CDKN2A/p16 ( INK4a ) and P04637 ) , epigenetic perturbations ( MIR155 , acetylation and methylation ) and epicellular events ( diabetes and inflammation ) . We have demonstrated that the advanced glycation end product-specific receptor ( Q15109 , also called RAGE ) contributes to pancreatic tumorigenesis . Targeted ablation of Q15109 diminishes the amount of autophagic flux and attenuates the development of early pancreatic intraepithelial neoplasia ( PanIN ) lesions in a murine model of P01116 -drivien carcinogenesis . Autophagy ( programmed cell survival ) , a metabolic process of lysosome-mediated self-digestion , promotes pancreatic cancer growth . In pancreatic tumor cell lines , Q15109 -mediated autophagy promotes interleukin-6 ( P05231 ) -induced phosphorylation of signal transducer and activator of transcription 3 ( pSTAT3 ) and mitochondrial localization of pSTAT3 . Enhanced mitochondrial pSTAT3 increases the pool of available DB00171 and increases cellular proliferation . Moreover , we observed a positive feedback loop between activation of autophagy and the P05231 -pSTAT3 pathway , perhaps different from the role of cytosolic nonphosphorylated P40763 , which has been reported to inhibit autophagy . These Q15109 -dependent changes were found during the earliest stages of pancreatic cancer development . These observations of inflammation and altered metabolism in PDA provide a pathological link to early precursor lesion development . Thus , Q15109 is an important inflammatory mediator that modulates crosstalk between prosurvival pathways , P05231 -pSTAT3 and autophagy , in PDA tumor cells , and contributes to early PanIN formation . Nongenotropic , anti-apoptotic signaling of 1alpha,25(OH)2-vitamin D3 and analogs through the ligand binding domain of the vitamin D receptor in osteoblasts and osteocytes . Mediation by Src , phosphatidylinositol 3- , and JNK kinases . Because sex steroids regulate the life span of bone cells by modulating cytoplasmic kinase activity via a nongenotropic action of their classical receptors , we have explored the possibility that the vitamin D nuclear receptor ( P11473 ) might exhibit similar nongenotropic actions . We report that the conformationally flexible full P11473 agonist , 1alpha,25(OH)2-vitamin D3 ( DB00136 ) , and the 6-s-cis-locked 1alpha,25(OH)2-lumisterol3 ( JN ) analog , also acting through the P11473 but with poor transcriptional activity , protected murine osteoblastic or osteocytic cells from apoptosis . This effect was reproduced in HeLa cells transiently transfected with either wild type P11473 or a mutant consisting of only the P11473 ligand binding domain . The P11473 ligand binding domain bound [3H] DB00136 as effectively as wild type P11473 but did not induce vitamin D response element-mediated transcription . The anti-apoptotic effects of DB00136 and the 6-s-cis-locked 1alpha,25(OH)2-lumisterol3 analog in calvaria cells were blocked by three cytoplasmic kinase inhibitors : Src kinase inhibitor 4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine ( P50391 ) , phosphatidylinositol 3 kinase inhibitor Wortmannin , and the JNK kinase inhibitor SP600125 . However , inhibition of p38 with SB203580 or P29323 with either U0126 or a transfected dominant negative MEK did not interfere with these anti-apoptotic actions . Further , DB00136 induced rapid ( 5 min ) association of P11473 with Src kinase in OB-6 cells . Finally , actinomycin D or cycloheximide prevented the anti-apoptotic effect of DB00136 , indicating that transcriptional events are also required . These findings suggest that nongenotropic modulation of kinase activity is also a general property of the P11473 and that ligands that activate nongenotropic signals , but lack transcriptional activity , display different biological profiles from the steroid hormone DB00136 . Interactions between DB00136 and residues in the ligand-binding pocket of the vitamin D receptor : a correlated fragment molecular orbital study . To provide physicochemical insight into the role of each residue in the ligand-binding pocket ( P18428 ) of the vitamin D receptor ( P11473 ) , we evaluated the energies of the interactions between the P18428 residues and DB00136 by using an ab initio fragment molecular orbital ( FMO ) method at the Møller-Plesset second-order perturbation ( MP2 ) level . This FMO-MP2 method can be used to correctly evaluate both electrostatic and van der Waals dispersion interactions , and it affords these interaction energies separately . We deduced the nature of each interaction and determined the importance of all the P18428 residues involved in ligand recognition by the P11473 . We previously reported the results of alanine-scanning mutational analysis ( ASMA ) of all 34 non-alanine residues lining the P18428 of the human P11473 . The theoretical results in combination with the ASMA results enabled us to assign the role of each P18428 residue . We concluded that electrostatic interactions are the major determinant of the ligand-binding activity and ligand recognition specificity and that van der Waals interactions are important for protein folding and , in turn , for cofactor binding . Nuclear factor of activated T cells ( NFAT ) as a molecular target for 1alpha,25-dihydroxyvitamin D3-mediated effects . The molecular basis of the immunomodulatory properties of 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) remains elusive . We demonstrate here that DB00136 -mediated suppressive effects on the inducible expression of cytokine genes in human T cells may , in part , be due to diminished activity of the transcription factor NFAT . The vitamin D3 receptor ( P11473 ) and its heterodimeric partner retinoid X receptor alpha ( RXR alpha ) specifically bound to the distal NFAT site in the human P60568 promoter , and this binding was abolished by mutating unique regions in the NFAT oligonucleotide . In vitro inhibition of NFAT complex formation was noted when P11473 -RXR alpha heterodimers were added to DNA binding reactions containing nuclear extracts from activated B or T cells , whereas in vitro NFkappaB complex formation was not significantly influenced . Furthermore , DB00136 treatment of activated T cells resulted in decreased formation of NFAT complexes detected upon incubation of nuclear extracts from these cells with 32P-labeled probe . Transient expression of both P11473 and RXR alpha , but not of a single component , was capable of inhibiting expression of a NFAT-driven reporter gene in stimulated jurkat cells in a ligand-dependent manner . These results suggest that NFAT plays a crucial role in DB00136 -mediated immunosuppressive activity . DB00091 up-regulates Krüppel-like factor-4 ( O43474 ) in vascular smooth muscle cells and drives phenotypic modulation in vivo . DB00091 A ( Q13216 , calcineurin inhibitor ) has been shown to block both vascular smooth muscle cell ( VSMC ) proliferation in cell culture and vessel neointimal formation following injury in vivo . The purpose of this study was to determine molecular and pathological effects of Q13216 on VSMCs . Using real-time reverse transcription-polymerase chain reaction , Western blot analysis , and immunofluorescence microscopy , we show that Q13216 up-regulated the expression of Krüppel-like factor-4 ( O43474 ) in VSMCs . O43474 plays a key role in regulating VSMC phenotypic modulation . O43474 antagonizes proliferation , facilitates migration , and down-regulates VSMC differentiation marker gene expression . We show that the VSMC differentiation marker genes smooth muscle alpha-actin ( P62736 ) , transgelin ( Q01995 ) , smoothelin ( P53814 ) , and myocardin ( Q8IZQ8 ) are all down-regulated by Q13216 in VSMC monoculture , whereas cyclin-dependent kinase inhibitor-1A ( P38936 ) and matrix metalloproteinase-3 ( P08254 ) are up-regulated . Q13216 did not affect the abundance of the VSMC microRNA ( MIR ) markers MIR143 and MIR145 . Administration of Q13216 to rat carotid artery in vivo resulted in acute and transient suppression of P62736 , Q01995 , P53814 , Q8IZQ8 , and smooth muscle myosin heavy chain ( P35749 ) mRNA levels . The tumor suppressor genes O43474 , p53 , and P38936 , however , were up-regulated , as well as P08254 , P14780 , and collagen-VIII . Q13216 -treated arteries showed remarkable remodeling , including breakdown of the internal elastic lamina and reorientation of VSMCs , as well as increased O43474 immunostaining in VSMCs and endothelial cells . Altogether , these data show that cyclosporin up-regulates O43474 expression and promotes phenotypic modulation of VSMCs . The human peroxisome proliferator-activated receptor delta gene is a primary target of 1alpha,25-dihydroxyvitamin D3 and its nuclear receptor . Peroxisome proliferator-activated receptor ( Q07869 ) delta is the most widely expressed member of the Q07869 family of nuclear receptor fatty acid sensors . Real-time PCR analysis of breast and prostate cancer cell lines demonstrated that PPARdelta expression was increased 1.5 to 3.2-fold after three hours stimulation with the natural vitamin D receptor ( P11473 ) agonist , 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) . In silico analysis of the 20 kb of the human PPARdelta promoter revealed a Q93038 -type DB00136 response element approximately 350 bp upstream of the transcription start site , which was able to bind P11473 -retinoid X receptor ( RXR ) heterodimers and mediate a DB00136 -dependent upregulation of reporter gene activity . Chromatin immuno-precipitation assays demonstrated that a number of proteins representative for DB00136 -mediated gene activation , such as P11473 , RXR and RNA polymerase II , displayed a DB00136 -dependent association with a region of the proximal PPARdelta promoter that contained the putative Q93038 -type VDRE . This was also true for other proteins that are involved in or are the subject of chromatin modification , such as the histone acetyltransferase CBP and histone 4 , which displayed ligand-dependent association and acetylation , respectively . Finally , real-time PCR analysis demonstrated that DB00136 and the synthetic PPARdelta ligand L783483 show a cell and time-dependent interference in each other 's effects on P11473 mRNA expression , so that their combined application shows complex effects on the induction of P11473 target genes , such as Q07973 . Taken together , we conclude that PPARdelta is a primary DB00136 -responding gene and that P11473 and PPARdelta signaling pathways are interconnected at the level of cross-regulation of their respective transcription factor mRNA levels . Genetic susceptibility to aseptic loosening following total hip arthroplasty : a systematic review . Introduction Aseptic loosening is the most common cause of total hip arthroplasty ( DB00382 ) failure and revision surgery . Genetic polymorphisms could be determinant factors for implant loosening . Source of data We performed a comprehensive search of Medline , CINAHL , Googlescholar , Embase and Cochrane databases , using various combinations of the keyword terms ' aseptic loosening ' , ' gene ' , ' hip arthoplasty ' , ' genetics ' , ' loosening ' . Twelve studies detailing the genetic investigation of patients with aseptic loosening of a DB00382 were identified . Areas of agreement SNPs of GNAS1 , P01375 -238 A allele , P01375 -α promoter ( -308G→A ) transition , P05231 -174 G allele , interleukin ( IL ) -6 ( -597 ) and ( -572 ) , P03956 -promoting gene , C/C genotype for the P03956 , P50281 , P08253 , transforming growth factor-beta1 signal sequence ( 29T→C ) transitions , A/A genotype for the O00300 -163 , and P11226 were overexpressed in patients with aseptic loosening and periprosthetic osteolysis . Areas of controversy Data from single centre studies do not allow one to compare the results of different studies . Conclusion Several gene pathways and genes contribute to the genetic susceptibility to aseptic loosening following DB00382 . Further studies will enhance the understanding of prosthesis failure , and may inform and direct pharmaceutical interventions . Growing points Further multi-centre prospective studies are necessary to confirm the general validity of the findings reported . Single-centre findings should be replicated in other centres and populations to open new avenues for pre-surgical genetic testing and to investigate immune response modulation in DB00382 . Areas timely for developing research Research in this field could lead to better understanding of mechanisms behind aseptic osteolysis , and improve the results of DB00382 . A novel role for farnesyl pyrophosphate synthase in fibroblast growth factor-mediated signal transduction . P14324 ( FPPS ) catalyses the formation of a key cellular intermediate in isoprenoid metabolic pathways . Here we describe a novel role for this enzyme in fibroblast growth factor ( FGF ) -mediated signalling . We demonstrate the binding of FPPS to FGF receptors ( FGFRs ) using the yeast two-hybrid assay , pull-down assays and co-immunoprecipitation . The interaction between FPPS and FGFR is regulated by the cellular metabolic state and by treatment with P09038 . Overexpression of FPPS inhibits P09038 -induced cell proliferation , accompanied by a failure of the P09038 -mediated induction of cyclins D1 and E. Overexpression of FPPS in fibroblasts also promotes increased farnesylation of Ras , and temporally extends P09038 -stimulated activation of the Ras/ P29323 ( extracellular-signal-regulated kinase ) cascade . These data suggest that , in addition to its role in isoprenoid biosynthesis , FPPS may function as a modulator of the cellular response to FGF treatment . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . Inhibition of histamine H1 receptor activity modulates proinflammatory cytokine production of dendritic cells through c-Rel activity . BACKGROUND : DB11320 exerts diverse effects on immune regulation through four types of histamine receptors ( HRs ) . Among them , type 1 receptor ( P35367 ) plays an important role in allergic inflammation . Dendritic cells ( DCs ) , which express at least three types of HRs , are professional antigen-presenting cells controlling the development of allergic inflammation . However , the molecular mechanisms involved in P35367 -mediated NF-ĸB signaling of DCs remain poorly defined . METHODS : Bone-marrow ( BM ) -derived DCs ( BM-DCs ) were treated with P35367 inverse agonists to interrupt basal P35367 -mediated signaling . The crosstalk of P35367 -mediated signaling and the NF-ĸB pathway was examined by NF-ĸB cellular activity using a luciferase reporter assay , NF-ĸB subunit analysis using Western blotting and P01375 -α promoter activity using chromatin immunoprecipitation . RESULTS : Blockage of P35367 signaling by inverse agonists significantly inhibited P01375 -α and P05231 production of BM-DCs . P35367 -specific agonists were able to enhance P01375 -α production , but this overexpression was significantly inhibited by NF-ĸB inhibitor . The P35367 inverse agonist ketotifen also suppressed cellular NF-ĸB activity , suggesting crosstalk between P35367 and NF-ĸB signaling in DCs . After comprehensive analysis of NF-ĸB subunits , c-Rel protein expression was significantly down-regulated in ketotifen-treated BM-DCs , which led to inhibition of the promoter activity of P01375 -α . Finally , adoptive transfer of the ketotifen-treated BM-DCs did not induce significant allergic airway inflammation compared to that of control cells in vivo . CONCLUSIONS : Our results suggest that c-Rel controls P35367 -mediated proinflammatory cytokine production in DCs . This study provides a potential mechanism of P35367 -mediated signaling and NF-ĸB pathway crosstalk in allergic inflammation . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . Hypoxia enhances P61073 expression in human microvascular endothelial cells and human melanoma cells . The influence of environmental factors ( cytokines , matrix components , serum factors and O(2) level ) on expression of receptors for angiogenic versus angiostatic CXC chemokines in human microvascular endothelial cells has not been extensively investigated . Our semi-quantitative RT-PCR analysis demonstrated that P01375 and P01579 repressed P61073 mRNA levels in immortalized human microvascular endothelial HMEC-1 cells after 4 h , whereas only P01375 displayed inhibitory activity in primary human microvascular endothelial cells ( HMVEC ) . P61073 mRNA expression was not affected by P15692 , GM- P04141 , IL-1beta or various basal membrane matrix components , but was significantly up-regulated after serum starvation and/or hypoxic treatment of the microvascular endothelial cells . The alternative P48061 receptor , P25106 /RDC1 , was also up-regulated by hypoxia in HMEC-1 cells , although less consistently than P61073 . Furthermore , hypoxia and serum starvation were required for cell surface display of P61073 and P48061 induction of P29323 activation in HMEC-1 cells . In contrast , P25025 and P49682 mRNA levels remained , respectively , low and undetectable under all the conditions tested , and surface expression of P25025 , P49682 and P25106 on the HMEC- 1 cells could not be demonstrated by FACS . In the human SK- P61006 -5 melanoma cell line , P61073 mRNA expression was also increased under hypoxic conditions , whereas P25025 mRNA levels remained low and levels of P49682 and P25106 were undetectable . However , immunohistochemical staining of human metastatic melanoma sections demonstrated that P25025 , P49682 , P61073 and P25106 are expressed on tumor cells and , to a lesser extent , on endothelial cells . These results demonstrate that the tumor microenvironment regulates chemokine receptor expression through both cytokine and oxygen levels . Metalloproteinases control brain inflammation induced by pertussis toxin in mice overexpressing the chemokine P13500 in the central nervous system . Inflammatory leukocytes infiltrate the CNS parenchyma in neuroinflammation . This involves cellular migration across various structures associated with the blood-brain barrier : the vascular endothelium , the glia limitans , and the perivascular space between them . Leukocytes accumulate spontaneously in the perivascular space in brains of transgenic ( Tg ) mice that overexpress P13500 under control of a CNS-specific promoter . The Tg mice show no clinical symptoms , even though leukocytes have crossed the endothelial basement membrane . Pertussis toxin ( PTx ) given i.p. induced encephalopathy and weight loss in Tg mice . We used flow cytometry , ultra-small superparamagnetic iron oxide-enhanced magnetic resonance imaging , and immunofluorescent staining to show that encephalopathy involved leukocyte migration across the glia limitans into the brain parenchyma , identifying this as the critical step in inducing clinical symptoms . Metalloproteinase ( MPs ) enzymes are implicated in leukocyte infiltration in neuroinflammation . Unmanipulated Tg mice had elevated expression of tissue inhibitor of metalloproteinase-1 , matrix metalloproteinase ( MMP ) -10 , and -12 mRNA in the brain . PTx further induced expression of tissue inhibitor of metalloproteinase-1 , metalloproteinase disintegrins-12 , P22894 , and -10 in brains of Tg mice . Levels of the microglial-associated MP P51511 were not affected in control or PTx-treated Tg mice . PTx also up-regulated expression of proinflammatory cytokines IL-1beta and P01375 mRNA in Tg CNS . Weight loss and parenchymal infiltration , but not perivascular accumulation , were significantly inhibited by the broad-spectrum MP inhibitor BB-94/ DB03880 . Our finding that MPs mediate PTx-induced parenchymal infiltration to the chemokine-overexpressing CNS has relevance for the pathogenesis of human diseases involving CNS inflammation , such as multiple sclerosis . Q13507 -like protein and vitamin D receptor mediate DB00136 -induced Q5T124 influx in muscle cells . 1alpha,25-Dihydroxy-Vitamin-D3 ( 1alpha,25(OH)2- DB00169 ) stimulates in skeletal muscle cells Ca2+ release from inner stores and influx through both voltage-dependent and store-operated Ca2+ ( Q5T124 , CCE ) channels . We investigated the involvement of TRPC proteins and Vitamin D receptor ( P11473 ) in CCE induced by DB00136 in chick muscle cells . Two fragments were amplified by RT-PCR , exhibiting approximately 80 % sequence homology with mammalian Q13507 /6/7 . Northern and Western blots employing a Q13507 -probe and anti- Q13507 antibodies , respectively , confirmed endogenous expression of a Q13507 -like protein of 140 kDa . Spectrofluorimetric measurements in Fura-2 loaded cells showed reduced CCE and Mn2+ entry in response to either thapsigargin or DB00136 upon transfection with anti- Q13507 /6/7 antisense oligodeoxynucleotides ( ODNs ) . Transfection with anti- P11473 antisense ODNs diminished DB00136 -dependent Ca2+ and Mn2+ influx . Co-immunoprecipitation of Q13507 -like protein and P11473 under non-denaturating conditions was observed . We propose that endogenous Q13507 -like proteins and the P11473 participate in the modulation of CCE by DB00136 in muscle cells , which could be mediated by an interaction between these proteins . The genes of the coactivator Q15596 and the corepressor Q9Y618 are primary DB00136 targets . The complex of the receptor for the hormone 1alpha,25-dihydroxyvitamin D(3) ( 1alpha,25(OH)(2)D(3) ) , Vitamin D(3) receptor ( P11473 ) , the retinoid X receptor ( RXR ) and a 1alpha,25(OH)(2)D(3) response element ( VDRE ) is considered to be the molecular switch for nuclear 1alpha,25(OH)(2)D(3) signaling . In the presence of ligand the P11473 -RXR complex interacts with coactivator ( DB01992 ) proteins that in turn contact components of the basal transcriptional machinery resulting in an enhanced transcription of 1alpha,25(OH)(2)D(3) target genes . In the absence of ligand the P11473 remains bound to the DNA and interacts with corepressor ( CoR ) proteins that are involved in gene silencing activity . We treated MCF-7 breast cancer cells with 1alpha,25(OH)(2)D(3) for increasing amounts of time , extracted mRNA and screened by real-time PCR the members of the P52701 DB01992 and NCoR CoR families . We find that of the P52701 coactivators , only Q15596 was responsive to 1alpha,25(OH)(2)D(3) . Similarly Q9Y618 but not NCoR1 gene transcription was sensitive to 1alpha,25(OH)(2)D(3) treatment . In silico analysis revealed that both Q15596 and Q9Y618 promoters have substantial numbers of VDREs compared to the promoters of the other family members . These VDREs are formed by direct repeats of the core binding motif RGKTCA with a three nucleotide spacing ( Q93038 ) . We suggest that some or all of these Q93038 -type VDREs are responsible for the observed responsiveness of Q15596 and Q9Y618 to 1alpha,25(OH)(2)D(3) . Expression and effect of fibroblast growth factor 9 in bovine theca cells . P31371 ( P31371 ) protein affects granulosa cell ( GC ) function but is mostly localized to theca cell ( TC ) and stromal cell of rat ovaries . The objectives of this study were to determine the 1 ) effects of P31371 on TC steroidogenesis , gene expression , and cell proliferation ; 2 ) mechanism of action of P31371 on TCs ; and 3 ) hormonal control of P31371 mRNA expression in TCs . Bovine ovaries were collected from a local slaughterhouse and TCs were collected from large ( 8-22 mm ) follicles and treated with various hormones in serum-free medium for 24 or 48 h . P31371 caused a dose-dependent inhibition ( P < 0·05 ) of LH- and LH+IGF1-induced androstenedione and progesterone production . Also , P31371 inhibited ( P < 0·05 ) LH+IGF1-induced expression of P22888 , P05108 , and P05093 mRNA ( via real-time RT-PCR ) in TCs . P31371 had no effect ( P > 0·10 ) on STAR mRNA abundance . Furthermore , P31371 inhibited dibutyryl DB02527 -induced progesterone and androstenedione production in LH+IGF1-treated TCs . By contrast , P31371 increased ( P < 0·05 ) the number of bovine TCs . Abundance of P31371 mRNA in GCs and TCs was several-fold greater ( P < 0·05 ) in small ( 1-5 mm ) vs large follicles . P01375 α and P41221 increased ( P < 0·05 ) abundance of P31371 mRNA in TCs . In summary , expression of P31371 mRNA in TCs is developmentally and hormonally regulated . P31371 may act as an autocrine regulator of ovarian function in cattle by slowing TC differentiation via inhibiting LH+IGF1 action via decreasing gonadotropin receptors and the DB02527 signaling cascade while stimulating proliferation of TCs . Role of transient receptor potential P01024 in P01375 -enhanced calcium influx in human airway myocytes . Previous studies have suggested that the proinflammatory cytokine , P01375 , contributes to airway hyperresponsivness by altering airway smooth muscle ( P17405 ) Ca(2+) responses to agonist stimulation . The present study examined the effects of P01375 on Ca(2+) influx pathways in cultured human P17405 cells ( HASMCs ) . Proteins encoded by the transient receptor potential ( TRP ) gene family function as channels through which receptor-operated and store-operated Ca(2+) entry ( SOCE ) occur . In the present study , the presence of P48995 , Q13507 , Q9UBN4 , Q9UL62 , and Q9Y210 mRNA and protein expression was confirmed in cultured HASMCs using RT-PCR and Western blot analysis . P01375 treatment significantly increased Q13507 mRNA and protein levels in HASMCs as well as SOCE . P01375 treatment also increased both the peak and plateau intracellular Ca(2+) concentration responses in HASMCs elicited by acetylcholine and bradykinin . The effects of P01375 treatment on SOCE and agonist-induced intracellular Ca(2+) concentration responses were attenuated using small interfering RNA transfection , which knocked down Q13507 expression . Thus , in inflammatory airway diseases , P01375 treatment may result in increased myocyte activation due to altered Ca(2+) influx pathways . These results suggest that Q13507 may be an important therapeutic target in inflammatory airway diseases such as asthma and chronic obstructive pulmonary disease . Orai proteins interact with TRPC channels and confer responsiveness to store depletion . The TRPC ( C-type transient receptor potential ) class of ion channels has been hypothesized to participate in store-operated Ca(2+) entry ( SOCE ) . Recently , however , Q13586 and Orai1 proteins have been proposed to form SOCE channels . Whether TRPCs participate in SOCE that is dependent on or regulated by Orai has not been explored . Here we show that Orai1 physically interacts with the N and C termini of Q13507 and Q9Y210 , and that in cells overexpressing either Q13507 or Q9Y210 in a store-depletion insensitive manner , these TRPCs become sensitive to store depletion upon expression of an exogenous Orai . Thus , Orai-1 , -2 , and -3 enhanced thapsigargin-induced calcium entry by 50-150 % in cells stably overexpressing either Q13507 or Q9Y210 . Orai1 expression had no significant effect on endogenous , thapsigargin-induced calcium entry in wild-type cells ( P29320 -293 , COS1 ) , in P29320 cells expressing a thapsigargin-sensitive variant of Q13507 ( TRPC3a ) , or in P29320 cells overexpressing another membrane protein , P37288 . Single-channel cation currents present in membrane patches of Q13507 -overexpressing cells were suppressed by expression of Orai1 . We propose that Orai proteins by interacting with TRPCs act as regulatory subunits that confer Q13586 -mediated store depletion sensitivity to these channels . P18075 blocks the cyclosporine-A-induced epithelial-to-mesenchymal transition in renal tubular epithelial cells . AIMS : The nephrotoxic side effects of cyclosporine A ( DB00091 ) are partly due to induction of the epithelial-to-mesenchymal transition ( EMT ) , possibly through upregulation of connective tissue growth factor ( P29279 ) . Bone morphogenetic protein ( P18075 ) has been reported to protect against and reverse renal injury via its renotropic and antifibrotic effects . We therefore designed a method to investigate the mechanism by which P18075 inhibits Q13216 -induced EMT in cultured human renal proximal tubular epithelial ( HK-2 ) cells and whether P18075 can antagonize DB00091 -induced profibrogenic effects . METHODS : Cultured HK-2 cells were treated with DB00091 or a combination of DB00091 and P18075 for 72 h . Morphological changes were assessed by phase-contrast microscopy . The expression of alpha-smooth muscle actin ( alpha-SMA ) , P12830 , collagen type I and P29279 was analyzed by immunofluorescence , RT-PCR , and Western blot . Additionally , the effect of P29279 silencing on DB00091 -mediated gene expression was assessed . RESULTS : DB00091 -induced EMT was associated with decreased expression of P12830 , increased expression of alpha-SMA , collagen type I and P29279 , and loss of epithelial morphology . P18075 inhibited these effects in a dose-dependent manner . Using siRNA , P29279 knock-down also attenuated EMT-associated phenotypic changes induced by DB00091 . CONCLUSION : These data suggest that P18075 can block DB00091 -induced EMT by downregulating the expression of P29279 , a downstream mediator of EMT . An in vitro osteoclast-forming assay to measure myeloma cell-derived osteoclast-activating factors . Much of the morbidity and mortality associated with the plasma cell ( PC ) malignancy , multiple myeloma ( MM ) , is owing to the severe osteolytic bone disease seen in patients with this disease . Although the molecular mechanisms responsible for osteolysis remain to be fully elucidated , it is clear from numerous studies that it is owing , in part , to an increase in osteoclastic bone resorption . Several known osteoclast ( OC ) -activating factors ( OAFs ) are produced by myeloma PCs ( MPCs ) , or by stromal cells in response to MPCs and include interleukin-1beta ( IL-1beta ) ; tumor necrosis factor-alpha ( P01375 ) ; P05231 ; parathyroid hormone-related protein ; macrophage inflammatory protein-1alpha ; and , most recently , the P01375 -ligand family member receptor activator of nuclear factor-kappaB ligand ( O14788 ) . The identification and significance of any one of these myeloma-derived OAFs is dependent on robust and reliable assays that measure the de novo formation and activation of OCs . A number of in vitro assay systems have been described that examine the requirements for normal OC formation and are easily adaptable for examining which MM-derived Q86UD1 and to what extent it is responsible for the bone loss observed in individuals with myeloma . This chapter describes one such in vitro model system . P12004 D-1 , interleukin-6 , HER-2/neu , transforming growth factor receptor-II and prediction of relapse in women with early stage , hormone receptor-positive breast cancer treated with tamoxifen . We hypothesized that amplification or overexpression of HER-2 ( c-erbB-2 ) , the Ki-67 antigen ( Mib1 ) , cyclin D-1 ( CD1 ) , interleukin-6 ( P05231 ) , or the transforming growth factor beta II receptor , ( TGFbetaRII ) , would predict relapse in women with early stage , estrogen ( ER ) and/or progesterone receptor ( PR ) positive breast cancer treated with tamoxifen . Conditional logistic regression models and a new novel analytic method - support vector machines ( SVM ) were used to assess the effect of multiple variables on treatment outcome . All patients had stage I-IIIa breast cancer ( AJCC version 5 ) . We paired 63 patients who were disease-free on or after tamoxifen with 63 patients who had relapsed ( total 126 ) ; both disease-free and relapsed patients were matched by duration of tamoxifen therapy and time to recurrence . These 126 patients also served as the training set for SVM analysis and 18 other patients used as a validation set for SVM . In a multivariate analysis , larger tumor size , increasing extent of lymph node involvement , and poorer tumor grade were significant predictors of relapse . When HER-2 or CD1 were added to the model both were borderline significant predictors of relapse . The SVM model , after including all of the clinical and marker variables in the 126 patients as a training set , correctly predicted relapse in 78 % of the 18 patient validation samples . In this trial , HER-2 and CD1 proved of borderline significance as predictive factors for recurrence on tamoxifen . An SVM model that included all clinical and biologic variables correctly predicted relapse in > 75 % of patients . Increased serum osteoprotegerin in patients with primary adrenal insufficiency receiving conventional hydrocortisone substitution . Patients treated for primary adrenal insufficiency ( P05121 ) are at risk of steroid over-replacement , which may affect their skeleton . The study was aimed to investigate the effect of steroid substitution on serum osteoprotegerin and receptor activator of nuclear factor kappa-beta ligand ( O14788 ) levels in relation to bone mineral density ( BMD ) in P05121 . Eighty patients ( mean age 47.2±14.5 years , mean hydrocortisone dose 0.49±0.14 mg/kg/day ) and 63 healthy subjects were included . Serum osteoprotegerin , O14788 , 25-hydroxyvitamin D₃ , calcium , phosphate , alkaline phosphatase , intact parathormone , and dehydroepiandrosterone-sulfate levels were evaluated in patients and controls . BMD was assessed in affected subjects using dual-energy X-ray absorptiometry . Mean osteoprotegerin concentration in P05121 patients appeared significantly higher vs. controls ( p=0.002 ) , while O14788 levels were similar ( p=0.430 ) . Serum osteoprotegerin increased with age ( p < 0.001 ) , but showed no correlation with daily hydrocortisone dose . O00300 was negatively correlated with serum dehydroepiandrosterone-sulfate ( p=0.008 ) and with BMD at the lumbar spine ( p < 0.001 ) and femoral neck ( p=0.003 ) . O14788 correlated negatively with P05121 duration ( p=0.029 ) and positively with daily hydrocortisone dose ( p=0.018 ) . Lumbar spine osteoporosis and osteopenia were found in 12 and 31 patients , respectively , whereas in femoral neck : in 5 and 33 individuals . Patients with osteoporosis displayed higher osteoprotegerin levels , but after the age-adjustment the correlation was lost . In conclusion , increased osteoprotegerin in P05121 might reflect a compensatory response to enhanced bone resorption due to exogenous steroid excess and/or result from a deficit in adrenal androgens . O14788 levels remain within normal range on standard steroid replacement . Polymorphisms in genes implicated in dopamine , serotonin and noradrenalin metabolism suggest association with cerebrospinal fluid monoamine metabolite concentrations in psychosis . BACKGROUND : Homovanillic acid ( HVA ) , 5-hydroxyindoleacetic acid ( 5-HIAA ) and 3-methoxy-4-hydroxyphenylglycol ( MHPG ) are the major monoamine metabolites in the central nervous system ( CNS ) . Their cerebrospinal fluid ( P04141 ) concentrations , reflecting the monoamine turnover rates in CNS , are partially under genetic influence and have been associated with schizophrenia . We have hypothesized that P04141 monoamine metabolite concentrations represent intermediate steps between single nucleotide polymorphisms ( SNPs ) in genes implicated in monoaminergic pathways and psychosis . METHODS : We have searched for association between 119 SNPs in genes implicated in monoaminergic pathways [ tryptophan hydroxylase 1 ( P17752 ) , Q8IWU9 , tyrosine hydroxylase ( TH ) , P20711 ( DDC ) , dopamine beta-hydroxylase ( P09172 ) , catechol-O-methyltransferase ( P21964 ) , monoamine oxidase A ( P21397 ) and P27338 ] and monoamine metabolite concentrations in P04141 in 74 patients with psychotic disorder . RESULTS : There were 42 nominally significant associations between SNPs and P04141 monoamine metabolite concentrations , which exceeded the expected number ( 20 ) of nominal associations given the total number of tests performed . The strongest association ( p = 0.0004 ) was found between P27338 rs5905512 , a SNP previously reported to be associated with schizophrenia in men , and MHPG concentrations in men with psychotic disorder . Further analyses in 111 healthy individuals revealed that 41 of the 42 nominal associations were restricted to patients with psychosis and were absent in healthy controls . CONCLUSIONS : The present study suggests that altered monoamine turnover rates in CNS reflect intermediate steps in the associations between SNPs and psychosis . Osteoclast progenitors from cats with and without tooth resorption respond differently to 1,25-dihydroxyvitamin D and interleukin-6 . Both vitamin D and inflammatory cytokines can stimulate osteoclast formation and activity . We studied the effect of DB00136 ( 1,25(OH)(2)D ) , and interleukin-6 ( P05231 ) , on the formation and activity of feline osteoclasts , using peripheral blood mononuclear cells ( PBMCs ) from cats with and without tooth resorption ( TR(+) and TR(-) ) as a source of osteoclast precursors . The formation of osteoclast-like cells ( defined as multinucleated , tartrate-resistant acid phosphatase-positive cells ) was assessed at 7 and 14 days . In the presence of P09603 and O14788 , with and without P05231 , more osteoclasts were formed from TR(-) PBMCs than from TR(+) PBMCs on plastic . More osteoclasts were formed from TR(+) PBMCs on bone slices in the presence of P09603 / O14788 with 1,25(OH)(2)D . This opposite effect may be due to a higher expression of the vitamin D receptor ( P11473 ) in TR(+) osteoclasts and precursors on bone . Formation of resorption pits was analyzed and confirmed with scanning electron microscopy . In conclusion , we propose that TR(+) PBMCs when cultured on bone are sensitive to 1,25(OH)(2)D , whereas the differentiation of TR(-) PMBCs on bone seem more sensitive to P05231 , suggesting that osteoclast precursors from cats with and without tooth resorption respond differently to osteoclast stimulating factors . 1,25-Dihydroxycholecalciferol enhances butyrate-induced P38936 (Waf1/Cip1) expression . Butyrate , a short-chain fatty acid produced in the colon , as well as its prodrug tributyrin , reduce proliferation and increase differentiation of colon cancer cells . P38936 (Waf1/Cip1) and p27(Kip1) are negative regulators of cell cycle and are thought to have a key function in the differentiation of various cell lines . We studied the effects of butyrate on differentiation , P11473 expression , as well as on P38936 (Waf1/Cip1) and p27(Kip1) expression in human colon cancer cells ( Caco-2 ) . Butyrate induced cell differentiation , which was further enhanced after addition of DB00136 . Synergistic effect of butyrate and dihydroxycholecalciferol in Caco-2 cells was due to butyrate-induced overexpression of P11473 . While butyrate as well as dihydroxycholecalciferol increased P38936 (Waf1/Cip1) and p27(Kip1) expression , in contrast combined exposure of butyrate and dihydroxycholecalciferol resulted in a synergistic amplification of P38936 (Waf1/Cip1) , but not of p27(Kip1) expression . These data imply that butyrate selectively increases P38936 (Waf1/Cip1) expression via upregulation of P11473 in Caco-2 cells . Inhibition by 1alpha,25-dihydroxyvitamin D3 of activin A-induced differentiation of murine erythroleukemic P12259 -5 cells . DB00136 ( 1alpha,25-(OH)2D3 ) and other vitamin D3 ( VD3 ) analogs enhanced the inhibitory effect of Activin A on murine erythroleukemia ( P61006 ) cell proliferation and differentiation in a dose-dependent manner . 1alpha,25-(OH)2D3 inhibited differentiation more potently than proliferation by one order of magnitude . The VD3 analog study demonstrated either effect of VD3 on P61006 cells via vitamin D receptor ( P11473 ) , as evidenced from the close relationship with the reported affinities for P11473 . The effects of 1alpha,25-(OH)2D3 were preceded by the suppression of ornithine decarboxylase ( ODC ) activity , a rate-limiting enzyme in polyamine metabolism . Difluoromethylornithine ( DB06243 ) , an inhibitor of ODC , inhibited P61006 cell proliferation , which was reversed by the simultaneous addition of putrescine , a product of ODC , but did not affect differentiation . 1alpha,25-(OH)2D3 inhibited cell differentiation during the phenotype-expression stage as reflected by the inhibition of beta-globin gene expression , while it inhibited proliferation in the commitment stage . Furthermore , it seems unlikely that the different effects of VD3 on proliferation and differentiation may be a result of upregulation of P11473 or nongenomic action . In summary , it was suggested that 1alpha,25-(OH)2D3 inhibited Activin A-induced P61006 cell proliferation and differentiation by distinct mechanisms and inhibited the proliferation by inhibiting ODC activity . We demonstrated the presence of 1alpha,25-(OH)2D3 action on leukemic cells at physiological concentration , which was distinct from the pharmacological effect of VD3 reported thus far . Nuclear translocation of Q02750 triggers a complex T cell response through the corepressor silencing mediator of retinoid and thyroid hormone receptor . Q02750 phosphorylates P27361 /2 and regulates T cell generation , differentiation , and function . Q02750 has recently been shown to translocate to the nucleus . Its nuclear function is largely unknown . By studying human P01730 T cells , we demonstrate that a low level of Q02750 is present in the nucleus of P01730 T cells under basal conditions . T cell activation further increases the nuclear translocation of Q02750 . Q02750 interacts with the nuclear receptor corepressor silencing mediator of retinoid and thyroid hormone receptor ( Q9Y618 ) . Q02750 reduces the nuclear level of Q9Y618 in an activation-dependent manner . Q02750 is recruited to the promoter of c-Fos upon TCR stimulation . Conversely , Q9Y618 is bound to the c-Fos promoter under basal conditions and is removed upon TCR stimulation . We examined the role of Q9Y618 in regulation of T cell function . Small interfering RNA-mediated knockdown of Q9Y618 results in a biphasic effect on cytokine production . The production of the cytokines P60568 , P05112 , P22301 , and IFN-γ increases in the early phase ( 8 h ) and then decreases in the late phase ( 48 h ) . The late-phase decrease is associated with inhibition of T cell proliferation . The late-phase inhibition of T cell activation is , in part , mediated by P22301 that is produced in the early phase and , in part , by β-catenin signaling . Thus , we have identified a novel nuclear function of Q02750 . Q02750 triggers a complex pattern of early T cell activation , followed by a late inhibition through its interaction with Q9Y618 . This biphasic dual effect most likely reflects a homeostatic regulation of T cell function by Q02750 . P12259 Q506 mutation and the low prevalence of cardiovascular disease in Canadian Inuit . BACKGROUND : The Keewatin Inuit have a very low age-adjusted mortality rate from vascular diseases compared to the general population of Canada . Interethnic differences in both genetic and lifestyle factors have been offered as explanations for this observation . We previously found that compared to white subjects , the Inuit had a significantly increased prevalence of 4 candidate alleles for atherosclerosis-related phenotypes , namely AGT T235 , P12104 Q92917 , PONI R192 , and P02649 E4 , and a significantly decreased prevalence of 2 candidate alleles for atherosclerosis-related phenotypes , namely P12821 D and P42898 677T . METHODS : We tested the hypothesis that 165 Canadian Inuit would have a significantly different frequency of the thrombosis-associated P12259 Q506 allele compared with reference controls . RESULTS : We found a complete absence of P12259 Q506 in 165 Inuit , which was significantly different from the frequency of 3.92 % observed in regional control White subjects . CONCLUSIONS : The aggregate of results from our studies in Inuit to date suggests that the beneficial influence of the low prevalence of any or all of the P12821 D , P42898 677T and P12259 Q506 outweighs the deleterious influence of the high prevalence of any or all of the other disease-associated alleles . However , it remains possible that other genetic and/or environmental factors determine the low susceptibility to vascular disease in the Inuit . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Vitamin D regulates the gut microbiome and protects mice from dextran sodium sulfate-induced colitis . The active form of vitamin D [ DB00136 , 1,25(OH)2D3 ] and the vitamin D receptor ( P11473 ) regulate susceptibility to experimental colitis . The effect of the bacterial microflora on the susceptibility of C57BL/6 mice to dextran sodium sulfate-induced colitis was determined . Mice that can not produce 1,25(OH)2D3 [ Cyp27b1 ( Cyp ) knockout ( KO ) ] , P11473 KO as well as their wild-type littermates were used . Cyp KO and P11473 KO mice had more bacteria from the Bacteroidetes and Proteobacteria phyla and fewer bacteria from the Firmicutes and Deferribacteres phyla in the feces compared with wild-type . In particular , there were more beneficial bacteria , including the Lactobacillaceae and Lachnospiraceae families , in feces from Cyp KO and P11473 KO mice than in feces from wild-type . Helicobacteraceae family member numbers were elevated in Cyp KO compared with wild-type mice . Depletion of the gut bacterial flora using antibiotics protected mice from colitis . 1,25(OH)2D3 treatment ( 1.25 μg/100 g diet ) of Cyp KO mice decreased colitis severity and reduced the numbers of Helicobacteraceae in the feces compared with the numbers in the feces of untreated Cyp KO mice . The mechanisms by which the dysbiosis occurs in P11473 KO and Cyp KO mice included lower expression of P12830 on gut epithelial and immune cells and fewer tolerogenic dendritic cells that resulted in more gut inflammation in P11473 and Cyp KO mice compared with wild-type mice . Increased host inflammation has been shown to provide pathogens with substrates to out-compete more beneficial bacterial species . Our data demonstrate that vitamin D regulates the gut microbiome and that 1,25(OH)2D3 or P11473 deficiency results in dysbiosis , leading to greater susceptibility to injury in the gut . Microarray analysis revealed different gene expression patterns in HepG2 cells treated with low and high concentrations of the extracts of Anacardium occidentale shoots . In this study , the effects of low and high concentrations of the Anacardium occidentale shoot extracts on gene expression in liver HepG2 cells were investigated . From MTT assays , the concentration of the shoot extracts that maintained 50 % cell viability ( IC(50) ) was 1.7 mg/ml . Cell viability was kept above 90 % at both 0.4 mg/ml and 0.6 mg/ml of the extracts . The three concentrations were subsequently used for the gene expression analysis using Affymetrix Human Genome 1.0 S.T arrays . The microarray data were validated using real-time qRT-PCR . A total of 246 , 696 and 4503 genes were significantly regulated ( P < 0.01 ) by at least 1.5-fold in response to 0.4 , 0.6 and 1.7 mg/ml of the extracts , respectively . Mutually regulated genes in response to the three concentrations included CDKN3 , LOC100289612 , P00374 , Q99986 , Q99741 , Q96GD4 and P78334 . Genes like Q07973 , P38398 , O14965 , P06493 , P24941 , P11802 and P06213 were significantly regulated at 0.6 mg/ml and 1.7 mg but not at 0.4 mg/ml . However , the expression of genes including O75473 , P17936 , P06400 , P14735 , P01130 , P55157 , P04114 , MTIX , P04179 and P08294 were exclusively regulated at the IC(50) concentration . In conclusion , low concentrations of the extracts were able to significantly regulate a sizable number of genes . The type of genes that were expressed was highly dependent on the concentration of the extracts used . DB01373 as a mediator between erythropoietin and protein tyrosine phosphatase 1B . Erythropoietin ( Epo ) is crucial for promoting the survival , proliferation , and differentiation of mammalian erythroid progenitors . The central role played by tyrosine phosphorylation of erythropoietin receptor ( EpoR ) in Epo-cell activation has focused attention on protein tyrosine phosphatases ( PTPs ) as candidates implicated in the pathogenesis of the resistance to therapy with human recombinant Epo . Prototypic member of the PTP family is P18031 , which has been implicated in the regulation of EpoR signaling pathways . In previous reports we have shown that P18031 is reciprocally modulated by Epo in undifferentiated UT-7 cell line . However , no information is available with respect to the modulation of this phosphatase in non-Epo depending cells or at late stages of erythroid differentiation . In order to investigate these issues we induced UT-7 cells to differentiate and studied their P18031 expression pattern . Simultaneous observations were performed in TF-1 cells which can be cultured either with GM- P04141 , P08700 or Epo . We found that Epo induced P18031 cleaveage in TF-1 and differentiated UT-7 cells . This pattern of P18031 modulation may be due to an increased Q13507 / Q9Y210 expression ratio which could explain the larger and sustained calcium response to Epo and calpain activation in Epo treated TF-1 and differentiated UT-7 cells . Association of novel SNPs in the candidate genes affecting caprine milk fatty acids related to human health . In the present investigation , 618 milk samples of Sirohi breed of goat were collected , and analyzed for conjugated linoleic acid ( DB01211 , C18:2 ) and other fatty acids . The DB01211 in studied goat milk samples was 4.87 mg/g of milk fat and C18:2 cis-9 , trans-11 contributes 2.9 mg/g of milk fat and trans10 cis12 contributes 0.82 mg/g of milk fat . The saturated fatty acids in the milk accounted for 69.55 % and unsaturated fatty acid accounted for 28.50 % . The unsaturated fatty acid was constituted by monounsaturated fatty acid ( 24.57 % ) and polyunsaturated fatty acids ( 3.96 % . ) . The major contribution ( 45.56 % ) in total fatty acid was of C12:0 , C14:0 and C16:0 . C18:0 and short chain ones ( C4:0 , P13671 :0 , Q99618 :0 , and Q99622 :0 ) have a neutral or cholesterol-decreasing effect . The DNA sequence analysis of the genes ( O75907 , SCAP , P37231 , OLR , P05413 and PRL ) in a random panel of 8 Sirohi goats revealed 38 SNPs across the targeted regions . Out of the studied SNPs ( 38 ) across these genes , 22 SNPs had significant effect on one or a group of fatty acids including DB01211 . The genotypes at these loci showed significant differences in the least square means of a particular fatty acid or a group of fatty acids including DB01211 and its isomers . The unexpected effect of cyclosporin A on CD56+CD16- and CD56+CD16+ natural killer cell subpopulations . DB00091 ( Q13216 ) is commonly used to prevent graft-versus-host disease . The influence of Q13216 on T-cell function has been extensively investigated ; however , the effect of Q13216 on natural killer ( NK ) cells is less understood . NK cells were cultured with P60568 and P40933 with and without Q13216 for 1 week . Compared with controls , Q13216 -treated cultures showed fewer CD56(+)CD16(+) P55040 (+) NK cells and a reciprocal increase in CD56(+)CD16(-) P55040 (-) cells . These changes were due mainly to a reduced proliferation of the CD56(dim) NK-cell subpopulation and a relative resistance of CD56(bright) NK cells to Q13216 . Following coculture with K562 targets , Q13216 -exposed NK cells differed from controls and lacked Ca(2+) oscillations , nuclear factor of activated T cells ( NFAT ) dephosphorylation , and NFAT nuclear translocation . NK cells cultured in Q13216 retained cytotoxicity against K562 , Raji , and P55040 ligand-expressing lymphoblastoid cells . NK cells cultured in Q13216 showed increases in O14931 and reductions in O95944 and P26718 . Following IL-12 and Q14116 stimulation , Q13216 -treated NK cells showed more P01579 -producing cells . Using in vitro NK-cell differentiation , progenitor cells gave rise to more CD56(+) P55040 (-) NK cells in the presence of Q13216 than controls . Collectively , these studies show that Q13216 influences NK-cell function and phenotype , which may have important implications for graft-versus-leukemia effects . c-Fos protein as a target of anti-osteoclastogenic action of vitamin D , and synthesis of new analogs . Although active vitamin D drugs have been used for the treatment of osteoporosis , how the vitamin D receptor ( P11473 ) regulates bone cell function remains largely unknown . Using osteoprotegerin-deficient mice , which exhibit severe osteoporosis due to excessive receptor activator of NF-kappaB ligand/receptor activator of NF-kappaB ( O14788 / Q9Y6Q6 ) stimulation , we show herein that oral treatment of these mice with 1alpha,25-dihydroxyvitamin D3 [ DB00136 ] inhibited bone resorption and prevented bone loss , suggesting that P11473 counters O14788 / Q9Y6Q6 signaling . In P09603 -dependent osteoclast precursor cells isolated from mouse bone marrow , DB00136 potently and dose-dependently inhibited their differentiation into multinucleate osteoclasts induced by O14788 . Among signaling molecules downstream of Q9Y6Q6 , DB00136 inhibited the induction of c-Fos protein after O14788 stimulation , and retroviral expression of c-Fos protein abrogated the suppressive effect of DB00136 on osteoclast development . By screening vitamin D analogs based on their c-Fos-suppressing activity , we identified a new analog , named DD281 , that inhibited bone resorption and prevented bone loss in ovariectomized mice , more potently than DB00136 , with similar levels of calcium absorption . Thus , c-Fos protein is an important target of the skeletal action of P11473 -based drugs , and DD281 is a bone-selective analog that may be useful for the treatment of bone diseases with excessive osteoclastic activity . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . DB00136 induces capacitative calcium entry involving a Q13507 protein in skeletal muscle and osteoblastic cells . This work describes the involvement of TRPC proteins in capacitative calcium entry ( CCE ) induced by 1alpha,25-dihydroxy-vitamin-D3 [ DB00136 ] in chick skeletal muscle and in rat osteoblast-like cells ( ROS 17/2.8 ) and the role of the vitamin D receptor ( P11473 ) in this non-genomic rapid response mediated by the hormone . We propose that an endogenous Q13507 protein mediates DB00136 modulation of CCE in these cells , which seems to implicate P11473 - Q13507 association and the participation of an INAD-like scaffold protein . Stimulation of the peroxisome proliferator-activated receptor gamma ( Q07869 gamma ) and the expression of selected blood monocyte cytokine genes in diabetic macroangiopathy . Monocytes and macrophages play a key role in the progression of atheromatous changes . The peroxisome proliferator-activated receptor gamma ( Q07869 gamma ) can limit macroangiopathy through the control of cytokine transcription . The objectives of this study were to examine the influence of Q07869 gamma and its agonist ( rosiglitazone ) on the TNFalpha , P05231 , P10145 and P22301 gene expression in monocytes of patients with diabetic macroangiopathy and to analyse obtained results in context of selected atherogenic factors ant direct indicators of endothelial lesion . TNFalpha , P05231 , P10145 , P22301 and Q07869 gamma gene expression was assessed in peripheral blood monocytes in 45 patients with type 2 diabetes before and following 22 weeks of rosiglitazone therapy ( real-time PCR [ Applied Biosystems ] ) . As indicators of endothelial lesion , concentration of thrombomodulin ( immunoassay [ Diagnostica Stago ] ) and amount of circulating blood endothelial cells ( immunofluorescence method with MoAb Q8N0X4 -HEC19 ) were determined . Following rosiglitazone therapy , a statistically significant downward tendency of TNFalpha ( p=0.026 ) and P10145 ( p=0.008 ) gene expression was noted . Before and following rosiglitazone treatment , Q07869 gamma , P05231 and P22301 gene expression was undetectable in studied monocytes in vivo . In conclusion , TNFalpha and P10145 play an important role in monocyte atherogenic activity . Rosiglitazone reduces monocyte proinflammatory readiness by influencing the expression of selected atherogenic cytokines ( Q07869 gamma-independent pathway ) . Chronic inhibition of farnesyl pyrophosphate synthase attenuates cardiac hypertrophy and fibrosis in spontaneously hypertensive rats . P14324 ( FPPS ) , an essential enzyme in the mevalonate pathway , was reported to be upregulated in young spontaneously hypertensive rats ( SHR ) when compared with Wistar-Kyoto ( WKY ) rats , and this was accompanied by development of left ventricular hypertrophy . Five-week-old rats were daily gavaged with vehicle or an FPPS inhibitor ( alendronate , 1 or 10 mg/kg ) and blood pressures was monitored by the tail-cuff method every other week . Twelve weeks of alendronate treatment attenuated the left ventricular weight to body weight ratio ( LVW/BW ) , hydroxyproline content , collagen deposition in the interstitia , and gene expression of atrial natriuretic peptide , B-type natriuretic peptide , and procollagen type I/III in the SHR left ventricle , all of which were significantly higher in SHRs than in WKY rats . Furthermore , long-term treatment with an FPPS inhibitor significantly reduced RhoA activation , P29323 phosphorylation , and TGF-beta1 expression in the SHR left ventricle , all of which were upregulated more in SHRs than in WKY rats . In conclusion , chronic treatment with an FPPS inhibitor attenuates the development of cardiac hypertrophy and fibrosis , and the suppression of P27361 /2 phosphorylation and TGF-beta1 expression with inhibition of RhoA activation may be an important mechanism . Characterization of the antitumor activity of a polyantigenic immunomodulator ( P05121 ) : I -- Utilization of cimetidine and cyclosporine A . DB00501 ( CMT ) , cyclosporine A ( DB00091 ) , interleukin 2 ( IL 2 ) , were used to characterize the anticancerous effect of a polyantigenic immunodulator ( P05121 ) . P05121 consists of a mixture of inactivated bacteria and influenza virus in a peanut oil-arlacel A-aluminum monoesterate emulsion . Antitumoral activity was tested on Ehrlich 's ascites tumor ( EAT ) implanted into Swiss-Webster ( allogeneic ) or C57BL/6J ( syngeneic ) mice . P05121 antitumor activity was enhanced by CMT acting synergistically by reducing substantially the tumor growth and increasing the percentage of surviving mice , while DB00091 suppressed this activity . P05121 or its individual components were able to induce significant lymphocyte blastogenesis in mouse ( C57BL/6J ) -spleen lymphocytes and P60568 enhanced considerably this lymphoproliferative response . The results suggest that P05121 acts at the level of cellular immunity . Selective use of multiple vitamin D response elements underlies the 1 alpha,25-dihydroxyvitamin D3-mediated negative regulation of the human O15528 gene . The human 25-hydroxyvitamin D3 ( DB00146 ) 1alpha-hydroxylase , which is encoded by the O15528 gene , catalyzes the metabolic activation of the DB00146 into 1alpha,25-dihydroxyvitamin D3 ( DB00136 ) , the most biologically potent vitamin D3 metabolite . The most important regulator of O15528 gene activity is DB00136 itself , which down-regulates the gene . The down-regulation of the O15528 gene has been proposed to involve a negative vitamin D response element ( nVDRE ) that is located approximately 500 bp upstream from transcription start site ( TSS ) . In this study , we reveal the existence of two new P11473 -binding regions in the distal promoter , 2.6 and 3.2 kb upstream from the TSS , that bind vitamin D receptor-retinoid X receptor complexes . Since the down regulation of the O15528 gene is tissue- and cell-type selective , a comparative study was done for the new DB00136 -responsive regions in P29320 -293 human embryonic kidney and MCF-7 human breast cancer cells that reflect tissues that , respectively , are permissive and non-permissive to the phenomenon of DB00136 -mediated down-regulation of this gene . We found significant differences in the composition of protein complexes associated with these O15528 promoter regions in the different cell lines , some of which reflect the capability of transcriptional repression of the O15528 gene in these different cells . In addition , chromatin architecture differed with respect to chromatin looping in the two cell lines , as the new distal regions were differentially connected with the proximal promoter . This data explains , in part , why the human O15528 gene is repressed in P29320 -293 but not in MCF-7 cells . [ Moclobemide ( DB01171 ) , the first P21397 -inhibitor : really something new ? ] . Role of vitamin D receptor in the antiproliferative effects of calcitriol in tumor-derived endothelial cells and tumor angiogenesis in vivo . Calcitriol ( DB00136 ) , the major active form of vitamin D , is antiproliferative in tumor cells and tumor-derived endothelial cells ( TDEC ) . These actions of calcitriol are mediated at least in part by vitamin D receptor ( P11473 ) , which is expressed in many tissues including endothelial cells . To investigate the role of P11473 in calcitriol effects on tumor vasculature , we established TRAMP-2 tumors subcutaneously into either P11473 wild-type ( WT ) or knockout ( KO ) mice . Within 30 days post-inoculation , tumors in KO mice were larger than those in WT ( P < 0.001 ) . TDEC from WT expressed P11473 and were able to transactivate a reporter gene whereas TDEC from KO mice were not . Treatment with calcitriol resulted in growth inhibition in TDEC expressing P11473 . However , TDEC from KO mice were relatively resistant , suggesting that calcitriol-mediated growth inhibition on TDEC is P11473 -dependent . Further analysis of the TRAMP- P06681 tumor sections revealed that the vessels in KO mice were enlarged and had less pericyte coverage compared with WT ( P < 0.001 ) . Contrast-enhanced magnetic resonance imaging showed an increase in vascular volume of TRAMP tumors grown in P11473 KO mice compared with WT mice ( P < 0.001 ) and FITC-dextran permeability assay suggested a higher extent of vascular leakage in tumors from KO mice . Using ELISA and Western blot analysis , there was an increase of hypoxia-inducible factor-1alpha , vascular endothelial growth factor , angiopoietin 1 , and platelet-derived growth factor-BB levels observed in tumors from KO mice . These results indicate that calcitriol-mediated antiproliferative effects on TDEC are P11473 -dependent and loss of P11473 can lead to abnormal tumor angiogenesis .

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DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . DB00171 -sensitive potassium channels ( K( DB00171 ) ) in retina : a key role for delayed ischemic tolerance . The objectives of the present study were to determine the localization of K( DB00171 ) channels in normal retina and to evaluate their potential roles in ischemic preconditioning ( IPC ) in a rat model of ischemia induced by increased intraocular pressure ( IOP ) . Brown Norway rats were subjected to sublethal 3- , lethal 20- and 40-min ischemia and the functional recovery was evaluated using electroretinography . The time interval between ischemic insults ranged from 1 to 72 h . The effects of K( DB00171 ) channel blockade on IPC protection were studied by treatment with 0.01 % glipizide . IPC was mimicked by injection of K( DB00171 ) channel openers of 0.01 % (-)cromakalim or 0.01 % P1060 72 h before 20-min ischemia . Co-expression of K( DB00171 ) channel subunits Kir6.2/ Q09428 was observed in the retinal pigment epithelium , inner segments of photoreceptors , outer plexiform and ganglion cell layers and at the border of the inner nuclear layer . In contrast to a 20- or 40-min ischemia , a 3-min ischemia induced no alteration of the electroretinogram ( ERG ) and constituted the preconditioning stimulus . An ischemic challenge of 40 min in preconditioned rats induced impairment of retinal function . However , animals preconditioned 24 , 48 and 72 h before 20-min ischemia had a significant improvement of the ERG . (-)Cromakalim and P1060 mimicked the effect of IPC . DB01067 significantly suppressed the protective effects of preconditioning . In conclusion , activation of K( DB00171 ) channels plays an important role in the mechanism of preconditioning by enhancing the resistance of the retina against a severe ischemic insult . The genomic organization of the human P43220 gene . The genomic organization of the human gene encoding the receptor for glucagon-like peptide-1 ( P0C6A0 ( 7-37 ) /(7-36) amide ) was analyzed to reveal the relationship to other G-protein-coupled receptors . The coding sequence of the P43220 is interrupted by 12 introns . These introns are uniformly distributed within the open reading frame . The length of the introns varies between 6.6 kb and 100 bp , in contrast to the relative constant length of 100 bp of the exons . All of the exon/intron splice junctions characterized followed the consensus GT-AG rule . A comparison of the genomic structure with other related receptor genes indicates that the exon/intron organization is well-conserved among the P01282 / glucagon/secretin receptor family . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . Neuroprotective effect of the glucagon-like peptide-1 receptor agonist , synthetic exendin-4 , in streptozotocin-induced diabetic rats . BACKGROUND AND PURPOSE : Glucagon-like peptide-1 ( P0C6A0 ) receptors are widely expressed in neural tissues and diminish neuronal degeneration or induce neuronal differentiation . The aim of this study was to investigate the effect of the P0C6A0 pathway on peripheral nerves in streptozotocin-induced diabetic rats . EXPERIMENTAL APPROACH : Diabetic and nondiabetic rats were treated with the P43220 agonist , synthetic exendin-4 ( i.p. , 1 nmol·kg(-1)·day(-1) ) or placebo for 24 weeks , and current perception threshold values , DB02527 levels and nerve fibre size in the sciatic nerve were measured . We also investigated P43220 expression , quantitative changes in P09936 -positive intraepidermal nerve fibres and cleaved caspase 3-stained Schwann cells by immunohistochemistry . KEY RESULTS : P43220 expression was detected in the sciatic nerve and skin . After exendin-4 treatment , the increase seen in current perception threshold values at 2000 and 250 Hz in diabetic rats was reduced . Also , the decrease in myelinated fibre size or axon/fibre area ratio in the sciatic nerve and the loss of intraepidermal nerve fibre in the skin of diabetic rats were ameliorated . These responses were closely associated with the attenuation of Schwann cell apoptosis and improvement in the DB02527 level in exendin-4-treated diabetic rats , compared with placebo-treated animals . CONCLUSION AND IMPLICATIONS : DB01276 may prevent peripheral nerve degeneration induced by diabetes in an animal model , supporting the hypothesis that P0C6A0 may be useful in peripheral neuropathy . The neuroprotection is probably attributable to P43220 activation , antiapoptotic effects and restoration of DB02527 content . P01308 -like growth factor-1 receptor inhibitor , Q99217 -479 , in cetuximab-refractory head and neck squamous cell carcinoma . BACKGROUND : Recurrent head and neck squamous cell carcinoma ( HNSCC ) remains a difficult cancer to treat . Here , we describe a patient with HNSCC who had complete response to methotrexate ( MTX ) after progressing on multiple cytotoxic agents , cetuximab , and Q99217 -479 ( monoclonal antibody against insulin-like growth factor-1 receptor [ IGF-1R ] ) . METHODS : The clinical information was collected by a retrospective medical record review under an Institutional Review Board-approved protocol . From 4 tumors and 2 normal mucosal epithelia , global gene expression , and IGF-1R and dihydrofolate reductase ( P00374 ) protein levels were determined . RESULTS : Effective target inhibition in the tumor was confirmed by the decreased protein levels of total and phospho-IGF-1R after treatment with Q99217 -479 . Decreased level of P00374 and conversion of a gene expression profile associated with cetuximab-resistance to cetuximab-sensitivity were also observed . CONCLUSION : This suggests that the combination of Q99217 -479 and MTX or cetuximab may be a promising therapeutic approach in refractory HNSCC . HRAS1 and P01308 genes are relocated but not structurally altered as a result of the t(7;11)(p15;p15) in a clone from a patient with acute myeloid leukaemia ( M4 ) . A patient whose leukaemic cells carried the rare t(7;11)(p15;p15) was diagnosed as having acute myelomonocytic leukaemia ( AML-M4 ) , and supports the association of this specific translocation with forms of acute myeloid leukaemia showing differentiation . Blast phase chronic myeloid leukaemia was excluded by lack of involvement of the P00519 and P11274 genes . Chromosome in situ hybridization studies showed that both the HRAS1 and P01308 genes were present on the terminal part of chromosome 11p which was translocated to chromosome 7p . Neither HRAS1 nor P01308 were structurally rearranged . Field inversion gel electrophoresis showed that a 400 kb fragment encompassing HRAS1 was structurally entire in leukaemic DNA . Because the P01308 gene , which was also translocated , is probably located proximal to HRAS1 on chromosome 11p , it is unlikely that HRAS1 was near the chromosome 11 breakpoint or involved in this leukaemia . New insights into the role of DB02527 in the production and function of the incretin hormone glucagon-like peptide-1 ( P0C6A0 ) . The proglucagon gene ( gcg ) encodes both glucagon and glucagon-like peptide-1 ( P0C6A0 ) , produced in pancreatic alpha cells and intestinal endocrine L cells , respectively . The incretin hormone P0C6A0 stimulates insulin secretion and pro-insulin gene transcription . P0C6A0 also enhances pancreatic beta-cell proliferation , inhibits cell apoptosis , and has been utilized in the trans-differentiation of insulin producing cells . A long-term effective P43220 agonist , DB01276 , has now been developed as the drug in treating type II diabetes and potentially other metabolic disorders . The expression of gcg and the production of P0C6A0 can be activated by the elevation of the second messenger cyclic AMP ( DB02527 ) . Recent studies suggest that in addition to protein kinase A ( PKA ) , exchange protein activated by DB02527 ( Epac ) , another effector of DB02527 , and the crosstalk between PKA and the Wnt signaling pathway , are involved in DB02527 -stimulated gcg transcription and P0C6A0 production as well . Finally , functions of P0C6A0 in pancreatic beta cells are also mediated by PKA , Epac , as well as the effector of the Wnt signaling pathway . Together , these novel findings bring us a new insight into the role of DB02527 in the production and function of the incretin hormone P0C6A0 . Signaling and biological effects of glucagon-like peptide 1 on the differentiation of DB05914 from human bone marrow . Glucagon-like peptide 1 ( P0C6A0 ) functions as an incretin hormone with antidiabetogenic properties . However , the role of P0C6A0 in human bone marrow-derived DB05914 ( hMSCs ) , if any , remains unknown . The effects of P0C6A0 on hMSCs were tested with regard to cell proliferation , cytoprotection , and cell differentiation into adipocytes . The signaling pathways involved in these processes were also analyzed . Cells were characterized with biochemical and morphological approaches before and after being induced to differentiate into adipocytes . P12004 protein levels were used as a proliferation index , whereas cell apoptosis was studied by deprivation of fetal bovine serum . Isolated hMSCs expressed stem cell markers as well as mRNA and P43220 protein . P0C6A0 increased the proliferation of hMSCs , which decreased when they were induced to differentiate into adipocytes . This process produced biochemical and morphological changes in cells expressing PPARgamma , C/EBPbeta , AP2 , and P06858 in a time-dependent pattern . Notably , P0C6A0 significantly reduced the expression of PPARgamma , C/EBPbeta , and P06858 . These effects were exerted at least through the MEK and PKC signaling pathways . In addition , P0C6A0 significantly reduced cell apoptosis . Our data indicate that , in hMSCs , P0C6A0 promotes cellular proliferation and cytoprotection and prevents cell differentiation into adipocytes . These latter findings underscore the potential therapeutic role of P0C6A0 in preventing the adipocyte hyperplasia associated with obesity and , additionally , could bolster the maintenance of hMSC stores by promoting the proliferation and cytoprotection of undifferentiated hMSC . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . Enhanced sensitivity to irinotecan by Cdk1 inhibition in the p53-deficient HT29 human colon cancer cell line . Mutations in the tumor-suppressor gene p53 have been associated with advanced colorectal cancer ( CRC ) . DB00762 ( CPT-11 ) , a P11387 inhibitor , has been recently incorporated to the adjuvant therapy . Since the DNA-damage checkpoint depends on p53 activation , the status of p53 might critically influence the response to CPT-11 . We analysed the sensitivity to CPT-11 in the human colon cancer cell line HT29 ( mut p53 ) and its wild-type (wt)-p53 stably transfected subclone HT29-A4 . Cell-cycle analysis in synchronised cells demonstrated the activation of transfected wt-p53 and a P38936 ( P38936 /CIP1)-dependent cell-cycle blockage in the S phase . Activated wt-p53 increased apoptosis and enhanced sensitivity to CPT-11 . In p53-deficient cells , cDNA-macroarray analysis and western blotting showed an accumulation of the cyclin-dependent kinase (cdk)1/cyclin B complex . Subsequent p53-independent activation of the cdk-inhibitor ( cdk-I ) P38936 ( P38936 /CIP1) prevented cell-cycle progression . Cdk1 induction was exploited in vivo to improve the sensitivity to CPT-11 by additional treatment with the cdk-I P99999 -202 . We demonstrate a gain of sensitivity to CPT-11 in a p53-mutated colon cancer model either by restoring wild-type p53 function or by sequential treatment with cdk-Is . Considering that mutations in p53 are among the most common genetic alterations in CRC , a therapeutic approach specifically targeting p53-deficient tumors could greatly improve the treatment outcomes . Molecular physiology of glucagon-like peptide-1 insulin secretagogue action in pancreatic β cells . P01308 secretion from pancreatic β cells is stimulated by glucagon-like peptide-1 ( P0C6A0 ) , a blood glucose-lowering hormone that is released from enteroendocrine L cells of the distal intestine after the ingestion of a meal . P0C6A0 mimetics ( e.g. , DB01276 ) and P0C6A0 analogs ( e.g. , DB06655 ) activate the β cell P43220 ( P43220 ) , and these compounds stimulate insulin secretion while also lowering levels of blood glucose in patients diagnosed with type 2 diabetes mellitus ( T2DM ) . An additional option for the treatment of T2DM involves the administration of dipeptidyl peptidase-IV ( DPP-IV ) inhibitors ( e.g. , Januvia , DB04876 ) . These compounds slow metabolic degradation of intestinally released P0C6A0 , thereby raising post-prandial levels of circulating P0C6A0 substantially . Investigational compounds that stimulate P0C6A0 secretion also exist , and in this regard a noteworthy advance is the demonstration that small molecule Q8TDV5 agonists ( e.g. , AR231453 ) stimulate L cell P0C6A0 secretion while also directly stimulating β cell insulin release . In this review , we summarize what is currently known concerning the signal transduction properties of the β cell P43220 as they relate to insulin secretion . Emphasized are the cyclic AMP , protein kinase A , and Epac2-mediated actions of P0C6A0 to regulate DB00171 -sensitive K⁺ channels , voltage-dependent K⁺ channels , O94759 cation channels , intracellular Ca⁺ release channels , and Ca⁺-dependent exocytosis . We also discuss new evidence that provides a conceptual framework with which to understand why P43220 agonists are less likely to induce hypoglycemia when they are administered for the treatment of T2DM . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Novel P0C6A0 mimetics developed to treat type 2 diabetes promote progenitor cell proliferation in the brain . One of the symptoms of diabetes is the progressive development of neuropathies . One mechanism to replace neurons in the CNS is through the activation of stem cells and neuronal progenitor cells . We have tested the effects of the novel P0C6A0 mimetics exenatide ( exendin-4 ; DB01276 ) and liraglutide ( DB06655 ; DB06655 ) , which are already on the market as treatments for type 2 diabetes , on the proliferation rate of progenitor cells and differentiation into neurons in the dentate gyrus of brains of mouse models of diabetes . P0C6A0 analogues were injected subcutaneously for 4 , 6 , or 10 weeks once daily in three mouse models of diabetes : ob/ob mice , db/db mice , or high-fat-diet-fed mice . Twenty-four hours before perfusion , animals were injected with 5'-bromo-2'-deoxyuridine ( BrdU ) to mark dividing progenitor cells . By using immunohistochemistry and stereological methods , the number of progenitor cells or doublecortin-positive young neurons in the dentate gyrus was estimated . We found that , in all three mouse models , progenitor cell division was enhanced compared with nondiabetic controls after chronic i.p. injection of either liraglutide or exendin-4 by 100-150 % ( P < 0.001 ) . We also found an increase in young neurons in the DG of high-fat-diet-fed mice after drug treatment ( P < 0.001 ) . The P43220 antagonist exendin(9-36) reduced progenitor cell proliferation in these mice . The results demonstrate that P0C6A0 mimetics show promise as a treatment for neurodegenerative diseases such as Alzheimer 's disease , because these novel drugs cross the blood-brain barrier and increase neuroneogenesis . A Short-activating RNA Oligonucleotide Targeting the Islet β-cell Transcriptional Factor MafA in P28906 (+) Cells . Upon functional loss of insulin producing islet β-cells , some patients with diabetes become dependent on life-long insulin supplementation therapy . Bioengineering surrogate insulin producing cells is an alternative replacement strategy . We have developed a novel approach using short-activating RNA oligonucleotides to differentiate adult human P28906 (+) cells into insulin-secreting cells . By transfecting RNA to increase transcript levels of the master regulator of insulin biosynthesis , v-maf musculoaponeurotic fibrosarcoma oncogene homolog A ( MafA ) , several pancreatic endodermal genes were upregulated during the differentiation procedure . These included Pancreatic and duodenal homeobox gene-1 ( PDX1 ) , Neurogenin 3 , Q13562 , and NK6 homeobox 1 ( NKx6-1 ) . Differentiated P28906 (+) cells also expressed glucokinase , glucagon-like peptide 1 receptor ( P43220 ) , sulfonylurea receptor-1 ( Q09428 ) and phogrin-all essential for glucose sensitivity and insulin secretion . The differentiated cells appropriately processed C-peptide and insulin in response to increasing glucose stimulation as shown by enzyme-linked immunosorbent assay ( ELISA ) , fluorescence-activated cell sorting analysis , western blotting , and immunofluorescence staining . We provide a new approach using short-activating RNA in developing insulin producing surrogate cells for treating diabetes.Molecular Therapy - Nucleic Acids ( 2013 ) 2 , e97 ; doi:10.1038/mtna.2013.23 ; advance online publication 4 June 2013 . Comparison of two polymer-based immunohistochemical detection systems : ENVISION+ and ImmPRESS . The non-specific background reaction produced in avidin-biotin-based immunohistochemistry , particularly after harsh antigen retrieval procedures , has promoted the use of non-avidin-biotin systems , yet there are few reports comparing the performance of non-avidin-biotin , polymer-based methods . In this study we compare two of these methods , ENVISION+trade mark and ImmPRESS , in animal tissues . We examined the immunoreactivity of 18 antigens in formalin-fixed , paraffin-embedded tissues . Antigens were located in the cytoplasmic membrane ( CD11d , P05107 and CD79a ) , cytoplasm ( calretinin , P23219 , P35354 , Glut-1 , HepPar 1 , P10721 , Melan A , tryptase and uroplakin III ) or nucleus ( Q2TAK8 , P09936 and thyroid transcription factor 1 ) . We also evaluated three infectious agents ( Aspergillus , calicivirus and West Nile virus ) . The staining with ENVISION+ or ImmPRESS was performed simultaneously for each antigen . The intensity of the reaction and background staining were scored . ImmPRESS yielded similar or higher reaction intensity than ENVISION+trade mark in 16/18 antigens . ImmPRESS produced abundant background with the other two antigens ( calretinin and P35354 ) , which hindered interpretation of the specific reaction . The cost of ImmPRESS was 25 % lower than for ENVISION+trade mark . Based on these results , ImmPRESS is a good polymer-based detection system for routine immunohistochemistry . Concurrent pharmacological modification of cannabinoid-1 and glucagon-like peptide-1 receptor activity affects feeding behavior and body weight in rats fed a free-choice , high-carbohydrate diet . To extend preliminary studies on the effects on food intake of the combined use of cannabinoid ( CB ) 1 and glucagon-like peptide-1 ( P0C6A0 ) receptor agonists and antagonists , the effect of these drugs on the feeding behavior in rats maintained on a free-choice , high-carbohydrate diet was investigated over a longer period of time . Rats were fed a standard diet for 3 days and then fed with both the standard and the high-sucrose chow . After 4 days of the high-calorie diet , the following combination treatments were administered daily by an intraperitoneal injection for the next 3 days : 1 mg/kg AM 251 ( a P21554 receptor antagonist ) or 1 mg/kg Q08050 55,212-2 ( a P21554 receptor agonist ) together with 3 µg/kg exendin-4 ( Ex-4 , a P43220 agonist ) or 160 µg/kg exendin ( 9-39 ) [ Ex ( 9-39 ) , a P43220 antagonist ] . The total daily caloric intake and body weight were significantly reduced in rats treated with Ex-4 and AM 251 or Q08050 55,212-2 compared with either of the drugs injected alone and the saline-injected controls . Both drug combinations selectively inhibited ingestion of the high-sucrose chow . Although Ex ( 9-39 ) administration did not significantly affect food consumption , it resulted in a marked body weight gain , indicating that the P43220 antagonist caused a positive energy balance . It is concluded that AM 251 or Q08050 55,212-2 and Ex-4 , injected together , exert additive , inhibitory effects on the consumption of high-sugar food . Exendin-4 alleviates angiotensin II-induced senescence in vascular smooth muscle cells by inhibiting Rac1 activation via a DB02527 /PKA-dependent pathway . Vascular aging has been implicated in the progression of diabetes and age-related cardiovascular disorders . Glucagon-like peptide-1 ( P0C6A0 ) is an incretin hormone capable of cytoprotective actions in addition to its glucose-lowering effect . The present study was undertaken to examine whether Exendin-4 , a specific ligand for the P43220 , could prevent angiotensin ( P03950 ) II-induced premature senescence in vascular smooth muscle cells ( VSMCs ) and to determine the underlying mechanism involved . Senescence-associated β-galactosidase ( SA β-gal ) assay showed that P03950 II induced premature senescence of VSMCs . Pretreatment with Exendin-4 significantly attenuated P03950 II-induced generation of H2O2 and the subsequent VSMC senescence . These effects were , however , reversed in the presence of exendin fragment 9-39 , a P43220 antagonist , or PKI14-22 . Moreover , a marked increase in the levels of p53 and P38936 induced by P03950 II was blunted by the treatment with Exendin-4 . Nevertheless , Exendin-4 failed to decrease P03950 II-induced expression of NAD(P)H oxidase 1 ( Nox1 ) , NAD(P)H oxidase 4 ( Nox4 ) , O75935 (phox) , or p47(phox) in VSMCs . Mechanistically , Exendin-4 blocked P03950 II-induced Rac1 activation through the DB02527 /PKA signaling cascade . Specifically , NSC23766 , a Rac1 inhibitor , abrogated the suppressive effects of Exendin-4 on P03950 II-induced premature senescence and H2O2 generation , respectively . Thus Exendin-4 confers resistance to P03950 II-induced superoxide anion generation from NAD(P)H oxidase and the resultant VSMC senescence by inhibiting Rac1 activation via a DB02527 /PKA-dependent pathway . These findings demonstrate that P0C6A0 as well as its analogs ( P0C6A0 -related reagents ) may hold therapeutic potential in the treatment of diabetes with cardiovascular disease . Chemical coding of the human gastrointestinal nervous system : cholinergic , VIPergic , and catecholaminergic phenotypes . The aim of this investigation was to identify the proportional neurochemical codes of enteric neurons and to determine the specific terminal fields of chemically defined nerve fibers in all parts of the human gastrointestinal ( GI ) tract . For this purpose , antibodies against the vesicular monoamine transporters ( P54219 /2 ) , the vesicular acetylcholine transporter ( Q16572 ) , tyrosine hydroxylase ( TH ) , dopamine beta-hydroxylase ( P09172 ) , serotonin ( 5-HT ) , vasoactive intestinal peptide ( P01282 ) , and protein gene product 9.5 ( P09936 ) were used . For in situ hybridization (35)S-labeled P54219 , Q05940 , and Q16572 riboprobes were used . In all regions of the human GI tract , 50-70 % of the neurons were cholinergic , as judged by staining for Q16572 . The human gut unlike the rodent gut exhibits a cholinergic innervation , which is characterized by an extensive overlap with VIPergic innervation . Neurons containing Q05940 constituted 14-20 % of all intrinsic neurons in the upper GI tract , and there was an equal number of TH-positive neurons . In contrast , P09172 was absent from intrinsic neurons . Cholinergic and monoaminergic phenotypes proved to be completely distinct phenotypes . In conclusion , the chemical coding of human enteric neurons reveals some similarities with that of other mammalian species , but also significant differences . P01282 is a cholinergic cotransmitter in the intrinsic innervation of the human gut . The substantial overlap between Q05940 and TH in enteric neurons indicates that the intrinsic catecholaminergic innervation is a stable component of the human GI tract throughout life . The absence of P09172 from intrinsic catecholaminergic neurons indicates that these neurons have a dopaminergic phenotype . [ DB00988 -beta-hydroxylaseaktivität im Plasma von Dialysepatienten ( author 's transl ) ] . Plasma dopamin-b-hydroxylase ( P09172 ) was studied in 70 healthy control persons and in 37 hemodialysed patients . Basal P09172 in controls corresponded to 50.0 +/- 29.3 IU . There was was no significant difference between males ( 53.9 +/1 33.8 IU ) and females ( 47.4 +/- 25 IU ) ; no correlation could be found between age and plasma P09172 . In hemodialysed patients basal P09172 levels were significantly ( p less than 0.01 ) decreased ( 32.5 % /- 17.6 IU ) , suggesting lowered sympathetic activity and/or abnormalities in release , distribution space , or metabolism of P09172 . During hemodialysis plasma P09172 activity rose during ultrafiltration . This finding indicates a directionally appropriate sympathetic reflex response to volume depletion in dialysed patients . Improvement of psoriasis during exenatide treatment in a patient with diabetes . CONTEXT AND AIM : Psoriasis is an immune-mediated skin disorder frequently associated with obesity and type 2 diabetes ( T2D ) . This report is of a clinically significant improvement in psoriasis lesions in a patient with T2D during treatment with a P43220 agonist ( exenatide ) . OBSERVATION : A 61-year-old male patient ( BMI : 25.5 kg/m(2) ) with T2D treated with metformin and sulphonylureas had also complained , since 1980 , of extensive psoriasis that required multiple steroid-based treatments [ Psoriasis Area and Sensitivity Index ( PASI ) score : 11 ] . In September 2008 , his diabetes treatment was intensified with exenatide ( DB01276 (®) ) to improve poor glycaemic control . The patient , as expected , lost weight and reduced HbA(1c) levels from 65 mmol/mol to 56 mmol/mol . However , after just 1 month of treatment with exenatide , the patient also reported a dramatic improvement in psoriatic plaques that was confirmed at the 1-year follow-up ( PASI : estimated at 3-4 ) . Withdrawal of exenatide was associated with weight gain , deterioration of glycaemic control and deterioration of psoriasis ( PASI: > 10 ) . After reinstating exenatide treatment , the patient again reported a prompt improvement in psoriasis ( PASI : 3.1 ) . CONCLUSION : There was a major and rapid improvement in psoriasis in our patient with T2D following treatment with exenatide . A possible mechanism might be through direct modulation of the immune system by P43220 agonists . 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 . Exenatide twice daily : a review of its use in the management of patients with type 2 diabetes mellitus . Exenatide , administered subcutaneously twice daily ( DB01276 (®) ) , is a synthetic version of the natural peptide exendin-4 , which is a glucagon-like peptide-1 ( P0C6A0 ) receptor agonist ( incretin mimetic ) . Exenatide binds to the P43220 with the same affinity as P0C6A0 , but has a much longer half-life , since it is not degraded by the enzyme dipeptidyl peptidase-4 . Exenatide twice daily enhances glucose-dependent insulin secretion , suppresses inappropriately elevated glucagon secretion , slows gastric emptying and reduces caloric intake . In well-designed clinical trials , adjunctive subcutaneous exenatide 5 or 10 μg twice daily for 16-52 weeks significantly and dose-dependently improved glycaemic control and reduced mean body weight compared with placebo in patients with type 2 diabetes inadequately controlled with oral antihyperglycaemic drugs ( OADs ) and/or basal insulin . The improvements in glycaemic control and reductions in body weight were stably maintained during long-term therapy ( up to 3.5 years ) . The efficacy of adjunctive exenatide twice daily was generally similar to that of basal , prandial or biphasic insulin , sulfonylureas , rosiglitazone and lixisenatide , and less than that of liraglutide , taspoglutide or exenatide once weekly with respect to reductions in glycated haemoglobin . Exenatide twice daily was generally well tolerated ; mild to moderate nausea and vomiting , which decreased with time on therapy , were the most common adverse events . In patients not receiving concomitant sulfonylureas or insulin , the incidence of hypoglycaemia was low ; when it did occur , it was generally mild in severity . Thus , adjunctive exenatide twice daily is a valuable option in the treatment of type 2 diabetes inadequately controlled with OADs and/or basal insulin . [ DB01276 : first once weekly P43220 agonist ( exenatide P10586 ) ] . DB01276 is a new galenic formulation ( long-acting release ) of exenatide , the first agonist of Glucagon-Like Peptide-1 ( P0C6A0 ) receptors having been commercialized for the management of type 2 diabetes . The microsphere technology permits a prolonged absorption of exenatide from the subcutaneous depot , which allows one injection per week instead of two injections per day with the initial formulation of exenatide ( DB01276 ) . The clinical development programme DURATION showed that exenatide 2 mg once weekly more markedly reduces glycated haemoglobin ( HbA(1c) ) , with a similar weight loss but a better digestive tolerance profile ( less nausea and vomiting after treatment initiation ) , compared with the twice daily 10 microg exenatide . When compared to other glucose-lowering agents , once weekly exenatide is more efficacious than sitagliptin , pioglitazone or basal insulin ( glargine or detemir ) , with the advantage of producing weight loss and lowering arterial blood pressure . It does not induce hypoglycaemia and does not necessarily require home blood glucose monitoring , two advantages compared with insulin therapy . DB01276 is currently only reimbursed in Belgium after failure of and in addition to metformin-sulfonylurea combination . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction .

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interacts_with DB01064?

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P25021 activation exacerbates myocardial ischemia/reperfusion injury by disturbing mitochondrial and endothelial function . There is evidence that P25021 blockade improves ischemia/reperfusion ( I/R ) injury , but the underlying cellular mechanisms remain unclear . DB11320 is known to increase vascular permeability and induce apoptosis , and these effects are closely associated with endothelial and mitochondrial dysfunction , respectively . Here , we investigated whether activation of the histamine H2 receptor ( P25021 ) exacerbates myocardial I/R injury by increasing mitochondrial and endothelial permeability . Serum histamine levels were measured in patients with coronary heart disease , while the influence of P25021 activation was assessed on mitochondrial and endothelial function in cultured cardiomyocytes or vascular endothelial cells , and myocardial I/R injury in mice . The serum histamine level was more than twofold higher in patients with acute myocardial infarction than in patients with angina or healthy controls . In neonatal rat cardiomyocytes , histamine dose-dependently reduced viability and induced apoptosis . Mitochondrial permeability and the levels of p- P27361 /2 , Bax , p- Q9UIK4 , and caspase 3 were increased by P25021 agonists . In cultured human umbilical vein endothelial cells ( HUVECs ) , P25021 activation increased p- P27361 /2 and p-moesin levels and also enhanced permeability of HUVEC monolayer . All of these effects were abolished by the P25021 blocker famotidine or the P29323 inhibitor U0126 . After I/R injury or permanent ischemia , the infarct size was reduced by famotidine and increased by an P25021 agonist in wild-type mice . In P25021 KO mice , the infarct size was smaller ; myocardial p- P27361 /2 , p- Q9UIK4 , and mitochondrial Bax were downregulated . These findings indicate that P25021 activation exaggerates myocardial I/R injury by promoting myocardial mitochondrial dysfunction and by increasing cardiac vascular endothelial permeability . Induction of high mobility group box 1 release from serotonin-stimulated human umbilical vein endothelial cells . High mobility group box 1 ( P09429 ) is a non-histone nuclear protein which is released from the nucleus of activated macrophages into the extracellular space in response to stimuli such as endotoxin or necrosis . The P09429 functions as a potent proinflammatory cytokine in the extracellular spaces . Recently , P09429 has been implicated in the progression of atherosclerosis . However , the association between P09429 and the development of atherosclerosis is poorly understood . Therefore , we examined whether serotonin ( 5-HT ) , a key factor involved in the development of atherosclerosis , induced P09429 release in human umbilical vein endothelial cells ( HUVECs ) . We found that 5-HT induced the release of P09429 but not of P27361 /2 and JNK from HUVECs via the 5-HT receptor ( P28222 ) /p38 mitogen-activated protein kinase ( MAPK ) signaling pathway . The p38MAPK inhibitor SB203580 and the P28222 antagonist GR55526 markedly inhibited P09429 release from 5-HT-stimulated HUVECs . The vascular endothelial growth factor ( P15692 ) derived from activated macrophages in atherosclerotic lesions also plays an important role in the progression of atherosclerosis . We found that P09429 induced P15692 production in macrophage-like RAW264.7 cells . P09429 induced the activation of p38MAPK , P27361 /2 and Akt . The P19957 -kinase inhibitor LY294002 significantly inhibited P15692 production in P09429 -stimulated macrophages , while other kinase inhibitors did not . These results suggest that P09429 release may contribute as a risk factor in the development and progression of atherosclerosis . P11362 - 5-hydroxytryptamine 1A heteroreceptor complexes and their enhancement of hippocampal plasticity . BACKGROUND : The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity . METHODS : The analysis was made with bioluminescence resonance energy transfer , co-immunoprecipitation , in situ proximity ligation assay , binding assay , in cell western and the forced swim test . RESULTS : Using bioluminescence resonance energy transfer analysis , fibroblast growth factor receptor 1 ( P11362 ) -5-hydroxytryptamine 1A ( P08908 ) receptor complexes have been demonstrated and their specificity and agonist modulation characterized . Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location . In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 ( P09038 ) and a P08908 agonist , and dependent on the heteroreceptor interface . In vitro and in vivo studies also revealed a P08908 agonist induced phosphorylation of P11362 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing P09038 levels . Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface . The combined acute and repeated intracerebroventricular treatment with P09038 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test . CONCLUSIONS : The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may , in part , be mediated by activation of the P08908 receptor protomer in the hippocampal P11362 - P08908 receptor complex enhancing the P11362 signaling . Identification of an acetylation-dependant P12956 /FLIP complex that regulates FLIP expression and HDAC inhibitor-induced apoptosis . FLIP is a potential anti-cancer therapeutic target that inhibits apoptosis by blocking caspase 8 activation by death receptors . We report a novel interaction between FLIP and the DNA repair protein P12956 that regulates FLIP protein stability by inhibiting its polyubiquitination . Furthermore , we found that the histone deacetylase ( HDAC ) inhibitor DB02546 ( DB02546 ) enhances the acetylation of P12956 , thereby disrupting the FLIP/ P12956 complex and triggering FLIP polyubiquitination and degradation by the proteasome . Using in vitro and in vivo colorectal cancer models , we further demonstrated that DB02546 -induced apoptosis is dependant on FLIP downregulation and caspase 8 activation . In addition , an Q9UBN7 -specific inhibitor Tubacin recapitulated the effects of DB02546 , suggesting that Q9UBN7 is a key regulator of P12956 acetylation and FLIP protein stability . Thus , HDAC inhibitors with anti- Q9UBN7 activity act as efficient post-transcriptional suppressors of FLIP expression and may , therefore , effectively act as ' FLIP inhibitors ' . Selective cyclooxygenase-2 blocker delays healing of esophageal ulcers in rats and inhibits ulceration-triggered c- DB00134 /hepatocyte growth factor receptor induction and extracellular signal-regulated kinase 2 activation . Nonsteroidal anti-inflammatory drugs , both nonselective and cyclooxygenase-2 ( P35354 ) selective , delay gastric ulcer healing . Whether they affect esophageal ulcer healing remains unexplored . We studied the effects of the P35354 selective inhibitor , celecoxib , on esophageal ulcer healing as well as on the cellular and molecular events involved in the healing process . Esophageal ulcers were induced in rats by focal application of acetic acid . Rats with esophageal ulcers were treated intragastrically with either celecoxib ( 10 mg/kg , once daily ) or vehicle for 2 or 4 days . Esophageal ulceration triggered increases in : esophageal epithelial cell proliferation ; expression of P35354 ( but not P23219 ) ; hepatocyte growth factor ( P14210 ) and its receptor , c- DB00134 ; and activation of extracellular signal-regulated kinase 2 ( P28482 ) . Treatment with celecoxib significantly delayed esophageal ulcer healing and suppressed ulceration-triggered increases in esophageal epithelial cell proliferation , c- DB00134 mRNA and protein expression , and P28482 activity . In an ex vivo organ-culture system , exogenous P14210 significantly increased P28482 phosphorylation levels in esophageal mucosa . A structural analog of celecoxib , SC-236 , completely prevented this effect . These findings indicate that celecoxib delays esophageal ulcer healing by reducing ulceration-induced esophageal epithelial cell proliferation . These actions are associated with , and likely mediated by , down-regulation of the P14210 /c- DB00134 - P28482 signaling pathway . No significant association between genetic variants in 7 candidate genes and response to methylphenidate treatment in adult patients with ADHD . Results from pharmacogenetic investigations of methylphenidate ( DB00422 ) response in patients with ADHD are still inconsistent , especially among adults . This study investigates the role of genetic variants ( P31645 , P28222 , Q8IWU9 , P09172 , P21917 , P21964 , and P60880 ) in the response to DB00422 in a sample of 164 adults . Genes were chosen owing to previous evidence for an influence in ADHD susceptibility . No significant differences in allele or genotype frequencies between DB00422 responders and nonresponders were detected . In conclusion , our findings do not support an effect of these genes in the pharmacogenetics of DB00422 among adults with ADHD . Phosphorylation of beta-arrestin2 regulates its function in internalization of beta(2)-adrenergic receptors . Beta-arrestins mediate agonist-dependent desensitization and internalization of G protein-coupled receptors . Previously , we have shown that phosphorylation of beta-arrestin1 by ERKs at DB00133 -412 regulates its association with clathrin and its function in promoting clathrin-mediated internalization of the receptor . In this paper we report that beta-arrestin2 is also phosphorylated , predominantly at residues DB00156 -383 and DB00133 -361 . DB01064 stimulation of the beta(2)-adrenergic receptor promotes dephosphorylation of beta-arrestin2 . Mutation of beta-arrestin2 phosphorylation sites to aspartic acid decreases the association of beta-arrestin2 with clathrin , thereby reducing its ability to promote internalization of the beta(2)-adrenergic receptor . Its ability to bind and desensitize the beta(2)-adrenergic receptor is , however , unaltered . These results suggest that , analogous to beta-arrestin1 , phosphorylation/dephosphorylation of beta-arrestin2 regulates clathrin-mediated internalization of the beta(2)-adrenergic receptor . In contrast to beta-arrestin1 , which is phosphorylated by P27361 and P28482 , phosphorylation of beta-arrestin2 at DB00156 -383 is shown to be mediated by casein kinase II . Recently , it has been reported that phosphorylation of visual arrestin at DB00133 -366 prevents its binding to clathrin . Thus it appears that the function of all arrestin family members in mediating internalization of G protein-coupled receptors is regulated by distinct phosphorylation/dephosphorylation mechanisms . Regulation of extracellular signal-regulated kinase 1/2 influences hippocampal neuronal survival in a rat model of diabetic cerebral ischemia . Activation of extracellular signal-regulated kinase 1/2 has been demonstrated in acute brain ischemia . We hypothesized that activated extracellular signal-regulated kinase 1/2 can protect hippocampal neurons from injury in a diabetic model after cerebral ischemia/reperfusion . In this study , transient whole-brain ischemia was induced by four-vessel occlusion in normal and diabetic rats , and extracellular signal-regulated kinase 1/2 inhibitor ( U0126 ) was administered into diabetic rats 30 minutes before ischemia as a pretreatment . Results showed that the number of surviving neurons in the hippocampal P00915 region was reduced , extracellular signal-regulated kinase 1/2 phosphorylation and P12956 activity were decreased , and pro-apoptotic Bax expression was upregulated after intervention using U0126 . These findings demonstrate that inhibition of extracellular signal-regulated kinase 1/2 activity aggravated neuronal loss in the hippocampus in a diabetic rat after cerebral ischemia/reperfusion , further decreased DNA repairing ability and accelerated apoptosis in hippocampal neurons . P27361 /2 activation plays a neuroprotective role in hippocampal neurons in a diabetic rat after cerebral ischemia/reperfusion . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . DB01064 inhibits fibroblast growth factor-2-induced growth of renal epithelial cells . The signal transduction pathways modulating P09038 effects in renal tubular epithelial cells ( RTEc ) are not completely understood . Since the DB02527 and the mitogen-activated protein kinase ( MAPK ) pathways can modulate the growth of RTEc , we studied whether two DB02527 elevating agents , isoproterenol and 8-bromo- DB02527 , would modulate basic fibroblast growth factor ( P09038 ) induction of MAPK activity ( P28482 ) and cell proliferation in human renal proximal tubular epithelial cells ( RPTEc ) and Madin-Darby canine kidney cells ( MDCK clone EI1 ) . DB01064 , but not P09038 , stimulated DB02527 production in RPTEc and MDCKEI1 cells . P09038 , isoproterenol , and 8-bromo- DB02527 alone increased P28482 activity in both cell types . However , isoproterenol and 8-bromo- DB02527 partially inhibited the P09038 induction of P28482 activity , but only isoproterenol inhibited the proliferation of both cell types . PD098059 ( 25 microM ) , an inhibitor of MAPK kinase ( Q02750 /2 ) , blocked the P09038 mitogenic effects , but did not affect the 8-bromo- DB02527 -induced mitogenic effects in MDCKEI1 cells . These findings suggest that activation of P28482 is required but not sufficient for mitogenesis in RTEc . We conclude that isoproterenol inhibits the growth-promoting effects of P09038 in RTEc via MEK-dependent and -independent pathways . Role of histamine receptors in the effects of histamine on the production of reactive oxygen species by whole blood phagocytes . AIMS : The diverse physiological functions of histamine are mediated through distinct histamine receptors . In this study we investigated the role of P25021 and Q9H3N8 in the effects of histamine on the production of reactive oxygen species by phagocytes in whole blood . MAIN METHODS : Changes in reactive oxygen species ( ROS ) production by whole blood phagocytes after treatment with histamine , Q9H3N8 agonists ( 4-methylhistamine , VUF8430 ) , P25021 agonist ( dimaprit ) and their combinations with Q9H3N8 antagonist ( JNJ10191584 ) and P25021 antagonist ( ranitidine ) were determined using the chemiluminescence ( CL ) assay . To exclude the direct scavenging effects of the studied compounds on the CL response , the antioxidant properties of all compounds were measured using several methods ( TRAP , ORAC , and luminol-HRP-H2O2 based CL ) . KEY FINDINGS : DB11320 , 4-methylhistamine , VUF8430 and dimaprit inhibited the spontaneous and OZP-activated whole blood CL in a dose-dependent manner . On the other hand , only VUF8430 was able to inhibit PMA-activated whole blood CL . DB00863 , but not JNJ10191584 , completely reduced the effects of histamine , 4-methylhistamine and dimaprit . The direct scavenging ability of tested compounds was negligible . SIGNIFICANCE : Our results demonstrate that the inhibitory effects of histamine on ROS production in whole blood phagocytes were caused by P25021 . Our results also suggest that Q9H3N8 agonists in concentrations higher than 10(-6)M may also influence ROS production via binding to P25021 . Differential functional activity of 5-hydroxytryptamine receptor ligands and beta adrenergic receptor antagonists at 5-hydroxytryptamine1B receptor sites in Chinese hamster lung fibroblasts and opossum renal epithelial cells . Functional activity of 5-hydroxytryptamine ( 5-HT ) receptor ligands and beta adrenergic receptor antagonists was studied at P28222 receptor sites in Chinese hamster lung ( CHL ) fibroblasts by measuring two cellular responses : inhibition of forskolin-stimulated cyclic AMP formation and potentiation of basic fibroblast growth ( BFGF ) induced mitogenesis . A good correlation was found between the potency of agonists to inhibit forskolin-induced cyclic AMP formation and their potency to potentiate P09038 -induced thymidine incorporation in CHL fibroblasts . Potent agonist activity was measured with 5-methoxy-3,1,2,3,6-tetrahydro-4-pyidinyl- 1H-indole ( RU 24,969 ) , 5-carboxamidotryptamine ( 5-CT ) , 3-(1,2,5,6)-tetrahydro-4-pyridyl-5-pyrrolo(3,2-b)pyril-5-one ( CP 93,129 ) and 5-HT , whereas sumatriptan displayed weak agonist activity at concentrations different from its binding affinity for P28222 binding sites . In contrast to the observed P28222 receptor-mediated agonist activity in opossum kidney cells for metergoline and the beta adrenergic receptor antagonists : cyanopindolol , 4-(3-tert-butyl-amino-2-hydroxypropoxy)-indole-2 carbonic acid isopropyl ester ( SDZ 21,009 ) , isamoltane , (-)-propranolol and (-)-pindolol , antagonist activity at P28222 receptor sites was yielded in CHL fibroblasts in accordance with the reported observations at rat brain P28222 receptors . Methiothepin was the only compound that antagonized both the opossum kidney cell and CHL fibroblast P28222 receptor-mediated responses although the antagonist effect was more pronounced in CHL fibroblasts. ( ABSTRACT TRUNCATED AT 250 WORDS ) Inhibition of P29323 activation enhances the repair of double-stranded breaks via non-homologous end joining by increasing P78527 activation . Non-homologous end joining ( NHEJ ) is one of the major pathways that repairs double-stranded DNA breaks ( DSBs ) . Activation of DNA-PK is required for NHEJ . However , the mechanism leading to P78527 activation remains incompletely understood . We provide evidence here that the MEK- P29323 pathway plays a role in P78527 -mediated NHEJ . In comparison to the vehicle control ( DB01093 ) , etoposide ( ETOP ) -induced DSBs in MCF7 cells were more rapidly repaired in the presence of U0126 , a specific MEK inhibitor , based on the reduction of γ P16104 and tail moments . Additionally , U0126 increased reactivation of luciferase activity , which resulted from the repair of restriction enzyme-cleaved DSBs . Furthermore , while inhibition of P29323 activation using the dominant-negative MEK1K97M accelerated the repair of DSBs , enforcing P29323 activation with the constitutively active MEK1Q56P reduced DSB repair . In line with MEK activating P27361 and P28482 kinases , knockdown of either P27361 or P28482 increased DSB repair . Consistent with the activation of P78527 being required for NHEJ , we demonstrated that inhibition of P29323 activation using U0126 , MEK1K97M , and knockdown of P27361 or P28482 enhanced ETOP-induced activation of P78527 . Conversely , enforcing P29323 activation by MEK1Q56P reduced ETOP-initiated P78527 activation . Taken together , we demonstrate that P29323 reduces NHEJ-mediated repair of DSBs via attenuation of P78527 activation . Class I histone deacetylase-mediated repression of the proximal promoter of the activity-regulated cytoskeleton-associated protein gene regulates its response to brain-derived neurotrophic factor . We examined the transcriptional regulation of the activity-regulated cytoskeleton-associated protein gene ( Arc ) , focusing on P23560 -induced Arc expression in cultured rat cortical cells . Although the synaptic activity-responsive element ( SARE ) , located -7 kbp upstream of the Arc transcription start site , responded to DB01221 , P23560 , or P09038 , the proximal region of the promoter ( Arc/-1679 ) was activated by P23560 or P09038 , but not by DB01221 , suggesting the presence of at least two distinct Arc promoter regions , distal and proximal , that respond to extracellular stimuli . Specificity protein 4 ( Q02446 ) and early growth response 1 ( P18146 ) controlled Arc/-1679 transcriptional activity via the region encompassing -169 to -37 of the Arc promoter . We found that trichostatin A ( P32119 ) , a histone deacetylase ( HDAC ) inhibitor , significantly enhanced the inductive effects of P23560 or P09038 , but not those of DB01221 on Arc expression . Inhibitors of class I/IIb HDACs , DB02546 , and class I HDACs , MS-275 , but not of class II HDACs , MC1568 , enhanced P23560 -induced Arc expression . The enhancing effect of P32119 was mediated by the region from -1027 to -1000 bp , to which serum response factor ( P11831 ) and Q13547 bound . The binding of Q13547 to this region was reduced by P32119 . Thus , Arc expression was suppressed by class I HDAC-mediated mechanisms via chromatin modification of the proximal promoter whereas the inhibition of HDAC allowed Arc expression to be markedly enhanced in response to P23560 or P09038 . These results contribute to our understanding of the physiological role of Arc expression in neuronal functions such as memory consolidation . Extracellular signal-regulated kinase and the small GTP-binding protein , Rac , contribute to the effects of transforming growth factor-beta1 on gene expression . The kinases and regulatory proteins that convey signals initiated by transforming growth factor-beta ( TGF-beta ) to the nucleus are poorly characterized . To study the role of the extracellular signal-regulated kinase ( P29323 ) pathway in this process , we transiently transfected NIH 3T3 fibroblasts with TGF-beta-responsive luciferase reporter genes and expression vectors designed to interrupt this kinase cascade . Mitogen-activated protein ( Q96HU1 ) kinase phosphatase-1 and a dominant negative Q96HU1 / P29323 kinase 1 mutant reduced stimulation of plasminogen activator inhibitor-1 ( P05121 ) promoter activity by TGF-beta1 from 11.5- to 4-fold and 4.9-fold , respectively . Similar results were observed with the type I collagen promoters . TGF-beta1 increased P27361 activity 4.5-fold at 5 min and 3 . 1-fold at 3 h , while Jun kinase and p38 activity were not affected . Cotransfection of a dominant negative mutant of the small G protein , Rac , but not dominant negative Ras , Cdc42 , or Rho mutants , reduced the effects of TGF-beta1 on the P05121 promoter by approximately half . In support of a role for Rac in signaling by TGF-beta , GTP binding to Rac was increased 3.7-fold following exposure of NIH 3T3 cells to TGF-beta1 for 3 min . These findings indicate that TGF-beta1 modulates gene expression partly through P29323 and Rac in NIH 3T3 cells . Angiotensin II and basic fibroblast growth factor mitogenic pathways in human fetal mesangial cells . Angiotensin II ( Ang II ) and basic fibroblast growth factor ( P09038 / P09038 ) play relevant roles in renal development . Since the signaling pathways modulating the mitogenic effects of Ang II and P09038 in human fetal mesangial cells ( HFMc ) are not clearly defined , we carried out experiments to determine whether they would exert their mitogenic effects by modulating the activity of the mitogen-activated protein kinases ( MAPK ) [ extracellular signal-regulated kinase-2 ( P28482 ) ] and DB02527 signaling pathways . In confluent HFMc , P09038 ( 20 ng/mL ) induced a significant 4-fold increase in P28482 activity and [ 3H ] -thymidine incorporation ( 6-fold ) . In contrast , under similar tissue culture conditions , Ang II ( 10(-6) M ) induced a more modest increase in P28482 activity ( 2-fold ) and [ 3H ] -thymidine incorporation ( 35 +/- 4 % ) . The mitogen-activated protein kinase kinase-1 ( MEK-1 ) inhibitor PD098059 ( 25 microM ) almost completely abolished the P09038 -induced proliferation in HFMc but did not significantly affect Ang II proliferative effects . In the presence of the DB02527 elevating agent isoproterenol , Ang II and P09038 induced opposite changes in DB02527 accumulation and cell growth . DB01064 inhibited the basal and P09038 -induced proliferation of HFMc through a MEK-1/2-independent pathway that included the accumulation of DB02527 . In contrast , isoproterenol increased Ang II mitogenic effects in correlation with a reduction in DB02527 accumulation . We conclude that Ang II and P09038 modulate the proliferation of HFMc through the stimulation of different MEK-1/2-dependent and independent signaling pathways . Activation of MEK-1/2 is required but not sufficient for mitogenesis in HFMc . The accumulation of DB02527 in HFMc counteracts the mitogenic effects of P09038 by a MEK-1/2-independent pathway . 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 . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Enhancement of the P11362 signaling in the P11362 - P08908 heteroreceptor complex in midbrain raphe 5-HT neuron systems . Relevance for neuroplasticity and depression . New findings show existence of P11362 - P08908 heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus . Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity . The existence of P11362 - P08908 heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the P09038 / P11362 mechanism in these nerve cells as well . Therefore , the agonist modulation of the P11362 - P08908 heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells . The combined i.c.v. treatment with P09038 and the P08908 agonist 8-OHDPAT synergistically increased P11362 and P27361 /2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells . Cotreatment with P09038 and the P08908 agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity . These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the P08908 receptor . Taken together , the P08908 autoreceptors by being part of a P11362 - P08908 heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons . Downregulation of angiotensin converting enzyme II is associated with pacing-induced sustained atrial fibrillation . Atrial fibrillation ( AF ) , the most common cardiac arrhythmia , is frequently accompanied by atrial interstitial fibrosis . Angiotensin II ( Ang II ) dependent signaling pathways have been implicated in interstitial fibrosis during the development of AF . However , Ang II could be further degraded by angiotensin converting enzyme II ( Q9BYF1 ) . We examined expression of Q9BYF1 in the fibrillating atria of pigs and its involvement in fibrotic pathogenesis during AF . Nine adult pigs underwent continuous rapid atrial pacing to induce sustained AF and six pigs were sham controls ( i.e. , sinus rhythm ; SR ) . In the histological examinations , extensive accumulation of extracellular matrix in the interstitial space of the atria , as evidenced by Masson 's trichrome stain , were found in fibrillating atria . The relative amount of collagen type I in the atria with AF was significantly increased as compared with that in the SR . Local P12821 activity in the fibrillating atria was also markedly higher than that in the SR subjects . Q9BYF1 gene and protein expression in the AF subjects were significantly decreased compared with those in the SR subjects , whereas expression of mitogen-activated/ P29323 kinase 1/2 ( Q02750 /2 ) , extracellular signal-regulated protein kinase 2 ( P28482 ) , and activated P28482 were significantly greater in the AF subjects . We propose that decreasing Q9BYF1 expression during AF may affect the Ang II-dependent signaling pathway . In addition , our results suggest that atrial fibrosis in AF may be induced by antagonistic regulation between P12821 and Q9BYF1 expression . The use of IRES-based bicistronic vectors allows the stable expression of recombinant G-protein coupled receptors such as NPY5 and histamine 4 . Stable expression of G protein coupled receptors in cell lines is a crucial tool for the characterization of the molecular pharmacology of receptors and the screening for new antagonists . However , in some instances , many difficulties have been encountered to obtain stable cell lines expressing functional receptors . Here , we addressed the question of vector optimization to establish cell lines expressing the human neuropeptide Y receptor 5 ( Q15761 ) or histamine receptor 4 ( Q9H3N8 ) . We have compared bicistronic vectors containing viral or cellular internal ribosome entry sites ( IRES ) , co-expressing the receptor and the neomycine resistance gene from a single mRNA , to a bigenic vector containing two distinct promoters upstream each different genes . This study is the first one to validate the use of three cellular IRESs for long-term transgene expression . Our results demonstrate for both Q15761 and Q9H3N8 that the bicistronic vectors with EMCV , P15692 , FGF1A or P09038 IRES provide clones expressing functional receptors with yields between 25 % and 100 % . In contrast , the bigenic vector provided no functional clones , related to a low expression of Q15761 mRNA . The cell lines expressing active receptor were stable after more than 50 passages . These data indicate that IRES-based bicistronic vectors are particularly appropriate to establish cell clones expressing active G-coupled protein receptors with a high yield . In the case of NPY5 , it was a new way to produce such a stable cell line . Furthermore , the characteristics-presented herein-of this receptor pharmacological property are perfectly in line with those reported in the literature . Ex vivo binding of flibanserin to serotonin P08908 and 5- Q13049 receptors . DB04908 has been reported to be an agonist at P08908 -receptors and an antagonist at 5- Q13049 receptors , with higher affinity for P08908 receptors . Despite the fact that less receptor occupation is required by full agonists than by antagonists to exert their effects , flibanserin was shown to exert 5- Q13049 antagonism at doses ( 4-5 mg kg-1 ) that are lower or equal to those required to stimulate P08908 receptors . In order to understand this phenomenon , the interaction of flibanserin with P08908 and 5- Q13049 receptors was evaluated in ex vivo binding studies . This interaction was evaluated in the prefrontal cortex , hippocampus and midbrain by using [3H]8-OH-DPAT and [3H]ketanserin to label P08908 and 5- Q13049 receptors , respectively . DB04908 was given at 1 , 10 and 30 mg kg-1 intraperitoneally . The dose of 1 mg kg-1 displaced both radioligands preferentially in the frontal cortex . The doses of 10 and 30 mg kg-1 reduced the binding of both radioligands in all the three brain regions non-selectively by about 50 % and 70 % , respectively . The displacement was maximal after 0.5 h and was reduced or not evident after 3 h . We conclude that 5-HT2 antagonism brought about by low doses of flibanserin may reflect functional mechanisms more than receptor-mediated effects . P00797 inhibition with aliskiren . 1. Initial attempts to inhibit renin in humans have faced numerous difficulties . Molecular modelling and X-ray crystallography of the active site of renin have led to the development of new orally active renin inhibitors , such as aliskiren . 2 . DB09026 has a low bioavailability ( between 2.6 and 5.0 % ) compensated by its high potency to inhibit renin ( IC50 : 0.6 nmol/L ) and a long plasma half-life ( 23-36 h ) , which makes it suitable for once-daily dosing . 3 . The once-daily administration of aliskiren to hypertensive patients lowers BP as strongly as standard doses of established angiotensin II type 1 ( AT1 ) receptor blockers ( losartan , valsartan , irbesartan ) , hydrochlorothiazide , angiotensin converting enzyme inhibitors ( ramipril and lisinopril ) or long acting calcium channel blockers ( amlodipine ) . In combination therapy , aliskiren further decreases blood pressure when combined with either hydrochlorothiazide , amlodipine , irbesartan or ramipril . 4 . The biochemical consequences of renin inhibition differ from those of angiotensin I-converting enzyme ( P12821 ) inhibition and Ang II antagonism , particularly in terms of angiotensin profiles and interactions with the bradykinin-nitric oxide-cyclic guanosine monophosphate pathway and possibly the (pro)renin receptor . 5 . Blockade of the renin angiotensin system ( DB01367 ) with P12821 inhibitors , AT1 receptor blockers or a combination of these drugs has become one of the most successful therapeutic approaches in medicine . However , it remains unclear how to optimize DB01367 blockade to maximize cardiovascular and renal benefits . In this context , renin inhibition to render the DB01367 fully quiescent is a new possibility requiring further study .

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How does almotriptan compare with other triptans ? A review of data from placebo-controlled clinical trials . DB00918 , the new selective P28222 /1D agonist , has a higher oral bioavailability than any other DB00669 , with more than two thirds of the administered dose absorbed within the first hour both inside and outside of a migraine attack . Gender or the presence of food in the stomach does not affect its pharmacokinetic profile , and the compound has no clinically relevant interactions with other drugs . Among the available triptans , response rates at 2 hours range from 50 % to 80 % , with 20 % to 50 % of patients pain-free . DB00918 12.5 mg provides similar efficacy , with significant advantage over placebo at 30 minutes and a reliable consistency ( 75 % in two of three attacks ) . Headache typically recurs in 25 % to 45 % of patients with most triptans . The recurrence rate with almotriptan 12.5 mg , 18 % to 27 % , is among the lowest reported . The tolerability of almotriptan 12.5 mg is close to that of placebo with a low incidence of central nervous system side effects and chest symptoms . In conclusion , almotriptan 's consistent pharmacokinetics and good efficacy , in combination with excellent tolerability , make it an attractive choice in the acute treatment of migraine attacks . DB00918 for the acute treatment of adolescent migraine . IMPORTANCE OF THE FIELD : Migraine is a common problem affecting 10 - 20 % of adolescents . Its treatment has three fundamental components : bio-behavioral interventions , preventive measures , and acute drug therapy . In June 2009 , the US FDA approved almotriptan , a P28222 /1D receptor agonist , for the acute treatment of migraine in adolescents aged 12-17 years -- the first ' DB00669 ' to be approved for adolescents . AREAS COVERED IN THIS REVIEW : This review will provide an overview of migraine in adolescents focusing on epidemiology , pathophysiology , classification and a review of treatment options with attention on the evidence from the past 5 years surrounding almotriptan . WHAT THE READER WILL GAIN : Given its recent FDA approval , it is important to for clinicians and pharmacists to become familiar with the clinical spectrum of migraine in teenagers and with recent evidence on this newly approved agent , almotriptan . TAKE HOME MESSAGE : DB00918 is a safe , effective , and approved agent for the acute treatment of migraine headache in adolescents . Emergence of motor circuit activity . In the developing nervous system , ordered neuronal activity patterns can occur even in the absence of sensory input and to investigate how these arise , we have used the model system of the embryonic chicken spinal motor circuit , focusing on motor neurons of the lateral motor column ( O15467 ) . At the earliest stages of their molecular differentiation , we can detect differences between medial and lateral O15467 neurons in terms of expression of neurotransmitter receptor subunits , including P30532 , P36544 , Q12879 , P39086 , P08908 and P28222 , as well as the Q9H2X9 transporter . Using patch-clamp recordings we also demonstrate that medial and lateral O15467 motor neurons have subtly different activity patterns that reflect the differential expression of neurotransmitter receptor subunits . Using a combination of patch-clamp recordings in single neurons and calcium-imaging of motor neuron populations , we demonstrate that inhibition of nicotinic , muscarinic or GABA-ergic activity , has profound effects of motor circuit activity during the initial stages of neuromuscular junction formation . Finally , by analysing the activity of large populations of motor neurons at different developmental stages , we show that the asynchronous , disordered neuronal activity that occurs at early stages of circuit formation develops into organised , synchronous activity evident at the stage of O15467 neuron muscle innervation . In light of the considerable diversity of neurotransmitter receptor expression , activity patterns in the O15467 are surprisingly similar between neuronal types , however the emergence of patterned activity , in conjunction with the differential expression of transmitter systems likely leads to the development of near-mature patterns of locomotor activity by perinatal ages . Effective dasatinib uptake may occur without human organic cation transporter 1 ( O15245 ) : implications for the treatment of imatinib-resistant chronic myeloid leukemia . We have previously shown that imatinib uptake into chronic myeloid leukemia ( CML ) cells is dependent on human organic cation transporter 1 ( O15245 ; O15245 ) , and that low O15245 expression is an important determinant of clinical outcome to imatinib treatment . We hypothesized that dasatinib might be transported differently than imatinib , possibly accounting for its favorable effects in imatinib-resistant patients . (14)C-dasatinib uptake was greater in KCL22-transfected cells with pcDNA3- O15245 plasmid ( high O15245 -expressing cells ) than in control cells ( P = .02 ) . However , hOCT inhibitors did not decrease dasatinib uptake into either control or primary cells , in contrast to their block on imatinib uptake . Dasa-tinib decreased the level of phosphorylated CrkL to 49.9 % in control and 40.3 % in high O15245 -expressing cells . Dasa-tinib efflux was investigated in confluent P08183 -transfected MDCKII cell monolayers . Both dasatinib and imatinib were transported from the basal to the apical layer , indicating that they were transported by P08183 , which was confirmed using the P08183 inhibitor PSC833 ( P = .001 and P < .001 , respectively ) . Compared with imatinib , dasatinib achieved superior intracellular levels and P11274 - P00519 suppression even in cells with low or blocked O15245 . Efflux of dasatinib and imatinib appear similar via P08183 . Dasatinib may therefore offer an advantage over imatinib in patients with low O15245 expression . Possible involvement of endocannabinoids in the increase of morphine consumption in maternally deprived rat . Whether adolescent exposure to chronic DB00470 ( THC ) facilitates progression to opioid consumption is still controversial . In a maternal deprivation model ( 3 h daily from postnatal day 1-14 ) , we previously reported that adolescent exposure to chronic THC blocks morphine dependence in maternally deprived ( D ) rats . Owing to the existence of a functional cross-interaction between the opioid and cannabinoid systems in reward , we evaluated if the vulnerability to opiate reward in D rats , may involve an alteration of the endocannabinoid system . Anandamide and 2-arachidonoylglycerol ( 2-AG ) , were quantified in the striatum and mesencephalon of adolescent and adult D and non-deprived ( animal facility rearing , AFR ) rats by isotope dilution liquid chromatography-mass spectrometry . Oral morphine self-administration behavior was analyzed for 14 weeks , 24 days after chronic injection of the cannabinoid P21554 receptor antagonist/inverse agonist , SR141716A ( 3 mg/kg ) for 2 weeks during adolescence ( P01160 35-48 ) . Adolescent D rats exhibited higher basal levels of anandamide than adolescent AFR rats in the nucleus accumbens ( 38 % ) , the caudate-putamen nucleus ( 62 % ) and the mesencephalon ( 320 % ) , whereas adult D rats showed an increase of anandamide and 2-AG levels in the nucleus accumbens ( 50 % and 24 % , respectively ) and of 2-AG in the caudate-putamen nucleus ( 48 % ) , compared to adult AFR rats . Chronic administration of SR141716A to adolescent D rats blocked the escalation behavior in the morphine consumption test . Our data suggest that altered brain endocannabinoid levels may contribute to the escalation behavior in the morphine consumption test in a maternal deprivation model . The pipeline in headache therapy . Migraine is a common , disabling , neurovascular disorder characterized by episodic attacks of head pain and associated disability plus systemic autonomic and neurologic symptoms . The advent of the DB00669 class of medication in the 1990s revolutionized the acute treatment of migraine , but many migraineurs do not respond optimally or at all to triptans , have intolerable adverse effects , or have contraindications to their use . Preventive pharmacotherapy has advanced mostly through serendipity , with new drugs being found effective while being used for other indications . There remains a significant need for new medications and devices that can provide effective , rapid , and sustained pain relief without adverse effects or recurrence . Several new acute and preventive therapies for the treatment of migraine and cluster headaches have shown promise and are currently under investigation . This article covers innovative delivery mechanisms , calcitonin gene-related peptide receptor antagonists , antibodies to calcitonin gene-related peptide and its receptor , P30939 receptor agonists , transient receptor potential vanilloid receptor modulators , orexin receptor antagonists , glial cell modulators , and neurostimulation . The pharmacological profile and clinical prospects of the oral P30939 receptor agonist lasmiditan in the acute treatment of migraine . More than 20 years have passed without the launch of a new substance class for acute migraine therapy . Triptans were the latest class of substances which successfully passed all developmental stages with a significant antimigraine efficacy and a sufficient safety profile . New drugs with a better adverse event profile and at least similar efficacy are needed for migraine subjects who can not tolerate triptans for attack treatment . Lasmiditan is a novel highly specific P30939 receptor agonist currently in clinical trials for acute migraine therapy and devoid of vasoconstriction in coronary arteries as determined in a surrogate assay . In both phase II randomized , placebo-controlled trials in acute migraine the primary endpoint was met . For the intravenous formulation a clear dose-dependent effect on headaches could be determined . Lasmiditan tablets in doses of 50-400 mg show significant headache relief after 2 hours compared with placebo and improved accompanying symptoms . This substance is chemically clearly different from other antimigraine drugs , which is also reflected by its dose-dependent adverse event profile chiefly including dizziness , vertigo , paresthesia and fatigue . Adverse events are usually linked to the central nervous system . Future phase III clinical trials with an active DB00669 comparator or in a preferential trial design will allow a better comparison of lasmiditan and triptans . They will also determine whether lasmiditan will become available to the migraine patient . Serotonergic modulation of the acoustic startle response in rats during preweaning development . The involvement of serotonin ( 5-HT ) in modulating the acoustic startle response ( ASR ) is well established in adult rats , but 5-HT involvement during the preweaning period , when 5-HT neurons undergo extensive development , has not previously been described . Three 5-HT receptor subtypes are reported to modulate the ASR in adult rats : P08908 and 5-HT2 receptor agonists facilitate the ASR , whereas P28222 agonists decrease the response . In the present study , the effects of 5-HT agonists and generalized 5-HT depletion on the ASR were studied in preweanling animals , using independent groups of Long-Evans rats tested on postnatal day ( P01160 ) 13 , 17 and 21 . 8-Hydroxy-2-(di-n-propylamino) tetralin ( 8OHDPAT , 62-1000 micrograms/kg ) , a P08908 receptor agonist , and 5-methoxy-N,N-dimethyl tryptamine ( MeODMT , 2-4 mg/kg ) , a nonselective 5-HT agonist , had no effect on P01160 13 and then increased the ASR on P01160 17 and 21 . The 5-HT2 receptor antagonists cyproheptadine ( 5 mg/kg ) and ketanserin ( 5 mg/kg ) blocked the effect of MeODMT at both ages , providing some evidence that MeODMT increased the ASR through 5-HT2 receptors . 1-(m-Chlorophenyl) piperazine ( mCPP , 1-5 mg/kg ) , a P28222 agonist , had no effect on ASR amplitude on P01160 13 or 17 and then produced a dose-related decrease in the response on P01160 21 . Generalized depletion of 5-HT by 80-90 % in whole-brain and spinal cord , using p-chlorophenylalanine ( PCPA , 300 mg/kg 24 hr prior to testing ) , did not alter ASR amplitude at any age. ( ABSTRACT TRUNCATED AT 250 WORDS ) [ Pathophysiological basis of 3 subtypes in ganfeng neidong syndrome ] . The multiple parameters of 3 Subtypes : Ganyang Huafeng Syndrome ( GYHFS ) , Xuexu Shengfeng Syndrome and Yinxu Fengdong Syndrome of Ganfeng Neidong Syndrome were determined for the 1st time . It was found that there were several characteristics in GYHFS . ( 1 ) Disturbance of the cerebral blood flow and the damage of brain tissue was manifested by the abnormality of the bulbar conjunctival microcirculation , carotid Doppler ultrasonic determination and brainstem auditory and visual pathway , high blood viscosity , dysmnesia , free radical and lipid peroxidation injury and the changes of Zn , Cu , K and Mg after brain damage . ( 2 ) Stress status were expressed by the high plasma levels of cortisol , norepinephrine and epinephrine , decreased serum triiodothyronine level and hyperfunction of sympathetic nerve . ( 3 ) The marked changes of the regulating substance of the vessel smooth muscle function including the increased plasma levels of TXB2 , TXB2/6-k-PGF1 alpha , and calmodulin , as well as decreased SP , P01160 , P80511 . Other 2 subtypes had about the same changes of these parameters , but of milder disorders . Genomic organization , expression and evolution of porcine Q15648 , 2 , and 3 . Recently we identified and characterized porcine calcitonin receptor-stimulating peptide ( CRSP ) 1 , O60244 and Q9ULK4 as members of the calcitonin/calcitonin gene-related peptide ( CT/ P80511 ) family . In the present study , the genomic sequences and organization of Q15648 , 2 , and 3 were determined , and the expression of the genes in the porcine brain was investigated using in situ hybridization . Analysis of 5'-upstream regions of the three CRSPs demonstrated that Q15648 and O60244 have almost identical sequences ( > 98 % similarity ) and high sequence similarities including functional transcription binding sites with the corresponding region of human CALCA ( CT/alpha P80511 ) , whereas Q9ULK4 retains less similarity with the above genes . RH mapping of CRSPs demonstrated that they resided in a region of swine chromosome 2 ( Q9NXB9 ) . The arrangement of the genes in the region was found to be conserved in corresponding human and mouse regions . In situ hybridization demonstrated sense transcripts of the three genes in cerebrum , hippocampus , hypothalamus , pons/midbrain , and thalamus of 3-month-old pigs , and O60244 sense transcripts additionally in tractus opticus . The sense transcripts of alpha P80511 and P10092 ( beta P80511 ) were detected in cerebrum , hippocampus , and pons/midbrain of newborn mice , and to a lesser extent in pons/midbrain of 8-week-old mice . These results taken together with the chromosomal conservation and phylogenetic clustering of CT/ P80511 family indicate that Q15648 , 2 , and 3 may be functionally different from alpha P80511 and beta P80511 , though they are indicated to have a common progenitor gene . The search for novel migraine therapies : experimental models . The identification and development of the potent P28222 /1D agonist , sumatriptan has resulted in new therapeutic opportunities for the treatment of migraine and a number of chemically novel agents with a similar mechanism of action have been identified . Whilst these agents are optimised to enhance the therapeutic effect of sumatriptan , development of mechanistically novel therapies may provide new directions for the care of migraine sufferers . To develop new treatment paradigms , novel chemical entities should be evaluated in animal models which are predictive of therapeutic efficacy e.g. : in animal models where sumatriptan has shown activity , or the pathophysiological processes involved in the disease must be targeted . Therefore , investigation of mechanisms underlying cortical activity and its involvement in the activation of trigeminal vascular pathways may allow better understanding of the disease and result in the identification of new non- DB00669 -like therapies . Enhancement of cytotoxicity of natural product drugs against multidrug resistant variant cell lines of human head and neck squamous cell carcinoma and breast carcinoma by tesmilifene . N,N-diethyl-2-[4-(phenylmethyl)phenoxyl]ethanamine ( tesmilifene ) , a tamoxifen derivative with antihistamine activity , greatly enhanced the survival of doxorubicin-treated , advanced stage breast cancer patients in a phase III trial . However , the molecular basis of tesmilifene action is not firmly established . The effects of tesmilifene on activity of several anticancer drugs was investigated using human head and neck squamous cell carcinoma ( HNSCC ) and breast carcinoma cell lines as a model system . Multidrug resistant ( MDR ) variants of an HNSCC cell line , HN-5a/V15e , and a breast carcinoma cell line , MCF-7/V25a , both highly overexpressed mdr1 ( P08183 ) mRNA and the proteins P-glycoprotein and glutathione transferase-pi . Drug sensitivities were measured by a vital stain after 4 days of continuous exposure to anticancer drug in the absence and presence of tesmilifene at a concentration that alone had no antiproliferative effect . DB04905 had minimal effect on drug cytotoxicity against the parental cell lines . However , the same tesmilifene treatment enhanced cytotoxicity of docetaxel , paclitaxel , epirubicin , doxorubicin , and vinorelbine against both MDR cell lines by up to 50 % . Flow cytometric measurement of annexin V/propidium iodide staining demonstrated that tesmilifene increased the killing of HN-5a/V15e cells caused by docetaxel after 24 and 48h exposure . DB04905 increased accumulation of radiolabelled vincristine in HN-5a/V15e cells , over 4h , by up to 100 % . The results suggest that tesmilifene might be effective in the treatment of tumors that are resistant to natural product drugs . The mechanism of enhancement appears to be related to expression of an ABC pump-dependent , MDR phenotype . Development and evaluation of high throughput functional assay methods for Q12809 potassium channel . Three functional hERG channel assay methods have been developed and evaluated . The methods were tested against five known hERG channel inhibitors : dofetilide , terfenadine ( Seldane ) , sertindole ( DB06144 ) , astemizole ( Hismanal ) , and cisapride ( Propulsid ) . The DiBAC4(3)-based assays were found to be the most economical but had high false-hit rates as a result of the interaction of dye with the test compounds . The membrane potential dye assay had fewer color-quenching problems but was expensive and still gave false hits . The nonradioactive Rb+ efflux assay was the most sensitive of all the assays evaluated and had the lowest false-hit rate . [ The cluster headache : a clinical model of immunologic receptor pathology ? ] . It is well established that cluster headache shows impaired functions at their neuroimmunomodulatory system level . Defect in receptor expression for 5-HT , IL-1 and P60568 have been found in these patients . Sumatriptan , a molecule with agonistic activity for P28221 receptor , truncates cluster headache attacks in 74 % of patients . Flow cytometric analysis of monocytes expressing 5-HT receptor in cluster headache patients showed different trends clearly correlated with the clinical response to sumatriptan . Our findings strongly support the concept that cluster headache patients who are non responders to sumatriptan could present a block in their 5-HT receptor possibly due to specific autoantibodies for this receptor site . P03372 -immunoreactive neurons contain calcitonin gene-related peptide , methionine-enkephalin or tyrosine hydroxylase in the female rat preoptic area . We have shown in our previous studies that estrogen treatment selectively influences calcitonin gene-related peptide ( P80511 ) - , methionine-enkephalin ( DB00134 -Enk ) - and tyrosine hydroxylase ( TH ) -immunoreactive ( IR ) intensities in the neurons of the periventricular preoptic nucleus ( Q9H237 ) and the medial preoptic area ( DB00603 ) of the female rat . In the present study , we examined whether estrogen receptor ( ER ) -IR neurons in the Q9H237 and DB00603 contain P80511 , DB00134 -Enk , or TH using a double-labeling immunohistochemical method and investigated changes in the number of double-labeling cells upon treatment with estrogen . Brain sections of ovariectomized rats and ovariectomized and estrogen-treated rat were stained using the avidin-biotin-peroxidase complex method followed by the peroxidase-anti-peroxidase method . The sections were first incubated with an anti-ER antibody in conjunction with nickel diaminobenzidine which produces a dark blue reaction product in the nucleus . Subsequently , P80511 , DB00134 -Enk or TH antisera were applied to these sections and the resulting brown diaminobenzidine reaction product in the cytoplasm was examined . Neurons that were double-labeled for ER and P80511 , DB00134 -Enk or TH were investigated in the Q9H237 and DB00603 . The number of doubly labeled ER/ P80511 - and ER/TH-IR neurons was large , whereas the number of ER/ DB00134 -Enk-IR neurons was small . These results suggest that ER in the Q9H237 and DB00603 may be more closely related to the mechanism of changes in P80511 - and TH-IR intensities upon estrogen treatment than that in DB00134 -Enk-IR intensity . [ Role of serotonin and other neuroactive molecules in the physiopathogenesis of migraine. Current hypotheses ] . The study of the mechanisms of action of the DB00669 group of drugs has largely contributed to the progress made in the understanding of the physiopathological processes that are possibly responsible for migraine . In this context , two discoveries have been especially important : 1 ) these anti-migraine drugs are specifically recognized by three main types of serotonin receptors ( P28222 , P28221 , and P30939 ) ; and 2 ) these receptors are present in the meninges , where they are expressed by both smooth muscle cells and/or endothelial cells of the vascular wall and/or the perivascular trigeminal to be deleted axon terminals . These two findings have led to the most currently accepted physiopathogenic hypothesis , whereby the migraine attack would start with an excitation of the perivascular trigeminal to be deleted fibers , which would then trigger the release of vasoactive peptides ( DB05875 , calcitonin gene-related peptide/ P80511 ) within the dura mater . Locally , i.e. , in the dura mater in particular , these substances can provoke vasodilatation ( P80511 ) and plasmatic extravasation ( DB05875 ) with platelet lysis and mast cell degranulation , thereby leading to the release of algogenic substances that excite the neighboring trigeminal fibers , and this neurogenic inflammatory response can progressivelly extend to the meninges as a whole . This reaction subsequently reaches the bulbar and thalamic nuclei and then the sensory cortex , where it is integrated and expressed as migraine pain . The aim of this article was to report the main findings on endogenous substances ( serotonin , peptides , nitric oxide [ NO ] , etc. ) which appear to play a key role in this physiopathogenic sequence . Increased expression of receptor for advanced glycation end products by synovial tissue macrophages in rheumatoid arthritis . OBJECTIVE : The accumulation of advanced glycation end products ( AGEs ) , P80511 , and high mobility group box chromosomal protein 1 has been associated with joint inflammation in rheumatoid arthritis ( RA ) . This study was undertaken to determine the induction of the receptor for these proteins , termed receptor for AGEs ( RAGE ) , in synovial tissue ( ST ) macrophages from RA patients . METHODS : RAGE and P34810 expression in ST were determined by 2-color immunofluorescence labeling . Cell surface and messenger RNA ( mRNA ) expression of RAGE were examined by flow cytometry and reverse transcriptase-polymerase chain reaction ( PCR ) or real-time PCR , respectively . RESULTS : P34810 + lining macrophages , like the vasculature , expressed high levels of RAGE in inflamed ST from RA patients . RAGE mRNA expression was significantly higher in RA ST than in ST from patients with osteoarthritis . RAGE mRNA levels were significantly higher in ST macrophages and normal endothelial cells than in ST P01730 + T cells and synovial fibroblasts stimulated with tumor necrosis factor alpha and interleukin-1beta ( IL-1beta ) . Cell surface RAGE was highly induced on normal monocytes after a 24-hour incubation with a 20 % concentration of RA ST cell culture supernatants . RAGE mRNA expression in adherent monocytes was augmented by various cytokines , most potently by IL-1beta . CONCLUSION : These results indicate that RAGE overexpression in lining macrophages may be induced , at least in part , by cytokines such as IL-1 , leading to the amplification of inflammatory responses mediated by RAGE ligands that are abundant in RA joints . [ Effects of P80511 on the P12830 expression in human bronchial epithelial cells ] . OBJECTIVE : To discuss the effect of calcitonin gene-related peptides ( P80511 ) on epithelial cadherin ( E-cd ) expression in human bronchial epithelial cells ( HBECs ) in vitro . METHODS : The effect of P80511 on E-cd protein and mRNA expression in both normal and O3-challenged HBECs were determined by immunocytochemistry and RT-PCR . The signal transduction pathways of P80511 were observed by using protein kinase C(PKC) inhibitor ( H-7 ) , calmodulin( P62158 ) inhibitor ( W-7 ) and PKA inhibitor ( H-89 ) . RESULTS : P80511 increased E-cd mRNA and protein expressions of normal and O3-challenged HBECs in a dose-dependent manner . P80511 had no effect on cytoplasm E-cd expression . Pre-treatment with H-89 , H-7 and W-7 , the up-regulatory effect of P80511 on E-cd expression was partly abolished . CONCLUSION : P80511 increased in cytomembrane E-cd expression of normal and O3-challenged HBECs in a dose-dependent manner . E-cd expression on HBECs was strengthened by P80511 via PKA , PKC and P62158 pathways . Rare human nicotinic acetylcholine receptor α4 subunit ( P43681 ) variants affect expression and function of high-affinity nicotinic acetylcholine receptors . DB00184 , the primary psychoactive component in tobacco smoke , produces its behavioral effects through interactions with neuronal nicotinic acetylcholine receptors ( nAChRs ) . α4β2 nAChRs are the most abundant in mammalian brain , and converging evidence shows that this subtype mediates the rewarding and reinforcing effects of nicotine . A number of rare variants in the P43681 gene that encode the α4 nAChR subunit have been identified in human subjects and appear to be underrepresented in a cohort of smokers . We compared three of these variants ( α4R336C , α4P451L , and α4R487Q ) to the common variant to determine their effects on α4β2 nAChR pharmacology . We examined [(3)H]epibatidine binding , interacting proteins , and phosphorylation of the α4 nAChR subunit with liquid chromatography and tandem mass spectrometry ( LC-MS/MS ) in P29320 293 cells and voltage-clamp electrophysiology in Xenopus laevis oocytes . We observed significant effects of the α4 variants on nAChR expression , subcellular distribution , and sensitivity to nicotine-induced receptor upregulation . Proteomic analysis of immunopurified α4β2 nAChRs incorporating the rare variants identified considerable differences in the intracellular interactomes due to these single amino acid substitutions . Electrophysiological characterization in X. laevis oocytes revealed alterations in the functional parameters of activation by nAChR agonists conferred by these α4 rare variants , as well as shifts in receptor function after incubation with nicotine . Taken together , these experiments suggest that genetic variation at P43681 alters the assembly and expression of human α4β2 nAChRs , resulting in receptors that are more sensitive to nicotine exposure than those assembled with the common α4 variant . The changes in nAChR pharmacology could contribute to differences in responses to smoked nicotine in individuals harboring these rare variants . [ Migraine - established concepts and new developments ] . Migraine is a very common primary headache disorder associated with intermittent attacks and great suffering . Despite extensive research efforts in the recent years , many pathophysiological aspects remain unclear . An altered cortical adaptability and the brainstem as a migraine generator are probably involved in the initiation of a ( silent ) cortical spreading depression and other processes that lead to neurogenic inflammation of the meninges causing the headache . Numerous studies in the last years have examined somatic , especially cerebrovascular and also psychological comorbidities . For attack therapy , P80511 antagonists have emerged as promising non-vasoconstrictive acting alternatives for triptans . However , they were so far not approved due to liver enzyme elevations in safety studies . Another new approach without vasoconstrictive action are the selective P30939 agonists ( especially Lasmiditan ) . Large placebo-controlled and DB00669 -controlled trials need to be awaited . For migraine prophylaxis , a comparable effect of sports and pharmacological prophylaxis using topiramate could be found . Particularly the combination of drug and non-drug therapies ( such as the combination of stress management training with a beta-blocker treatment ) achieves high efficacy . Also interdisciplinary , multimodal treatment approaches are important options . Two large multicentre studies have demonstrated the efficacy of botulinum toxin A as a prophylactic treatment for chronic migraine . Neuromodulative and neurostimulative procedures are promising but still experimental treatment options for patients with refractory migraine . Curcumin inhibits the metastasis of P04264 papillary thyroid cancer cells via modulating P12830 and matrix metalloproteinase-9 expression . The anti-metastatic effect of curcumin on papillary thyroid cancer P04264 cells and its underlying mechanisms were investigated . Curcumin at 12.5 , 25 and 50 μM promoted mesenchymal-epithelial transition and decreased the migration rate of P04264 cells by 24-87 % . Its mechanism may involve the up-regulation of P12830 expression levels and down-regulation of the activity and expression of matrix metalloproteinase-9 . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . Migraine in the DB00669 era : progresses achieved , lessons learned and future developments . Triptans , serotonin P28222 /1D receptor agonists , more than revolutionizing the treatment of migraine , stimulated also ground breaking research that provided insights into the anatomy , physiology , and molecular pharmacology of migraine . This knowledge , in turn , is stimulating research on new mechanisms of action for the treatment of migraine . Accordingly , it is opportune to critically review the main advances in migraine science that happened in the DB00669 era . Herein we first review and conceptualize some of the progresses achieved in migraine science during the DB00669 era . We then review the class of the triptans -- mechanism of action and clinical evidence . We close by briefly discussing the class of P80511 receptor antagonists , which is currently being developed for the acute treatment of migraine . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Small-dose capsaicin reduces systemic inflammatory responses in septic rats . We investigated the influence of small- and large-dose capsaicin in modulating systemic inflammatory responses during different stages of sepsis in rats . Rats were divided into six groups : group C , control ; group S , sepsis ; group CLC , small dose of capsaicin ( 1 mg/kg subcutaneously ) ; group O00585 , small dose of capsaicin plus sepsis ; group CHC , large dose of capsaicin ( 150 mg/kg subcutaneously ) ; group P29353 , large dose of capsaicin plus sepsis . Rats were made septic by cecal ligation and puncture ( CLP ) . Each group was subdivided into two subgroups . The animals were killed at 9 or 18 h after CLP . Plasma concentrations of calcitonin gene-related peptide ( P80511 ) , tumor necrosis factor ( P01375 ) -alpha , interleukin ( IL ) -6 , P22301 , and total nitrite/nitrate ( NOx ) were measured . Superoxide dismutase and malondialdehyde ( MDA ) were determined in liver , lung , and heart tissues . P80511 was increased in groups S , CLC , and O00585 when compared with the other groups . In the O00585 group , plasma concentrations of P01375 , P05231 , NOx , and tissue MDA levels were reduced and P22301 level was increased when compared with groups S and P29353 18 h after CLP ( P < 0.05 ) . Small-dose capsaicin treatment increased antiinflammatory P22301 levels and attenuated the increases in proinflammatory cytokines , NOx , and tissue MDA in septic rats . Post- DB00669 era for the treatment of acute migraine . There now is one realized and several attractive targets for the treatment of acute attacks of migraine that will follow and augment the use of serotonin P28222 /1D receptor agonists , the triptans . P01258 gene-related peptide ( P80511 ) receptor blockade recently has been shown to be an effective acute antimigraine strategy ; therefore , blockade of P80511 release by inhibition of trigeminal nerves would seem a logical approach . A number of targets are reviewed in this article including serotonin P30939 and P28221 receptors , adenosine A1 receptors , nociceptin , vanilloid Q8NER1 receptors , and anandamide P21554 receptors . Development of one or more such compound offers the exciting prospect of new non-vasoconstrictor treatments for migraine and cluster headache . Triptan use preceding life-threatening arrhythmias in charcot-marie-tooth : a case report and review of the literature . Charcot-Marie-Tooth ( CMT ) identifies a rare group of inherited disorders of the peripheral nervous system that share clinical features of sensory and motor defects , but rarely affect cardiac function . The few references that relate CMT to cardiac pathology of any sort are so rare that their finding was considered either fortuitous or suggestive of an erroneous diagnosis . The P28222 /1D agonists or DB00669 drug class was introduced to the clinical practice arena in the early 1990s , and since then several cardiac adverse events have been associated with its use . The authors report the case of a 41-year-old white woman with CMT who had been recently prescribed sumatriptan and who presented with sudden loss of consciousness associated with ventricular fibrillation . These findings have been reported in the literature , but the association of DB00669 -induced arrhythmias and degenerative neuropathies remains to be established . The authors propose , through this case report and review , that the relative rarity of CMT coupled with the lack of further investigation of their association with conduction abnormalities might have set the stage for underestimation of the potentially synergistic effect with triptans in their capacity to generate life-threatening arrhythmias . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor . Mouse pluripotent hematopoietic stem cells ( PHSC ) were fractionated based on size and density using counterflow centrifugal elutriation ( CCE ) . These heterogeneous PHSC populations were further enriched by subtraction of cells with lineage-specific markers ( Lin- ) followed by positive sorting for c-kit expression . The cells were characterized for their functional and biochemical properties . We defined a subpopulation of c-kit-positive cells that expressed high numbers of c-kit receptors ( c-kitBR ) . One hundred c-kitBR cells from either low- or higher-density fractions were sufficient to repopulate the lymphohematopoietic system in WBB6F1-W/Wv ( W/Wv ) recipients , whereas no PHSC were found in cells with low ( c-kitDULL ) or no ( c-kitNEG ) c-kit expression . Lin- c-kitBR cells were separated into RhoDULL and RhoBR subsets based on their ability to efflux rhodamine 123 ( Rho ) . The PHSC were concentrated in Lin- c-kitBR RhoDULL cells and the number of Lin- c-kitBR RhoBR cells correlated directly with the number of day 12 colony-forming unit-spleen ( CFU- P28222 ) in each fraction . We were not able to enrich further for PHSC using monoclonal antibodies to the cell-surface markers AA4.1 or P01730 , which have been used by others to isolate PHSC . The small , low-density Lin- c-kitBR subset contained PHSC and few CFU- P28222 . This enabled us to assay PHSC for expression of the flk-2 gene , which encodes a tyrosine kinase receptor present on fetal liver PHSC . Purified RNA from the low-density Lin- c-kitBR subset did not contain flk-2 mRNA . We suggest that AA4.1 , P01730 and flk-2 are expressed as stage-specific markers on PHSC in cell cycle . Characterization of the 3p12.3-pcen region associated with tumor suppression in a novel ovarian cancer cell line model genetically modified by chromosome 3 fragment transfer . The genetic analysis of nontumorigenic radiation hybrids generated by transfer of chromosome 3 fragments into the tumorigenic OV-90 ovarian cancer cell line identified the 3p12.3-pcen region as a candidate tumor suppressor gene ( Q9GZX9 ) locus . In the present study , polymorphic microsatellite repeat analysis of the hybrids further defined the 3p12.3-pcen interval to a 16.1 Mb common region containing 12 known or hypothetical genes : 3ptel- Q9HCK4 - Q9Y6N7 - Q04446 - Q8N3J6 - A8MV65 - Q9UQN3 - P28069 - P30939 - Q9UFW8 - Q8IZM8 - Q5JPI3 - P29320 -3pcen . Seven of these genes , Q9Y6N7 , Q04446 , A8MV65 , Q9UQN3 , Q9UFW8 , Q8IZM8 , and Q5JPI3 , exhibited gene expression in the hybrids , placing them as top Q9GZX9 candidates for further analysis . The expression of all but one ( A8MV65 ) of these genes was also detected in the parental OV-90 cell line . Mutations were not identified in a comparative sequence analysis of the predicted protein coding regions of these candidates in OV-90 and donor normal chromosome 3 contig . However , the nondeleterious sequence variants identified in the transcribed regions distinguished parent of origin alleles for Q9Y6N7 , A8MV65 , Q9UQN3 , and Q9UFW8 and cDNA sequencing of the hybrids revealed biallelic expression of these genes . Interestingly , underexpression of A8MV65 and Q8IZM8 were observed in malignant ovarian tumor samples as compared with primary cultures of normal ovarian surface epithelial cells or benign ovarian tumors , and this occurred regardless of allelic content of 3p12.3-pcen . The results taken together suggest that dysregulation of A8MV65 and/or Q8IZM8 expression may have affected pathways important in ovarian tumorigenesis which was offset by the transfer of chromosome 3 fragments in OV-90 , a cell line hemizygous for 3p . DB01076 induces insulin sensitization in Zucker lean and fatty rats . BACKGROUND : The 3-hydroxy-3-methyl glutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors ( ' statins ' ) have been implicated in preventing new onset type 2 diabetes , whereas the mechanism of this effect is not known . We investigated the effects of an P04035 inhibitor , atorvastatin , on insulin sensitization in Zucker lean and fatty rats . METHODS AND RESULTS : In vivo studies of insulin sensitization were performed in chow fed Zucker lean and fatty rats treated with atorvastatin 50mg/kg/day ( ATORVA_50 ) and results were compared to Zucker lean and fatty rats treated with drug vehicle only ( CONT ) . Additional Zucker lean rats were treated with an intermediate dose of atorvastatin 25mg/kg/day ( ATORVA_25 ) . Treatment with atorvastatin resulted in a dose-dependent improvement in whole body insulin sensitivity in both lean and fatty rats , with an approximately two-fold increase in glucose infusion rate and glucose disposal ( Rd ) in ATORVA_50 versus CONT ( p < 0.01 ) . DB01076 50mg/kg/day resulted in an increase in DB08831 ( 2-DOG ) uptake by skeletal muscles ( approximately two-fold increase in 2-DOG uptake in quadriceps ( p=0.06 ) and gastrocnemius ( p < 0.01 ) ) in lean Zucker rats . P01308 -stimulated phosphorylation of Akt/ P31749 was significantly increased in skeletal muscle of ATORVA_50 versus CONT in both lean and fatty rats . CONCLUSION : DB01076 induces insulin sensitization in Zucker lean and fatty rats . This may be a clinically important pleiotropic effect if confirmed in insulin resistant humans . Individual DB00669 selection in migraine attack therapy . About 6 % of men and 18 % of women suffer migraine attacks . Migraine can induce a great impact in the quality of life of the patient and the costs of medical care and lost productivity can be also high . There are two therapeutic approaches in the treatment of migraine : preventive therapy and acute treatment of migraine attack . Immediate treatment with selective serotonin [ P28222 /1T ] receptor agonists ( so-called triptans ) is the first-line option in the acute treatment of moderate-severe migraine attacks . The introduction in early nineties of triptans was a revolution in migraine therapy and evidences about their efficacy are at present irrefutable . At the moment , there are seven marketed molecules : sumatriptan , rizatriptan , zolmitriptan , eletriptan , naratriptan , almotriptan and frovatriptan . Obviously , every molecule has different pharmacokinetic and pharmacodinamic properties and , moreover , some triptans have several formulations : tablets , dissolvable tablets , nasal and injections . The prescription of one of these seven triptans for a specified patient is based in the drug profile : efficacy , safety , pharmacokinetics and pharmacodynamics . Despite there are a lot of published studies using triptans , no clinical trial has analyzed all the molecules at the same time . Other data to take account in the final prescription are clinical characteristics of the migraine attack and patient characteristics : labour aspects , style of life and the patient medical history . We present a state-of-the-art of the DB00669 selection in treatment of moderate-severe migraine attacks . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Endocannabinoids in the brainstem modulate dural trigeminovascular nociceptive traffic via P21554 and " DB00669 " receptors : implications in migraine . Activation and sensitization of trigeminovascular nociceptive pathways is believed to contribute to the neural substrate of the severe and throbbing nature of pain in migraine . Endocannabinoids , as well as being physiologically analgesic , are known to inhibit dural trigeminovascular nociceptive responses . They are also involved in the descending modulation of cutaneous-evoked C-fiber spinal nociceptive responses from the brainstem . The purpose of this study was to determine whether endocannabinoids are involved in the descending modulation of dural and/or cutaneous facial trigeminovascular nociceptive responses , from the brainstem ventrolateral periaqueductal gray ( vlPAG ) . P21554 receptor activation in the vlPAG attenuated dural-evoked Aδ-fiber neurons ( maximally by 19 % ) and basal spontaneous activity ( maximally by 33 % ) in the rat trigeminocervical complex , but there was no effect on cutaneous facial receptive field responses . This inhibitory vlPAG-mediated modulation was inhibited by specific P21554 receptor antagonism , given via the vlPAG , and with a P28222 /1D receptor antagonist , given either locally in the vlPAG or systemically . These findings demonstrate for the first time that brainstem endocannabinoids provide descending modulation of both basal trigeminovascular neuronal tone and Aδ-fiber dural-nociceptive responses , which differs from the way the brainstem modulates spinal nociceptive transmission . Furthermore , our data demonstrate a novel interaction between serotonergic and endocannabinoid systems in the processing of somatosensory nociceptive information , suggesting that some of the therapeutic action of triptans may be via endocannabinoid containing neurons in the vlPAG . Sumatriptan ( P28221 receptor agonist ) does not exacerbate symptoms in obsessive compulsive disorder . The non-selective serotonin ( 5-HT ) receptor agonist meta-chlorophenylpiperazine ( mCPP ) has been reported to elicit symptoms in patients with obsessive compulsive disorder ( OCD ) . MK-212 , another non-selective 5-HT receptor agonist , does not seem to induce obsessive compulsive symptoms in OCD patients . The major pharmacological difference between mCPP and MK-212 is their affinity for the 5-HT(ID) receptor . The aim of this study was to explore the role of the 5-HT(ID) receptor in the pathophysiology of OCD , by using a challenge paradigm with the selective 5-HT(ID) receptor agonist sumatriptan ( DB00669 ) . A randomized , double-blind , placebo-controlled crossover challenge with sumatriptan ( 100 mg PO ) was performed in 15 OCD patients . Neither the obsessive compulsive symptoms nor mood or anxiety symptoms changed significantly following sumatriptan administration as compared to placebo . Sumatriptan did induce a significant increase in plasma growth hormone ( GH ) levels . In the present study , no indication were found for the role of the 5-HT(ID) receptor in the pathophysiology of OCD . It should be noted , however , that sumatriptan does not readily pass the blood-brain barrier . Selective 5-HT(ID) receptors with better brain penetrating properties may shed more light on the role of this 5-HT receptor subtype in OCD . [ Triptans and calcitonin gene-related peptide ( P80511 ) receptor antagonists ] . While triptans , the P28222 /1D agonists , are effective and generally well-tolerated in many patients up to one-third of migraine patients either may not respond well to triptans , may not tolerate their side effects , or may have contraindications that preclude their use . Recurrence , DB00669 -related side effects , and cardiovascular constriction effects are demerits for acute migraine treatment . P80511 receptor antagonists , the so-called gepants , were clearely designed and expected to be better than triptans . P80511 is located in sensory nerve endings around cranial blood vessels . P80511 is a strong dilator of cerebral arteries and intravenous infusion of P80511 cause a migraine attack . DB04869 is the first selective P80511 receptor antagonist of proven efficacy in migraine . DB04869 could only be administered intravenously and never taken beyond Phase II . Telcagepant is orally available and several completed Phase III trials have revealed positive results . In several comparative studies of telcagepant and triptans , telcegepant did not appeared more effective than zolmitriptan or rizatriptan , although it had fewer DB00669 -related adverse events and drug-related adverse enents . A small number of patients taking olcegepant showed marked elevation in liver transaminase levels . It was decided to discontinue development of olcegepant . New P80511 receptor antagonists would be expected for acute migraine treatment . DB09053 treatment ameliorates murine chronic graft-versus-host disease . Chronic graft-versus-host disease ( cGVHD ) is a life-threatening impediment to allogeneic hematopoietic stem cell transplantation , and current therapies do not completely prevent and/or treat cGVHD . P01730 + T cells and B cells mediate cGVHD ; therefore , targeting these populations may inhibit cGVHD pathogenesis . DB09053 is an FDA-approved irreversible inhibitor of Bruton 's tyrosine kinase ( Q06187 ) and P60568 inducible T cell kinase ( Q08881 ) that targets Th2 cells and B cells and produces durable remissions in B cell malignancies with minimal toxicity . Here , we evaluated whether ibrutinib could reverse established cGVHD in 2 complementary murine models , a model interrogating T cell-driven sclerodermatous cGVHD and an alloantibody-driven multiorgan system cGVHD model that induces bronchiolar obliterans ( BO ) . In the T cell-mediated sclerodermatous cGVHD model , ibrutinib treatment delayed progression , improved survival , and ameliorated clinical and pathological manifestations . In the alloantibody-driven cGVHD model , ibrutinib treatment restored pulmonary function and reduced germinal center reactions and tissue immunoglobulin deposition . Animals lacking Q06187 and Q08881 did not develop cGVHD , indicating that these molecules are critical to cGVHD development . Furthermore , ibrutinib treatment reduced activation of T and B cells from patients with active cGVHD . Our data demonstrate that B cells and T cells drive cGVHD and suggest that ibrutinib has potential as a therapeutic agent , warranting consideration for cGVHD clinical trials . A common single nucleotide polymorphism can exacerbate long-QT type 2 syndrome leading to sudden infant death . BACKGROUND : Identification of infants at risk for sudden arrhythmic death remains one of the leading challenges of modern medicine . We present a family in which a common polymorphism ( single nucleotide polymorphism ) inherited from the father , combined with a stop codon mutation inherited from the mother ( both asymptomatic ) , led to 2 cases of sudden infant death . METHODS AND RESULTS : P51787 , Q12809 , Q14524 , P15382 , Q9Y6J6 , CACNA1c , CACNB2b , and P63252 genes were amplified and analyzed by direct sequencing . Functional electrophysiological studies were performed with the single nucleotide polymorphism and mutation expressed singly and in combination in Chinese ovary ( CHO- P04264 ) and COS-1 cells . An asymptomatic woman presenting after the death of her 2-day-old infant and spontaneous abortion of a second baby in the first trimester was referred for genetic analysis . The newborn infant had nearly incessant ventricular tachycardia while in utero and a prolonged QTc ( 560 ms ) . The mother was asymptomatic but displayed a prolonged QTc . Genetic screening of the mother revealed a heterozygous nonsense mutation ( P926AfsX14 ) in Q12809 , predicting a stop codon . The father was asymptomatic with a normal QTc but had a heterozygous polymorphism ( K897T ) in Q12809 . The baby who died at 2 days of age and the aborted fetus inherited both K897T and P926AfsX14 . Heterologous coexpression of K897T and P926AfsX14 led to loss of function of Q12809 current much greater than expression of K897T or P926AfsX14 alone . CONCLUSIONS : Our data suggest that a common polymorphism ( K897T ) can markedly accentuate the loss of function of mildly defective Q12809 channels , leading to long-QT syndrome-mediated arrhythmias and sudden infant death . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Rizatriptan in migraineurs with unilateral cranial autonomic symptoms : a double-blind trial . The objective and background is to confirm in a double-blind , placebo-controlled study the high DB00669 response rates we had previously reported in an open study in migraine patients with unilateral cranial autonomic symptoms . In this randomized , double-blind , placebo-controlled study 80 migraineurs with unilateral cranial autonomic symptoms were assigned to receive rizatriptan 10 mg wafer or placebo ( ratio 1:1 ) and treated for a single moderate or severe migraine attack . The primary endpoints were pain freedom at 2 h and total migraine freedom at 2 h . Secondary endpoints included pain relief , no associated symptoms and sustained pain freedom or relief . Significantly more patients reported pain freedom at 2 h after taking rizatriptan ( 54 % ) than after placebo ( 8 % ) ( therapeutic gain 46 % [ 28 % ; 64 % ] ; P < 0.001 ) . Similarly , significantly more patients reported total migraine freedom at 2 h after rizatriptan ( 51 % ) than after placebo ( 8 % ) ( therapeutic gain 43 % [ 26 % ; 61 % ] ; P < 0.001 ) . Rizatriptan was also more effective than placebo on most secondary endpoints . We confirm in a placebo-controlled study our previous data suggesting that the presence of unilateral cranial autonomic symptoms in migraineurs predicts a positive response to triptans , probably owing to intense trigeminal peripheral afferent activation which strongly recruits peripheral neurovascular P28222 /1D receptors . Acute and preventive pharmacological trials in migraine should focus also on this subset of migraine patients . A combination of molecular cytogenetic analyses reveals complex genetic alterations in conventional renal cell carcinoma . Here we report the complex pattern of genomic imbalances and rearrangements in a panel of 19 renal cell carcinoma cell lines detected with molecular cytogenetic analysis . Consistent heterogeneity in chromosome number was found , and most cell lines showed a near-triploid chromosome complement . Several cell lines showed deletions of the P04637 ( alias p53 ) , CDKN2A ( alias p16 ) , and P40337 genes . Multiplex fluorescence in situ hybridization ( M- Q5TCZ1 ) analysis revealed chromosome 3 translocated to several other partners chromosomes , as well as breakage events commonly affecting chromosomes 1 , 5 , 8 , 10 , and 17 . The most common abnormality detected with comparative genomic hybridization ( CGH ) was deletions of chromosome 3p , with loss of the Q9NS23 , P49789 , and p44S10 loci frequently involved . CGH gain of 5q showed overrepresentation of the P18146 and P07333 genes . Recurrent alterations to chromosome 7 included rearrangement of 7q11 and gains of the P00533 , O15164 , and P35250 genes . Several lines exhibited rearrangement of 12q11 approximately q14 and overrepresentation of P11802 and SAS loci . M- Q5TCZ1 revealed several other recurrent translocations , and CGH findings included loss of 9p , 14q , and 18q and gain of 8q , 12 , and 20 . Further genomic microarray changes included loss of Q13126 , IGH@ , P28222 , and Q13485 ( previously Q13485 ) and gains of MYC and P11387 . An excellent correlation was observed between the genomic array and Q5TCZ1 data , demonstrating that this technique is effective and accurate . The aberrations detected here may reflect important pathways in renal cancer pathogenesis . UV-induced but P04637 independent apoptotic death in CHO. P04264 cells is promoted by M phase inhibitors . Colocalization of P80511 with P28222 /1D receptors and DB05875 in trigeminal ganglion neurons in rats . Vasodilatation in the dura mater has been implicated in migraine pathogenesis . Anti-migraine DB00669 drugs block vasodilatation by binding to P28222 /1D receptors localized on the peripheral sensory terminals and dural blood vessel smooth muscles . Previous studies suggest that calcitonin gene-related peptide ( P80511 ) released from Adelta-fibres plays a more important role than DB05875 ( SP ) released from C-fibres in inducing dural vasodilatation and that one of the antimigraine mechanisms of DB00669 drugs is inhibiting P80511 release . In the present study , the relationship between P80511 and P28222 /1D receptors , and between P80511 and SP in the trigeminal ganglion neurons in rats was examined by double immunohistochemical staining . P80511 , P28222 , P28221 and SP-positive trigeminal ganglion neurons were all predominantly small and medium-sized . In the trigeminal ganglia , approximately 50 % of P80511 -positive neurons were P28222 positive . Similarly , approximately 55 % of P80511 -positive neurons were P28221 immunoreactive . Approximately 50 % of P80511 -positive neurons were SP-positive , while 93 % of SP-positive neurons were P80511 -positive , suggesting that nearly all SP-positive neurons also contain P80511 . The fibre types of the P28222 - and P28221 -positive neurons were further investigated with an antibody against the A-fibre marker 200-kDa neurofilaments ( NF200 ) . Approximately 46 % of the P28222 -positive and 43 % of the P28221 -positive trigeminal ganglion neurons were also NF200 positive , indicating that many A-fibre trigeminal neurons express P28222 or P28221 receptors . These results support the hypothesis that one important action of antimigraine drugs is the inhibition of P80511 release and that Adelta-fibres may play an important role in migraine pathogenesis . Pharmacokinetic evaluation of frovatriptan . INTRODUCTION : Migraine is the most common painful neurological disorder , affecting 13 % of the general population . Triptans represent a powerful pharmacological tool in acute migraine treatment , however , a significant portion of treated patients can not have access to this class due to possible adverse affects . Today , a total of seven DB00669 molecules are available , representing a commonly prescribed migraine treatment . Although there is a need of extensive use of triptans , only 25 % of migraine patients are using triptans . AREAS COVERED : This review includes triptans and evidence for the use of frovatriptan . A systematic approach is used to discuss the pharmacodynamic and pharmacokinetic aspects of frovatriptan , considering the emerging data on the clinical efficacy of frovatriptan in the treatment of migraine and cluster headaches . The data were obtained by searching the following key words in MEDLINE : pharmacokinetic , pharmacodynamic , triptans , frovatriptan , migraine , menstrual migraine , relatively to the period 1988 - 2011 . EXPERT OPINION : DB00998 has been developed in order to improve safety and efficacy of triptans . It shows a favorable tolerability and efficacy profile , limited to 24/48-h headache recurrence , when compared with other triptans . Preclinical data suggest that the pharmacokinetic profile of frovatriptan may differ from other available triptans . In fact , among triptans , frovatriptan showed the highest potency at the P28222 receptor ( 8.2 ) and the longer half-life ( 26 h ) . These parameters determine the clinical properties of frovatriptan ; in particular the lowest rate of headache recurrence in comparison with other triptans . Migraine : current concepts and emerging therapies . Migraine is a recurrent incapacitating neurovascular disorder characterized by attacks of debilitating pain associated with photophobia , phonophobia , nausea and vomiting . Migraine affects a substantial fraction of world population and is a major cause of disability in the work place . Though the pathophysiology of migraine is still unclear three major theories proposed with regard to the mechanisms of migraine are vascular ( due to cerebral vasodilatation ) , neurological ( abnormal neurological firing which causes the spreading depression and migraine ) and neurogenic dural inflammation ( release of inflammatory neuropeptides ) . The modern understanding of the pathogenesis of migraine is based on the concept that it is a neurovascular disorder . The drugs used in the treatment of migraine either abolish the acute migraine headache or aim its prevention . The last decade has witnessed the advent of Sumatriptan and the ' DB00669 ' class of P28222 /1D receptor agonists which have well established efficacy in treating migraine . Currently prophylactic treatments for migraine include calcium channel blockers , 5-HT2 receptor antagonists , beta adrenoceptor blockers and gamma-amino butyric acid ( GABA ) agonists . Unfortunately , many of these treatments are non specific and not always effective . Despite such progress , in view of the complexity of the etiology of migraine , it still remains undiagnosed and available therapies are underused . In this article , the diverse pieces of evidence that have linked the different theories of migraine with its pathophysiology are reviewed . Furthermore , the present therapeutic targets and futuristic approaches for the acute and prophylactic treatment of migraine , with a special emphasis to calcitonin gene-related peptide , are critically evaluated . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Polyamines and membrane transporters . In recent years , our understanding of the importance of membrane transporters ( MTs ) in the disposition of and response to drugs has increased significantly . MTs are proteins that regulate the transport of endogenous molecules and xenobiotics across the cell membrane . In mammals , two super-families have been identified : DB00171 -binding cassette ( DB01048 ) and solute carrier ( O00585 ) transporters . There is evidence that MTs might mediate polyamines ( PA ) transport . PA are ubiquitous polycations which are found in all living cells . In mammalian cells , three major PA are synthesised : putrescine , spermidine and spermine ; whilst the decarboxylated arginine ( agmatine ) is not produced by mammals but is synthesised by plants and bacteria . In addition , research in the PA field suggests that PA are transported into cells via a specific transporter , the polyamine transport system(s) ( Q03393 ) . Although the Q03393 has not been fully defined , there is evidence that some of the known MTs might be involved in PA transport . In this mini review , eight O00585 transporters will be reviewed and their potential to mediate PA transport in human cells discussed . These transporters are O15245 , O15244 , O75751 , Q96FL8 , P30825 , P08195 , SLC12A8A , and Q86VW1 . Preliminary data from our laboratory have revealed that O15245 might be involved in the PA uptake ; in addition to one member of ABC superfamily ( P08183 protein ) might also mediate the efflux of polyamine like molecules . [ Treatments of migraine ] . During the 1980s , a new class of drugs for the acute treatment of migraine attacks was developed , the triptans . These agents are selective P28222 /1D serotonin receptor agonists , and were developed in order to address the underlying biological mechanism of the migraine attack . French guidelines in migraine are available since 2002 . It is recommended to use a stratified treatment approach during the first consultation with the use of NSAID in first line acute treatment and DB00669 in second line . It is also recommended to use prophylactic treatment when the patient experience frequent and/or severe migraine attack with a bad quality of life and a real impairment . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . The biology of serotonin receptors : focus on migraine pathophysiology and treatment . Serotonin receptors are highly heterogeneous and they have been regrouped within seven different families ( 5-HT1- P34969 ) . With the exception of the 5- Q9H205 which is a ligand-gated ion channel , all others are G-protein coupled receptors with each family sharing structural , pharmacological and transductional characteristics . 5-HT receptors have been implicated in the regulation of several psychiatric and neurological disorders related to serotonergic neurotransmission , and specific receptor subtypes have recently been associated with either the pathogenesis or the treatment of migraine headache . In this respect , activation of vascular P41595 and/or P34969 receptors , possibly as a consequence of the sudden rise in 5-HT levels reported at the onset of a migraine attack , would hypothetically result in dilation of cerebral blood vessels and concomitant activation of sensory trigeminovascular afferents , hence initiating the manifestation of head pain . At this stage in the migraine process , activation of specific subtypes of 5-HT1 receptors has proven clinically effective in relieving migraine pain . Neural P28221 and/or P30939 receptors localized pre-junctionally on trigeminovascular afferents appear to mediate the DB00669 -induced inhibition of the neurogenic inflammatory response , with possible additional sites of action for brain penetrant 5-HT1 receptor agonists in inhibiting the transmission of pain centrally . In contrast , activation of vascular P28222 receptors would constrict meningeal vessels hence recovering their pre-migraine diameter . The recent availability of subtype selective P28221 and P30939 receptor agonists should allow a further test of the neural/vascular hypothesis and could possibly lead to antimigraine drugs with a safer cardiovascular profile . No contractile effect for P28221 and P30939 receptor agonists in human and bovine cerebral arteries : similarity with human coronary artery . 1. Using subtype-selective 5-HT1 receptor agonists and/or the 5-HT1 receptor antagonist GR127935 , we characterized in vitro the 5-HT receptor that mediates the contraction of human and bovine cerebral arteries . Further , we investigated which sumatriptan-sensitive receptors are present in human coronary artery by reverse-transcriptase polymerase chain reaction ( RT-PCR ) . 2 . Agonists with affinity at the P28222 receptor , such as sumatriptan , alniditan and/or IS-159 , elicited dose-dependent contraction in both human and bovine cerebral arteries . They behaved as full agonists at the sumatriptan-sensitive 5-HT1 receptors in both species . In contrast , PNU-109291 and LY344864 , selective agonists at P28221 and P30939 receptors , respectively , were devoid of any significant vasocontractile activity in cerebral arteries , or did not affect the sumatriptan-induced vasocontraction . The rank order of agonist potency was similar in both species and could be summarized as 5-HT = alniditan > sumatriptan = IS-159 >>> PNU-109291 = LY344864 . 3 . In bovine cerebral arteries , the 5-HT1 receptor antagonist GR127935 dose-dependently inhibited the vasoconstrictions elicited by both 5-HT and sumatriptan , with respective pA2 values of 8.0 and 8.6 . 4 . RT-PCR studies in human coronary arteries showed a strong signal for the P28222 receptor while message for the P30939 receptor was weak and less frequently detected . Expression of P28221 receptor mRNA was not detected in any sample . 5 . The present results demonstrate that the DB00669 -induced contraction in brain vessels is mediated exclusively by the P28222 receptor , which is also present in a majority of human coronary arteries . These results suggest that selective P28221 and P30939 receptor agonists might represent new antimigraine drugs devoid of cerebro- and cardiovascular effects . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . [ DB09053 : A new drug of B-cell malignancies ] . DB09053 ( Imbruvica® ) is a first-in-class , orally administered once-daily , that inhibits B-cell antigen receptor signaling downstream of Bruton 's tyrosine kinase ( Q06187 ) . DB09053 has been approved in USA in February 2014 and in France in October 2014 for the treatment of patients with relapsed/refractory mantle cell lymphoma ( Q8WXI8 ) or chronic lymphocytic leukaemia ( CLL ) and for the treatment of patients with CLL and a chromosome 17 deletion ( del 17p ) or P04637 mutation . In clinical studies , ibrutinib induced an impressive overall response rate ( 68 % ) in patients with relapsed/refractory Q8WXI8 ( phase II study ) . In CLL , ibrutinib has shown to significantly improve progression-free survival , response rate and overall survival in patients with relapsed/refractory CLL , including in those with del 17p . DB09053 had an acceptable tolerability profile . Less than 10 % of patients discontinued their treatment because of adverse events . Results are pending in other B-cell lymphomas subtypes such as in diffuse large B-cell lymphoma and in follicular lymphoma . An approval extension has already been enregistered for Waldenström disease in USA in January 2015 . Given its efficacy and tolerability , ibrutinib is an emerging treatment option for patients with B-cell malignancies . [ Rizatriptan : experience after 15 years of clinical use ] . We review the development of rizatriptan , one of the seven P28222 /1D agonists available for the symptomatic treatment of migraine , emphasizing the most relevant contributions carried out from our country . Rizatriptan has shown the quickest onset of action , both in controlled studies and in the different metaanalyses , which translates in high efficacy levels at two hours . Its tolerability and safety profile is similar to that of the other compounds in this pharmacological group . Postlaunching studies have shown that its high efficacy leads to pharmacoeconomic savings and to a robust preference and satisfaction by the patient for this DB00669 . Its efficacy is improved with an early use within migraine attacks and recent data have shown also efficacy in adolescents . This global profile places rizatriptan as a DB00669 of first choice for any kind of migraine attacks . Tissue injury regulates serotonin 1D receptor expression : implications for the control of migraine and inflammatory pain . The anti-migraine action of " DB00669 " drugs involves the activation of serotonin subtype 1D ( P28221 ) receptors expressed on " pain-responsive " trigeminal primary afferents . In the central terminals of these nociceptors , the receptor is concentrated on peptidergic dense core vesicles ( DCVs ) and is notably absent from the plasma membrane . Based on this arrangement , we hypothesized that in the resting state the receptor is not available for binding by a DB00669 , but that noxious stimulation of these afferents could trigger vesicular release of DCVs , thus externalizing the receptor . Here we report that within 5 min of an acute mechanical stimulus to the hindpaw of the rat , there is a significant increase of P28221 -immunoreactivity ( IR ) in the ipsilateral dorsal horn of the spinal cord . We suggest that these rapid immunohistochemical changes reflect redistribution of sequestered receptor to the plasma membrane , where it is more readily detected . We also observed divergent changes in P28221 -IR in inflammatory and nerve-injury models of persistent pain , occurring at least in part through the regulation of P28221 -receptor gene expression . Finally , we found that P28221 -IR is unchanged in the spinal cord dorsal horn of mice with a deletion of the gene encoding the neuropeptide DB05875 . This result differs from that reported for the partial differential-opioid receptor , which is also sorted to DCVs , but is greatly reduced in preprotachykinin mutant mice . We suggest that a " pain " -triggered regulation of P28221 -receptor expression underlies the effectiveness of triptans for the treatment of migraine . Moreover , the widespread expression of P28221 receptor in somatic nociceptive afferents suggests that triptans could , in certain circumstances , treat pain in nontrigeminal regions of the body . DB00184 consumption is regulated by a human polymorphism in dopamine neurons . Smoking is the most important preventable cause of morbidity and mortality worldwide . Recent genome-wide association studies highlighted a human haplotype on chromosome 15 underlying the risk for tobacco dependence and lung cancer . Several polymorphisms in the P32297 - P30532 - P30926 cluster coding for the nicotinic acetylcholine receptor ( nAChR ) α3 , α5 and β4 subunits were implicated . In mouse models , we define a key role in the control of sensitivity to nicotine for the α5 subunit in dopaminergic ( DAergic ) neurons of the ventral tegmental area ( VTA ) . We first investigated the reinforcing effects of nicotine in drug-naive α5(-/-) mice using an acute intravenous nicotine self-administration task and ex vivo and in vivo electrophysiological recordings of nicotine-elicited DA cell activation . We designed lentiviral re-expression vectors to achieve targeted re-expression of wild-type or mutant α5 in the VTA , in general , or in DA neurons exclusively . Our results establish a crucial role for α5*-nAChRs in DAergic neurons . These receptors are key regulators that determine the minimum nicotine dose necessary for DA cell activation and thus nicotine reinforcement . Finally , we demonstrate that a single-nucleotide polymorphism , the non-synonymous α5 variant rs16969968 , frequent in many human populations , exhibits a partial loss of function of the protein in vivo . This leads to increased nicotine consumption in the self-administration paradigm . We thus define a critical link between a human predisposition marker , its expression in DA neurons and nicotine intake . Serotonergic regulation of somatosensory cortical development : lessons from genetic mouse models . Monoaminergic neurotransmitter systems appear early during embryogenesis , suggesting that they could play important roles in brain development . Accumulated evidence indicates that serotonin ( 5-hydroxytryptamine , 5-HT ) regulates neural as well as nonneural development , including early aspects of embryonic development , differentiation of neuronal progenitors , and morphogenesis of the craniofacial region , heart and limb . Recent studies using monoamine oxidase-A ( P21397 ) , 5-HT transporter , vesicular monoamine transporter-2 ( Q05940 ) and P28222 receptor single , double and triple knockout mice have provided evidence that the serotonergic system plays important roles in barrel field formation in the developing somatosensory cortex . Here we review evidence from these genetic mouse models and , based on the accumulated evidence , propose a testable model for future studies of mechanisms underlying serotonergic regulation of cortical development . Emerging therapeutic options for acute migraine : focus on the potential of lasmiditan . The serotonin receptor agonist DB00669 drugs ( P28222 /1D receptor agonists ) have been in use for over 20 years in the abortive management of migraine . Although clearly effective , their ability to produce vasoconstriction in cerebral and coronary arteries , thought to be mediated by their high affinity for the P28222 receptor , has been a limitation to their use in certain patient populations . Variable potency DB00669 binding at the P30939 receptor occurs in addition to binding at the P28222 and P28221 receptors . A more selective serotonin agonist without P28222 -mediated vasoconstriction might prove efficacious yet safer . The P30939 receptor has been targeted as a site of action for such a drug . In experimental models , P30939 receptor agonists have been shown to block neurogenic inflammation and c-Fos expression in neural tissue and , as well , show no evidence of vasoconstriction in vascular tissue models in vitro . In clinical trials , efficacy in the abortive management of migraine has been established . Lasmiditan ( LY573144 ) , a selective P30939 receptor agonist ( P04264 =2.21 μM ) , showed efficacy in its primary endpoint , with a 2-hour placebo-subtracted headache response of 28.8 % , though with frequent reports of dizziness , paresthesias , and vertigo . Study results support an emerging central neuronal mechanism of migraine pathophysiology . This review traces the history and use of P30939 receptor agonists , now referred to as neurally acting anti-migraine agents in migraine management . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . The efficacy and tolerability of frovatriptan and dexketoprofen for the treatment of acute migraine attacks . DB00998 is a DB00669 characterized by a high affinity for P28222 /1D receptors and a long half-life contributing to a more sustained and prolonged action than other triptans . DB09214 is a nonsteroidal anti-inflammatory drug with a relatively short half-life and rapid onset of action , blocking the action of cyclo-oxygenase , which is involved in prostaglandins ' production , thus reducing inflammation and pain . Both drugs have been successfully employed as monotherapies for the treatment of acute migraine attacks . The combination of these two drugs ( frovatriptan 2.5 mg plus dexketoprofen 25 or 37.5 mg ) has been tested in migraine sufferers , showing a rapid and good initial efficacy , with 2-h pain free rates of 51 % , and a high persistence in the 48-h following the onset of pain : recurrence occurred in only 29 % of attacks and sustained pain free rates were 43 % at 24- and 33 % at 48-h . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients .

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Genome-wide association study identifies genetic determinants of warfarin responsiveness for Japanese . DB00682 is a commonly used anticoagulant , whose dose needs to be determined for each individual patient owing to large inter-individual variability in its therapeutic dose . Although several clinical and genetic variables influencing warfarin dose have been identified , uncovering additional factors are critically important for safer use of warfarin . Through a genome-wide association study , we identified single-nucleotide polymorphism ( SNP ) rs2108622 [ cytochrome P450 , family 4 , subfamily F , polypeptide 2 ( P78329 ) ] as a genetic determinant of warfarin responsiveness for Japanese . Stratifying subjects who have been pre-classified according to the genotypes of SNP rs10509680 [ cytochrome P450 , family 2 , subfamily C , polypeptide 9 ( P11712 ) ] and SNP rs9923231 [ vitamin K epoxide reductase complex subunit 1 ( Q9BQB6 ) ] , based on their genotypes of rs2108622 allowed identification of subjects who require higher dose of warfarin . Incorporating genotypes of rs2108622 into a warfarin dosing algorithm that considers age , body surface area , status of amiodarone co-administration and genotypes of SNPs in the P11712 and Q9BQB6 genes improved the model 's predictability to 43.4 % . In this study , the association of P78329 with warfarin dose of the Japanese has been established for the first time . Besides , a warfarin dosing algorithm that incorporates genotypes of rs2108622 and amiodarone co-administration status was suggested for the Japanese . Our study also implied that common SNPs other than those in the P11712 , Q9BQB6 and P78329 genes that show strong effect on the therapeutic warfarin dose might not exist . Exploring the roles of P22309 and P35503 in oral clearance of GSK2190915 , a P09917 -activating protein inhibitor . Pharmacokinetic variability in drug exposure is a concern for all compounds in development including those for the treatment of asthma and other respiratory disorders . Substantial variability in the oral clearance of GSK2190915 , a P09917 -activating protein inhibitor that attenuates the production of leukotriene B4 and cysteinyl leukotrienes , is largely unaccounted for by clinical variables . A study of 41 patients , 78 % ( 32/41 ) of whom were non-Hispanic whites , with mild to moderate asthma identified an association of P22309 *28 and P35503 *2 with the oral clearance of GSK2190915 ( P=3.8×10⁻⁴ and 1.2×10⁻⁵ , respectively ) . However , in a subsequent replication study of 403 non-Hispanic white patients with asthma , we failed to observe a statistically significant association between oral clearance of GSK2190915 and either P22309 *28 or P35503 *2 ( P > 0.05 ) . Therefore , genetic effects that could explain the systemic exposure level variability of GSK2190915 were not identified . Effects of DB00482 and DB00744 on liver metastasis and lipidperoxidation in pancreatic cancer in Syrian hamsters . Selective inhibition of eicosanoid synthesis is thought to have effects on carcinogenesis in lung and colon cancer . However , it is still unknown whether pancreatic cancer might also be influenced . Therefore we evaluated the impact of selective cyclooxygenase-2 inhibitor DB00482 and selective P09917 inhibitor DB00744 on liver metastasis in a solid model of pancreatic adenocarcinoma in Syrian hamster . In week 33 , the animals were sacrificed and incidence of pancreatic carcinomas and number and size of liver metastases were determined . Activities of antioxidative enzymes ( GSHPX/SOD ) and concentrations of products of lipidperoxidation were measured in liver metastases and non-metastatic hepatic tissue . The incidence ( 54.5 vs. 100 % ) , number ( 3.17 +/- 0.98 vs. 6.75 +/- 0.71 ) and size ( 2.67 +/- 1.97 vs. 11.75 +/- 1.98 mm2 ) of liver metastases were decreased by combined therapy of DB00744 and DB00482 ( P < 0.05 ) . Furthermore , activities of GSHPX ( [ 73.77 +/- 5.67 ] *10(5) vs. [ 15.49 +/- 4.02 ] *10(5) U/mg prot. ; P < 0.05 ) and SOD ( 474.92 +/- 108.8 vs. 127.89 +/- 38.75 U/mg prot. ; P < 0.05 ) were increased , while lipidperoxidation ( 0.31 +/- 0.08 nmol/mg prot. vs. 1.54 +/- 0.55 nmol/mg prot. ; P < 0.05 ) was decreased by combination therapy , in non-metastatic hepatic tissue . Moreover , combined therapy increased lipidperoxidation in liver metastases ( 0.47 +/- 0.09 vs. 1.95 +/- 0.12 nmol/mg prot. ; P < 0.05 ) . Thus , a combination of DB00482 and DB00744 might be a new concept to decrease tumour growth in liver metastases in advanced pancreatic cancer . Preliminary assessment of differential expression of candidate genes associated with atherosclerosis . Identifying susceptible genes associated with the pathogenesis of atherosclerosis ( ATH ) may contribute toward better management of this condition . This preliminary study was aimed at assessing the expression levels of 11 candidate genes , namely tumor protein ( P04637 ) , transforming growth factor , beta receptor II ( P37173 ) , cysthathionenine-beta-synthase ( P35520 ) , insulin receptor substrate 1 ( P35568 ) , lipoprotein lipase ( P06858 ) , methylenetetrahydrofolate reductase ( P42898 ) , thrombomodulin ( P07204 ) , lecithin-cholesterol acyltransferase ( P04180 ) , matrix metallopeptidase 9 ( P14780 ) , low density lipoprotein receptor ( P01130 ) , and arachidonate P09917 -activating protein ( P20292 ) genes associated with ATH . Twelve human coronary artery tissues ( HCATs ) were obtained from deceased subjects who underwent post-mortem procedures . Six atherosclerotic coronary artery tissue ( ACAT ) samples representing the cases and non-atherosclerotic coronary artery tissue ( NCAT ) samples as controls were gathered based on predetermined inclusion and exclusion criteria . Gene expression levels were assessed using the GenomeLab Genetic Analysis System ( GeXP ) . The results showed that P01130 , P04637 , and P14780 expression levels were significantly increased in ACAT compared to NCAT samples ( p < 0.05 ) . Thus , P01130 , P04637 , and P14780 genes may play important roles in the development of ATH in a Malaysian study population . 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 . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Quantum mechanics-based properties for 3D-QSAR . We have used a set of four local properties based on semiempirical molecular orbital calculations ( electron density ( ρ ) , hydrogen bond donor field ( HDF ) , hydrogen bond acceptor field ( P00748 ) , and molecular lipophilicity potential ( MLP ) ) for 3D-QSAR studies to overcome the limitations of the current force field-based molecular interaction fields ( MIFs ) . These properties can be calculated rapidly and are thus amenable to high-throughput industrial applications . Their statistical performance was compared with that of conventional 3D-QSAR approaches using nine data sets ( angiotensin converting enzyme inhibitors ( P12821 ) , acetylcholinesterase inhibitors ( AchE ) , benzodiazepine receptor ligands ( BZR ) , cyclooxygenase-2 inhibitors ( P35354 ) , dihydrofolate reductase inhibitors ( P00374 ) , glycogen phosphorylase b inhibitors ( GPB ) , thermolysin inhibitors ( THER ) , thrombin inhibitors ( THR ) , and serine protease factor Xa inhibitors ( fXa ) ) . The 3D-QSAR models generated were tested thoroughly for robustness and predictive ability . The average performance of the quantum mechanical molecular interaction field ( QM-MIF ) models for the nine data sets is better than that of the conventional force field-based MIFs . In the individual data sets , the QM-MIF models always perform better than , or as well as , the conventional approaches . It is particularly encouraging that the relative performance of the QM-MIF models improves in the external validation . In addition , the models generated showed statistical stability with respect to model building procedure variations such as grid spacing size and grid orientation . QM-MIF contour maps reproduce the features important for ligand binding for the example data set ( factor Xa inhibitors ) , demonstrating the intuitive chemical interpretability of QM-MIFs . Enhanced fracture repair by leukotriene antagonism is characterized by increased chondrocyte proliferation and early bone formation : a novel role of the cysteinyl LT-1 receptor . Inflammatory mediators and drugs which affect inflammation can influence the healing of injured tissues . Leukotrienes are potent inflammatory mediators , and similar to prostaglandins , are metabolites of arachidonic acid which can have positive or negative effects on bone and cartilage tissues . Here we tested the hypothesis that blocking the negative regulation of leukotrienes , would lead to enhanced endochondral bone formation during fracture repair . A closed femoral fracture was created in mice . Animals were divided into three groups for treatment with either montelukast sodium , a cysteinyl leukotriene type 1 receptor antagonist ( trade name Singulair ) , zileuton , a P09917 enzyme inhibitor ( trade name DB00744 ) , or carrier alone . The fractures were analyzed using radiographs , quantitative gene expression , histology and histomorphometry , and immunohistochemistry . Both the montelukast sodium group and the zileuton group exhibited enhanced fracture repair when compared with controls . Both treatment groups exhibited increased callous size and earlier bone formation when compared to controls as early as day 7 . Gene expression analysis of treatment groups showed increased markers of chondrocyte proliferation and differentiation , and increased early bone formation markers when compared with controls . Treatment with montelukast sodium directly targeted the cysteinyl leukotriene type 1 receptor , leading to increased chondrocyte proliferation at early time points . These novel findings suggests a potential mechanism by which the cysteinyl leukotriene type 1 receptor acts as a negative regulator of chondrocyte proliferation , with important and previously unrecognized implications for both fracture repair , and in a broader context , systemic chondrocyte growth and differentiation . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . DB04725 inhibits interleukin-18-induced pro-inflammatory cytokine release and cellular proliferation in human mesangial cells . DB04725 , a novel dual anti-inflammatory drug that inhibits P09917 ( 5- P28300 ) and cyclooxygenase ( P36551 ) , has recently been defined to have therapeutic effects in osteoarthritis . Both 5- P28300 and P36551 play functional roles in the pathogenesis of glomerulonephritis in children as well . Q14116 is a pro-inflammatory cytokine . It remains unclear whether licofelone can ameliorate inflammatory response of human mesangial cells ( HMC ) exposed to interleukin-18 . In this study , HMC were cultured and exposed to interleukin-18 with or without pre-treatment of licofelone . P35354 and 5- P28300 enzyme activities in mesangial cells were determined with chromometry or high-performance liquid chromatography . DB00917 , cysteinyl leukotriene , monocyte chemotactic protein-1 and interferon-γ concentrations in culture medium were measured using an enzyme-linked immunosorbent assay . Western blotting was employed to detect phosphorylated mitogen-activated protein kinases P27361 /2 , p38 and P45983 /2 in HMC . It was found that licofelone attenuated interleukin-18-induced P35354 enzyme activity in HMC and prostaglandin E2 release in a dose-dependent manner . Similarly , licofelone inhibited interleukin-18-induced 5- P28300 enzyme activity and leukotriene release . DB04725 reduced interleukin-18-induced phosphorylation of p38 mitogen-activated protein kinase and suppressed monocyte chemotactic protein-1 and interferon-γ synthesis . Moreover , licofelone inhibited Q14116 -induced proliferation of mesangial cells . We conclude that licofelone inhibits interleukin-18-induced pro-inflammatory cytokine release and cellular proliferation in HMC , which may represent a really interesting therapeutic approach for glomerulonephritis in children . Inhibition of a thrombin anion-binding exosite-2 mutant by the glycosaminoglycan-dependent serpins protein C inhibitor and heparin cofactor II . Antithrombin ( P01008 ) , heparin cofactor II ( HCII ) and protein C inhibitor ( P05154 ; also named plasminogen activator inhibitor-3 ) are serine protease inhibitors ( serpins ) whose thrombin inhibition activity is accelerated in the presence of glycosaminoglycans . We compared the inhibition properties of P05154 and HCII to P01008 using R93A/R97A/R101A thrombin , an anion-binding exosite-2 ( exosite-2 ) mutant that has greatly reduced heparin-binding properties . DB01109 -enhanced P05154 inhibition of R93A/R97A/R101A thrombin was only approximately 2-fold compared to 40-fold enhancement with wild-type recombinant thrombin . P07204 ( TM ) ( with or without the chondroitin sulfate moiety ) accelerated P05154 inhibition of both wild-type and R93A/R97A/R101A thrombins . HCII achieved the same maximum activity in the presence of heparin with both wild-type and R93A/R97A/R101A thrombins ; however , the optimum heparin concentration was 20 times greater than the reaction with wild-type thrombin , indicative of a decrease in heparin affinity . Dermatan sulfate ( DSO4 ) -catalyzed HCII thrombin inhibition was unchanged in R93A/R97A/R101A thrombin compared to wild-type recombinant thrombin . These results suggest that P05154 is similar to P01008 and depends upon ternary complex formation with heparin and these specific thrombin exosite-2 residues to accelerate thrombin inhibition . In contrast , HCII does not require DB00125 (93) , DB00125 (97) and DB00125 (101) of thrombin exosite-2 and further supports the hypothesis that HCII uses an allosteric process following glycosaminoglycan binding to inhibit thrombin . DB01016 -induced apoptosis is specifically enhanced by expression of the sulfonylurea receptor isoform Q09428 but not by expression of SUR2B or the mutant Q09428 (M1289T) . Q09428 ( Q09428 ) is the regulatory subunit of the pancreatic DB00171 -sensitive K+ channel ( K( DB00171 ) channel ) , which is essential for triggering insulin secretion via membrane depolarization . Sulfonylureas , such as glibenclamide and tolbutamide , act as K( DB00171 ) channel blockers and are widely used in diabetes treatment . These antidiabetic substances are known to induce apoptosis in pancreatic beta-cells or beta-cell lines under certain conditions . However , the precise molecular mechanisms of this sulfonylurea-induced apoptosis are still unidentified . To investigate the role of Q09428 in apoptosis induction , we tested the effect of glibenclamide on recombinant human embryonic kidney 293 cells expressing either Q09428 , the smooth muscular isoform SUR2B , or the mutant Q09428 (M1289T) at which a single amino acid in transmembrane helix 17 ( TM17 ) was exchanged by the corresponding amino acid of SUR2 . By analyzing cell detachment , nuclear condensation , DNA fragmentation , and caspase-3-like activity , we observed a Q09428 -specific enhancement of glibenclamide-induced apoptosis that was not seen in SUR2B , Q09428 (M1289T) , or control cells . Coexpression with the pore-forming Kir6.2 subunit did not significantly alter the apoptotic effect of glibenclamide on Q09428 cells . In conclusion , expression of Q09428 , but not of SUR2B or Q09428 (M1289T) , renders cells more susceptible to glibenclamide-induced apoptosis . Therefore , Q09428 as a pancreatic protein could be involved in specific variation of beta-cell mass and might also contribute to the regulation of insulin secretion at this level . According to our results , TM17 is essentially involved in Q09428 -mediated apoptosis . This effect does not require the presence of functional Kir6.2-containing K( DB00171 ) channels , which points to additional , so far unknown functions of Q09428 . Leukotriene B4 amplifies NF-κB activation in mouse macrophages by reducing O15524 inhibition of MyD88 expression . Activation of NF-κB and P09917 -mediated ( P09917 -mediated ) biosynthesis of the lipid mediator leukotriene B4 ( LTB4 ) are pivotal components of host defense and inflammatory responses . However , the role of LTB4 in mediating innate immune responses elicited by specific TLR ligands and cytokines is unknown . Here we have shown that responses dependent on MyD88 ( an adaptor protein that mediates signaling through all of the known TLRs , except O15455 , as well as IL-1β and Q14116 ) are reduced in mice lacking either P09917 or the LTB4 receptor BTL1 , and that macrophages from these mice are impaired in MyD88-dependent activation of NF-κB . This macrophage defect was associated with lower basal and inducible expression of MyD88 and reflected impaired activation of P42224 and overexpression of the P42224 inhibitor O15524 . Expression of MyD88 and responsiveness to the O00206 ligand LPS were decreased by Stat1 siRNA silencing in WT macrophages and restored by Socs1 siRNA in P09917 -deficient macrophages . These results uncover a pivotal role in macrophages for the GPCR Q15722 in regulating activation of NF-κB through Stat1-dependent expression of MyD88 . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . DB00104 -targeted liposomes loaded with CPT-11 enhanced cytotoxicity for the treatment of medullary thyroid carcinoma . Medullary thyroid carcinoma ( P04629 ) is a rare endocrine tumor that frequently metastasizes , but treatment with irinotecan ( CPT-11 ) is limited because of side effects . P04629 is known to overexpress the somatostatin receptor subtype 2 ( P30874 ) . DB00104 ( Oct ) is a somatostatin analogue that has a high binding affinity for SSTR and can be used as a tumor-targeting ligand . We prepared Oct-targeted liposomes loaded with CPT-11 using Oct-poly ( ethylene glycol ) ( PEG ) -lipid and evaluated Oct-mediated association and cytotoxicity of the liposomes with an P04629 cell line TT . The association of higher concentrations of modified Oct-targeted liposomes with TT cells was significantly higher than PEGylated liposomes and was significantly inhibited by empty Oct-targeted liposomes but not by free Oct . With exposure for 96 h , the cytotoxicity of Oct-targeted liposomal CPT-11 ( IC50 : 1.05 ± 0.47 μM ) was higher than free CPT-11 ( IC50 : 3.76 ± 0.61 μM ) or PEGylated liposomal CPT-11 ( IC50 : 3.05 ± 0.28 μM ) . In addition , empty Oct-targeted liposomes showed significantly higher cytotoxicity than empty nontargeted liposomes at a concentration where free Oct did not show cytotoxicity , suggesting that Oct as a ligand showed cytotoxicity . Moreover , Oct-targeted liposomal CPT-11 led to significantly higher antitumor activity and prolonged the survival time compared with nontargeted liposomal and free CPT-11 at a one-third dose and lower administration times with free CPT-11 . These findings indicated that Oct-targeted liposomes loaded with CPT-11 may offer considerable potential for P04629 chemotherapy because cytotoxicity of both CPT-11 and Oct was enhanced by effective cellular uptake via P30874 . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . DB00227 induces the expression of bradykinin type 2 receptors in cultured human coronary artery endothelial cells . Cardioprotective bradykinin type-2 receptors ( BK-2Rs ) are downregulated in the myocardial endothelium of both human and rat failing hearts . Statins are cardioprotective drugs that reduce the level of plasma cholesterol but also exert cholesterol-independent pleiotropic effects . Here we examined the effect of lovastatin on BK-2R expression in cultured human coronary artery endothelial cells . The effect of lovastatin on the expression of BK receptors in human coronary artery endothelial cells ( HCAECs ) was examined by real-time PCR , Western blot analysis and immunocytochemistry . DB00227 induced a time- and concentration-dependent increase in both BK-2R and BK-1R mRNA expression in the cultured HCAECs . Also , the number of functional BK-2Rs capable of inducing BK-mediated NO production and cGMP signaling was increased in the lovastatin-treated HCAECs . Mevalonate , the direct metabolite of P04035 , reversed the effect of lovastatin . Furthermore , lovastatin inhibited Rho activation and a selective inhibitor of Rho-associated kinases , Y-27632 , induced a similar increase in BK-2R expression as lovastatin . In contrast , a specific inhibitor of P35354 , NS398 , significantly inhibited the lovastatin-induced expression of BK-2Rs . Here we show for the first time that lovastatin induces the expression of BK-2Rs in cultured human coronary artery endothelial cells through a novel cholesterol-independent pleiotropic mechanism that involves RhoA kinase inhibition and P35354 activation . Thus , reported beneficial effects of statins in cardiovascular diseases may be partly mediated by an increased expression of cardioprotective BK-2Rs in the endothelial cells of the coronary tree . Moreover , the use of P35354 inhibitors may affect the level of endothelial BK-2Rs in a negative fashion . Effect of cyclooxygenase inhibitors on the P06850 -induced pituitary-adrenocortical activity during crowding stress . The aim of the present study was to determine the effect of social stress and significance of prostaglandins ( PG ) generated by constitutive and inducible cyclooxygenase ( P23219 and P35354 ) in the stimulation of hypothalamic-pituitary-adrenal ( Q9Y251 ) axis by corticotropin releasing hormone ( P06850 ) under basal and social crowding stress conditions . The stressed rats were crowded in groups of 24 to a cage for 3 or 7 days , whereas the control animals were haused in groups of 7 to a cage of the same size . The activity of Q9Y251 axis was determined by measuring plasma DB01285 and serum corticosterone levels 1 h after i.p . P06850 administration . Inhibitors of P23219 , piroxicam ( 0.2 , 2.0 , and 5.0 mg/kg ) , and P35354 , compound NS-398 ( 0.2 and 2.0 mg/kg ) , were administered i.p . 15 min prior to P06850 ( 0.1 microg/kg i.p. ) to control or crowded rats . The obtained results indicate that social stress for 3 and 7 days markedly intensifies the stimulatory action of P06850 on DB01285 secretion . Neither piroxicam nor NS-398 induce any significant effect on the P06850 -elicited DB01285 and corticosterone secretion in non-stressed or crowded rats . Therefore , PG generated by P23219 or P35354 do not participate to a significant extent in the stimulation of Q9Y251 axis by P06850 under either basal conditions or during crowding stress . These results also indicate that the stimulatory action of P06850 on DB01285 secretion is not only completely resistant to desensitization but is sensitized during social crowding stress . The results contrast with a significant involvement of PG in the vasopressin-induced stimulation of Q9Y251 response during crowding stress . Interaction between P20292 and P20815 gene variants significantly increases the risk for cerebral infarctions in Chinese . In this study , we investigated associations between susceptibility genes and cerebral infarctions in a Chinese population , and whether gene-gene interactions increase the risk of cerebral infarctions . Overall , 292 patients with cerebral infarctions and 259 healthy control individuals were included . Eight variants in five candidate genes were examined for the risk of stroke , including the SG13S32 ( rs9551963 ) , SG13S42 ( rs4769060 ) , SG13S89 ( rs4769874 ) , and SG13S114 ( rs10507391 ) variants of the P09917 activating protein ( P20292 ) gene , the G860A ( rs751141 ) variant of the soluble epoxide hydrolase ( P34913 ) gene , the A1075C ( rs1057910 ) variant of the P11712 *2 gene , the C430T ( rs1799853 ) variant of the P11712 *3 gene , and the A6986G ( rs776746 ) variant of the P20815 gene . Gene-gene interactions were explored using generalized multifactor dimensionality reduction methods . There were no statistically significant differences in the frequencies of the genotypes of the eight candidate genes . The generalized multifactor dimensionality reduction analysis showed a significant gene-gene interaction between SG13S114 and A6986G , with scores of 10 for cross-validation consistency and 9 for the sign test ( P=0.0107 ) . These gene-gene interactions predicted a significantly higher risk of cerebral infarction ( adjusted for age , hypertension , and diabetes mellitus ; odds ratio=1.80495 % , confidence interval : 1.180-2.759 , P=0.006 ) . A two-loci gene interaction confers a significantly higher risk for cerebral infarction . The combinational analysis used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases . Cytokine-modulating activity of tepoxalin , a new potential antirheumatic . Tepoxalin is a new dual cyclooxygenase/ P09917 anti-inflammatory compound currently under clinical investigation . It has been shown to possess anti-inflammatory activity in a variety of animal models and more recently to inhibit P60568 induced signal transduction . The current study was conducted to evaluate the cytokine modulating activity of tepoxalin and the role of iron in these effects . In human peripheral blood mononuclear cells ( PBMC ) stimulated with OKT3/PMA , tepoxalin inhibited lymphocyte proliferation with an IC50 of 6 microM . Additionally , it inhibited the production of LTB4 ( IC50 = 0.5 microM ) and the cytokines P60568 , P05231 and P01375 alpha ( IC50 = 10-12 microM ) . Cytotoxicity was not demonstrated at these concentrations . Add-back experiments with either cytokines ( P60568 or P05231 ) , LTB4 or conditioned media failed to restore the proliferative response in the presence of tepoxalin . However , the concurrent addition of iron ( in the form of ferrous or ferric chloride and other iron salts ) reversed the inhibition of proliferation caused by tepoxalin . Tepoxalin also inhibits the activation of NF kappa B , a transcription factor which acts on several cytokine genes . Tepoxalin 's effect on NF kappa B is also reversed by the addition of iron salts . These data suggest that the action of tepoxalin to inhibit proliferation in PBMC may be at least in part due to its ability to reduce the amount of available iron resulting in decreased activation of NF kappa B and subsequent inhibition of cytokine production . Enhancement of 5-fluorouracil efficacy on high P35354 expressing HCA-7 cells by low dose indomethacin and NS-398 but not on low P35354 expressing HT-29 cells . The antiproliferative effect of 5-fluorouracil ( DB00544 ) in the presence of low dose non-steroidal anti-inflammatory drugs ( NSAIDs ) on high cyclooxygenase-2 ( P35354 ) -expressing HCA-7 and low P35354 -expressing HT-29 colon carcinoma cell lines was investigated . Pharmacogenetic parameters were studied to characterize the DB00544 sensitivity of the two cell lines . P04818 ( TS ) and methylenetetrahydrofolate reductase ( P42898 ) polymorphisms were determined by PCR analysis . Cell proliferation was measured by P50991 assay , cell cycle distribution and apoptosis by FACS analysis . Cyclooxygenase expression was detected by Western blot and also by fluorescence microscopy . Prostaglandin E(2) ( PGE(2) ) levels were investigated with ELISA kit . The HT-29 cell line was found to be homozygous for TS 2R and 1494ins6 and T homozygous for P42898 677 polymorphisms predicting high DB00544 sensitivity ( IC(50) : 10 microM ) . TS 3R homozygosity , TS 1496del6 and P42898 677CT heterozygosity may explain the modest DB00544 sensitivity ( IC(50) : 1.1 mM ) of the HCA-7 cell line . Indomethacin and NS-398 ( 10 microM and 1.77 microM , respectively ) reduced the PGE(2) level in HCA-7 cells ( > 90 % ) . Low concentrations of NSAIDs without antiproliferative potency increased the S-phase arrest and enhanced the cytotoxic action of DB00544 only in HCA-7 cells after 48-hours treatment . The presented data suggested that the enhancement of DB00544 cytotoxicity by indomethacin or NS-398 applied in low dose is related to the potency of NSAIDs to modulate the cell-cycle distribution and the apoptosis ; however , it seems that this effect might be dependent on cell phenotype , namely on the P35354 expression . [ Effect of the monophase oral contraceptive combination with 20 ug ethinyl estradiol/150 ug desogestrel on haemostasis ] . The authors examined the changes in the haemostasis during the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel at 35 women , a basic group , who used the oral contraceptive in the duration of 12 months and a control group ( n=35 ) , who do not use the pills . We found statistically significant increase of Antithrombin III ( P01008 ) ( p < 0.011 ) , Cofactor II of DB01109 ( HCII ) ( p < 0.001 ) , the activity of plasminogen ( p < 0.026 ) and beta2-antiplasmin ( 0.026 ) , significant decrease of P02810 ( PrC ) ( p < 0.0001 ) and of total Protein S ( TPrS ) ( p < 0.03 ) in the basic group in comparision with the control one . We do not observe significant changes in the rest of the haemostatic variables between the two groups . During the use of the oral contraceptive combination with 20 microg ethynil estradiol/150 microg desogestrel the changes in the system of the natural inhibitors are balanced by these in the system of fibrinolysis . DB00227 , a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3-Hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization/flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3-dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer . DB00563 in rheumatoid arthritis : studies with animal models . The present studies have shown that low doses of methotrexate can suppress the inflammation and joint destruction associated with animal models of arthritis . The antiinflammatory effects of methotrexate are probably related to its inhibitory effect on chemotaxis . At the low doses used , methotrexate does not induce systemic immunosuppression . In methotrexate-treated rats , an improvement in P60568 synthesis is observed and increases in P60568 levels are expected to improve cell mediated immunity . Suppressor cells appear to be very sensitive to methotrexate . Macrophage function is modulated by methotrexate . All of these effects including the effects on joint destruction are probably due to inhibition of P00374 activity of critical cells that are involved in the pathogenesis of rat arthritis induced either by adjuvant or by streptococcal cell walls . Some of these effects have been extended to human arthritis but additional studies are required to understand how low dose methotrexate exerts its beneficial effects in humans . Suppression of cyclooxygenase-2 overexpression by 15S-hydroxyeicosatrienoic acid in androgen-dependent prostatic adenocarcinoma cells . Emerging reports now implicate alterations of arachidonic acid ( AA ) metabolism with prostate carcinogenesis . To test this hypothesis , androgen-primed benign hyperplastic ( BHC ) and malignant tumorigenic ( P04629 ) cells derived from the Lobund-Wistar rat model of autochthonous prostate adenocarcinoma were incubated with (14)C-AA . Our data using MTCs revealed enhanced dual metabolism of (14)C-AA via P36551 to generate increased PGE(2) and via P09917 ( P28300 ) to generate increased 5S-HETE in tumorigenic cells . Western blot of MTCs revealed upregulation of P35354 expression . This paralleled the increased biosynthesis of PGE(2) . Since some polyunsaturated fatty acids have been reported to modulate AA metabolism and tumorigenesis , we primed the cells with either gamma-linolenic acid ( GLA ) or its in vivo metabolite , 15S-HETrE , prior to incubation with AA . Our data revealed suppression of P35354 expression/PGE(2) biosynthesis . In parallel , priming cells with 15S-HETrE resulted in greater suppression of P35354 expression/PGE(2) biosynthesis . These findings suggest that 15S-HETrE could function in vivo after dietary intake of GLA to suppress DB02901 -enhanced prostatic P35354 expression/PGE(2) biosynthesis and , thus , alleviate tumor growth and progression . Effects of selective P35354 and 5- P28300 inhibition on prostaglandin and leukotriene synthesis in ductal pancreatic cancer in Syrian hamster . Selective inhibition of eicosanoid synthesis seems to decrease carcinogenesis , however , the effect on liver metastasis in pancreatic cancer is still unknown . Ductal pancreatic adenocarcinoma was chemically induced by weekly injection of N-nitrosobis-2-oxopropylamine ( BOP ) in Syrian hamster . Animals received selective inhibition of cyclooxygenase-2 ( DB00482 ) and P09917 ( DB00744 ) . In week 33 , hamsters were sacrificed and incidence of pancreatic carcinomas as well as liver metastases were examined . Furthermore , size and number of liver metastases per animal were determined and concentration of PGF1alpha , DB00917 and leukotrienes was measured in hepatic and pancreatic tissue . Combined therapy ( DB00482 + DB00744 ) significantly decreased incidence , number and size of liver metastases . Furthermore extra- and intrametastatic concentration of DB00917 was reduced by this treatment in hepatic tissue . Single Cox-2-inhibition ( DB00482 ) decreased intrametastatic hepatic PGF1alpha and DB00917 concentration while PGF1alpha concentration was reduced in non-metastatic liver ( nml ) . Moreover 5- P28300 -inhibition ( DB00744 ) decreased intrametastatic DB00917 concentration as well as PGF1alpha and DB00917 in nml . In pancreatic carcinomas highest LT-concentration was found after combined treatment and this therapy group was the only one revealing a significantly higher amount of LTs in carcinomas compared to tumour-free tissue . Hepatic LT-concentration was significantly lower in the control groups than in nml of the tumour groups . Combination of Cox-2-inhibition and 5-Lox-inhibition might be a suitable adjuvant therapy to prevent liver metastasis in human ductal pancreatic adenocarcinoma . Nonsteroidal anti-inflammatory drugs ( NSAID ) sparing effects of glucosamine hydrochloride through N-glycosylation inhibition ; strategy to rescue stomach from NSAID damage . Gastrointestinal or cardiovascular complications limit nonsteroidal anti-inflammatory drugs ( NSAID ) prescription . DB01296 hydrochloride ( GS-HCl ) alternatively chosen , but debates still exist in its clinical efficiency . P35354 instability through inhibiting P35354 N-glycosylation of GS-HCl raised the possibility of NSAID sparing effect . Study was done to determine whether combination treatment of glucosamine and NSAID contributes to gastric safety through NSAID sparing effect . IEC-6 cells were stimulated with P01375 -α and compared the expressions of inflammatory mediators after indomethacin alone or combination of indomethacin and GS-HCl by Western blotting and RT-PCR . C57BL/6 mice injected with type II collagen to induce arthritis were treated with indomethacin alone or combination of reduced dose of indomethacin and GS-HCl after 3 weeks . P01375 -α increased the expression of P35354 , P35228 and inflammatory cytokines , but GS-HCl significantly attenuated P01375 -α-induced P35354 expression . Decreased P35354 after GS-HCl was caused by N-glycosylation inhibition as much as tunicamycin . Combination of reduced dose of indomethacin and GS-HCl significantly reduced the expressions of P05362 , P19320 , P10145 , IL-1β , P08253 , P09237 , P14780 , and P24347 mRNA as well as NF-κB activation better than high dose indomethacin alone . These NSAID sparing effect of GS-HCl was further proven in collagen-induced arthritis model . Combination of GS-HCl and 2.5 mg/kg indomethacin showed significant protection from gastric damages as well as efficacious anti-arthritic effect . Taken together , P35354 N-glycosylation inhibition by GS-HCl led to indomethacin sparing effects , based on which combination of GS-HCl and reduced dose of NSAID can provide the strategy to secure stomach from NSAID-induced gastric damage as well as excellent anti-arthritic effects . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Unfavourable expression of pharmacologic markers in mucinous colorectal cancer . Patients with mucinous colorectal cancer generally have worse prognoses than those with the nonmucinous variety . The reason for this disparity is unclear , but may result from a differential response to adjuvant chemotherapy . We examined known molecular markers for response to common chemotherapy in these two histological subtypes . In all , 21 patients with mucinous and 30 with nonmucinous Dukes C colorectal cancer were reviewed for demographic data and outcome . Total RNA from the tumours and adjacent normal mucosa was isolated and reverse transcribed . Quantitative expression levels of drug pathway genes were determined using TaqMan RT-PCR ( 5-fluorouracil ( DB00544 ) : P04818 , Q12882 , P19971 ; oxaliplatin : P09211 ( glutathione S-transferase pi ) , P07992 and 2 ; irinotecan : P08183 , Q9UNQ0 , P08684 , P22309 , CES2 , P11387 ) . Mucinous tumours significantly overexpressed both P04818 and P09211 relative to nonmucinous tumours and patient-matched normal mucosa . No significant differences in expression of the remaining markers were found . Mean follow-up was 20 months ; 17 patients had recurrent disease . Among patients receiving DB00544 , those with mucinous tumours experienced shorter disease-free survival ( DFS ) than those with nonmucinous tumours ( median DFS 13.8 vs 46.5 months , P=0.053 ) . Mucinous colorectal cancer overexpresses markers of resistance to DB00544 and oxaliplatin . Likewise , DFS may be decreased in patients with mucinous tumours who receive DB00544 . The presence of mucin should be carefully evaluated in developmental trials of new agents for treating colorectal cancer . Novel combination treatments targeting chronic myeloid leukemia stem cells . Chronic myeloid leukemia ( CML ) is currently considered incurable in most patients . Stem cell transplantation , an accepted curative option for which extensive experience has been gained , is limited by high morbidity and mortality rates , particularly in older patients . Tyrosine kinase inhibitors targeting P11274 - P00519 are widely used and induce remission in a high proportion of patients , but resistance and incomplete response to these agents portends eventual relapse and disease progression . Although P11274 - P00519 inhibitors eradicate most CML cells , they are largely ineffective against the reservoir of quiescent leukemic stem cells ( LSCs ) . Thus a strong medical need exists for therapies that effectively eradicate LSCs and is currently a focus of extensive research . To date , evidence obtained from in vitro studies , animal models , and clinical CML specimens suggests that an effective approach may be to partner existing P11274 - P00519 inhibitors with compounds targeting key stem cell molecular effectors , including Wnt/β-catenin , hedgehog pathway components , histone deacetylase ( HDAC ) , transforming growth factor-β ( TGF-β ) , O60674 , promyelocytic leukemia protein , and arachidonate P09917 ( P09917 ) . Novel combinations may sensitize LSCs to P11274 - P00519 inhibitors , thereby overcoming resistance and creating the possibility of improving disease outcome beyond the current standard of care . Recombinational and physical mapping of the locus for primary open-angle glaucoma ( Q99972 ) on chromosome 1q23-q25 . Primary open-angle glaucoma ( POAG ) is a leading cause of irreversible blindness in industrialized countries . A locus for juvenile-onset POAG , Q99972 , has been mapped to 1q21-q31 in a 9-cM interval . With recombinant haplotypes , we have now reduced the Q99972 interval to a maximum of 3 cM , between the D1S452/NGA1/D1S210 and NGA5 loci . These loci are 2.8 Mb apart on a 4.7-Mb contig that we have completed between the D1S2851 and D1S218 loci and that includes 96 YAC clones and 48 STSs . The new Q99972 interval itself is now covered by 25 YACs , 30 STSs , and 16 restriction enzyme site landmarks . The lack of a NotI site suggests that the region has few CpG islands and a low gene content . This is compatible with its predominant cytogenetic location on the 1q24 G-band . Finally , we have excluded important candidate genes , including genes coding for three ATPases ( P05026 , P23634 , P50993 ) , an ion channel ( VDAC4 ) , antithrombine III ( P01008 ) , and prostaglandin synthase ( P35354 ) . Our results provide a basis to identify the Q99972 gene . DB01016 exerts an antitumor activity through reactive oxygen species-c-jun NH2-terminal kinase pathway in human gastric cancer cell line MGC-803 . DB01016 , a blocker of DB00171 -sensitive potassium ( K( DB00171 ) ) channels , can suppress progression of many cancers , but the involved mechanism is unclear . Herein we reported that MGC-803 cells expressed the K( DB00171 ) channels composed of Kir6.2 and Q09428 subunits . DB01016 induced cellular viability decline , coupled with cell apoptosis and reactive oxygen species ( ROS ) generation in MGC-803 cells . Meanwhile , glibenclamide increased NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion ( O2- ) generation , and caused mitochondrial respiration dysfunction in MGC-803 cells , suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria . DB01016 could also lead to loss of mitochondrial membrane potential , release of cytochrome c and apoptosis-inducing factor ( O95831 ) , and activation of c-jun NH2-terminal kinase ( JNK ) in MGC-803 cells . Pretreatment with antioxidant N-acetyl-l-cysteine ( Q9C000 ) prevented glibenclamide-induced JNK activation , apoptosis and cellular viability decline . Furthermore , glibenclamide greatly decreased the cellular viability , induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast ( MEF ) cells but not in P45983 -/- or P45984 -/- MEF cells . Taken together , our study reveals that glibenclamide exerts an antitumor activity in MGC-803 cells by activating ROS-dependent , JNK-driven cell apoptosis . These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer . Inhibitors of arachidonic acid metabolism reduce DNA and nuclear fragmentation induced by P01375 plus cycloheximide in U937 cells . U937 human myeloid leukemia cells are induced to apoptosis by tumour necrosis factor ( P01375 ) plus cycloheximide ( CHX ) . We have analysed the effect of various inhibitors of the arachidonic acid ( AA ) metabolism on several features of this process . The formation of high molecular weight and oligonucleosomal DNA fragments as well as nuclear fragmentation were reduced by inhibitors of P09917 ( BWA4C and BWB70C ) , P09917 activating protein ( MK-886 ) , and cytosolic P04054 ( AACOCF3 ) . None of these agents blocked the morphological changes detected by microscopy or flow cytometry , phosphatidylserine exposure on the cell surface or Caspase 3-like activation . AA also induced nuclear fragmentation at a concentration of 1-20 microM . However , the mechanisms by which these inhibitors act , remain unexplained since there was no P09917 expression in the U937 cells and no AA release followed their stimulation with P01375 plus CHX .

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Upregulation of molecules associated with T-regulatory function by thymoglobulin pretreatment of human P01730 + cells . BACKGROUND : This study evaluated the immunologic effects of thymoglobulin in modulating human P01730 + cells . METHODS : Human P01730 + cells were purified from peripheral blood mononuclear cells by negative selection method . P01730 + cells were pretreated with thymoglobulin and incubated for 72 hr . Cells and culture supernatants were collected and studied by real-time quantitative polymerase chain reaction , fluorescence activated cell scanning , multiplex cytokine assay , and mixed lymphocyte reaction ( P08235 ) . RESULTS : DB00098 pretreated P01730 + cells demonstrated up-regulation of gene transcripts for P16410 , OX40 , forkhead box P09131 ( Foxp3 ) , CD25 , P01579 , P22301 , and P60568 as determined by real-time quantitative polymerase chain reaction . Fluorescence-activated cell scanning analysis demonstrated that P01730 + cells , pretreated with thymoglobulin , up-regulated CD25 expression on their surface , and the surface expression of P16410 and OX40 and the expression of intracellular Foxp3 were observed in these P01730 +CD25+ cells . Additionally , P08235 demonstrated that thymoglobulin-pretreated cells partially inhibited proliferation of untreated autologous P01730 + cells in response to allogeneic cells . The high levels of P01579 , P22301 , P60568 , and P05112 were detected by multiplex cytokine assay in supernatants collected from cultures of thymoglobulin-pretreated P01730 + cells . The lymphocyte proliferation of allogeneic P08235 was also partially blocked in the presence of supernatants from cultures of thymoglobulin-pretreated P01730 + cells . CONCLUSIONS : This study demonstrates that the unique effects of thymoglobulin in modulating P01730 + cells may be an important mechanism for its action in inducing immunosuppression and transplant tolerance . Induction of bona fide regulatory T cells after liver transplantation - the potential influence of polyclonal antithymocyte globulin . T-cell-depleting strategies are an integral part of immunosuppressive regimens used in the hematological and solid organ transplant setting . Besides prevention of alloreactivity , treatment with rabbit antithymocyte globulin ( DB00098 ) has been related to the induction of immunoregulatory T cells ( Treg ) in vitro and in vivo . To investigate Treg induced by DB00098 , we prospectively studied the effect of DB00098 induction therapy in liver-transplanted recipients in vivo ( n = 28 ) . Treg induction was further evaluated by means of Treg-specific demethylation region ( TSDR ) analysis within the Q9BZS1 locus . Whereas no induction of P01730 (+) CD25(high) CD127(-) Treg could be observed by phenotypic analysis , we could demonstrate an induction of TSDR(+) T cells within P01730 (+) T cells exclusively for DB00098 -treated patients in the long-term ( day 540 ) compared with controls ( P = NS ) . Moreover , although in vitro experiments confirm that DB00098 primarily led to a conversion of P01730 (+) CD25(-) into P01730 (+) CD25(+) T cells displaying immunosuppressive capacities , these cells can not be classified as bona fide Treg based on their Q9BZS1 demethylation pattern . Consequently , the generation of Treg after DB00098 co-incubation in vitro does not reflect the mechanisms of Treg induction in vivo and therefore the potential clinical relevance of these cells for transplant outcome remains to be determined . Sequential phases in the development of Aire-expressing medullary thymic epithelial cells involve distinct cellular input . Intrathymic deletion of immature thymocytes that express self-reactive TCR specificities is essential in the generation of self tolerance . Medullary thymic epithelial cells ( mTEC ) expressing the transcriptional regulator Aire play a key role in this process by regulating expression of tissue-restricted antigens to ensure tolerance to peripheral tissues . Here , we have analysed the cellular and molecular requirements for the initial appearance of Aire+ mTEC in the embryonic thymus , in addition to their persistence in the adult thymus . Analysis of thymic ontogeny shows that the emergence of embryonic Aire+ mTEC occurs prior to the appearance of mature thymocytes , and depends upon lymphoid tissue inducer cells expressing retinoic acid receptor-related orphan receptor gamma . In the adult thymus , we show that Aire+ mTEC develop in the absence of thymocyte positive and negative selection and P25942 signalling , but are present at reduced frequency . Collectively these data support a model where the initial differentiation of Aire+ mTEC involves receptor activator of NF-kappaB ( Q9Y6Q6 ) - O14788 interactions with lymphoid tissue inducer cells , with subsequent mTEC turnover and/or survival involving P25942 -mediated signalling following interactions with mature P01730 + thymocytes that express P29965 . Rabbit ATG but not horse ATG promotes expansion of functional P01730 +CD25highFOXP3+ regulatory T cells in vitro . Regulatory T cells ( Treg ) play important roles in suppressing immune responses and maintaining tolerance . Rabbit antithymocyte globulin ( DB00098 ) and horse ATG ( hATG ) are widely used in the treatment of immune-mediated syndromes , but their effects on Treg are unknown . We show here that in vitro culture of normal human peripheral blood mononuclear cells ( PBMCs ) with a low-dose DB00098 resulted in marked expansion of functional Treg by converting P01730 +CD25- T cells to P01730 +CD25+ T cells . hATG did not expand but rather decreased Treg . Immuno-blot showed increased expression of Q9BZS1 and Q13469 in P01730 +CD25- and P01730 +CD25+ T cells exposed to DB00098 . PBMCs treated with DB00098 displayed increased interleukin-10 in culture supernatants than those treated with hATG . Furthermore , DB00098 and hATG showed differences in their potential to stimulate P01730 + T cells as examined using different activation markers . Microarray revealed that DB00098 induced markedly different gene-expression patterns in PBMCs , compared with hATG-treated or untreated PBMCs . Our findings indicate that DB00098 expanded Treg , probably through transcriptional regulation by enhanced Q13469 expression , in turn conferring P01730 +CD25- T cell Q9BZS1 expression and regulatory activity . The therapeutic effects of DB00098 may occur not only because of lymphocyte depletion but also enhanced Treg cell number and function . ATG-induced expression of Q9BZS1 in human P01730 (+) T cells in vitro is associated with T-cell activation and not the induction of Q9BZS1 (+) T regulatory cells . Several recent reports have suggested that in vitro exposure of P01730 (+) T cells to rabbit antithymocyte globulin ( DB00098 ) , which is commonly used to prevent and treat graft-versus-host disease and allograft rejection , is an effective method to induce P01730 (+)CD25(+) Q9BZS1 (+) T regulatory cells ( Tregs ) . We and others , however , have shown that Q9BZS1 is also expressed in activated T cells . We therefore investigated whether the induction of Q9BZS1 expression by DB00098 resulted in a stable population of suppressive Tregs . We found that exposure of peripheral blood mononuclear cells ( PBMCs ) or conventional T cells to DB00098 resulted in induction of transient rather than stable expression of CD25 and Q9BZS1 . Furthermore , DB00098 -treated T effector cells acquired neither an immunosuppressive profile of cytokine production nor suppressive capacity , even at the time of maximal Q9BZS1 expression . These findings indicate that the notion that DB00098 can be used to induce Tregs in vitro for cellular therapy in vivo should be re-evaluated . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Comparative polyclonal antithymocyte globulin and antilymphocyte/antilymphoblast globulin anti-CD antigen analysis by flow cytometry . Polyclonal antithymocyte globulin ( ATG ) /antilymphocyte and antilymphoblast globulins ( ALG ) antibodies have been used successfully in transplantation , aplastic anemia and graft-versus-host disease . Flow cytometry has been used to analyze peripheral blood lymphocyte populations in transplant patients receiving polyclonal ATG/ALG preparations for immunosuppression . Recent reports have indicated clinical dose adjustment based on levels of patient 's cells expressing various CD antigens . In vitro analysis of individual polyclonal ATG/ALG CD antigen specificity could identify appropriate antigens for clinical monitoring as well as provide useful in vitro activity data . Therefore , a flow cytometry based assay to characterize and compare activities to specific CD antigens found on the surface of peripheral blood lymphocytes has been developed . Activities found in four lots each of horse ATG ( ATGAM , Upjohn ) , rabbit and horse ATG ( thymoglobulin and lymphoglobulin , Merieux ) , horse ALG ( Minnesota ) , and DB00098 ( Fresenius ) have been compared for P06729 , CD3 , P01730 , P06127 , P09564 , CD8 , CD11a , P05107 , P10747 , P16070 , P08575 , and TCR-alpha/beta antigens . Quantitation is achieved by measuring the concentration of ATG/ALG required to give 50 % inhibition of antigen specific fluorescent-labeled monoclonal antibody relative to buffer controls . The three horse products tested have similar activity to most antigens tested . However , Fresenius DB00098 has the lowest activity for almost all antigens tested whereas the Merieux DB00098 has activities closer to the horse products . This technique allows for rapid in vitro comparison of reactivities to individual lymphocyte antigens as well as in vitro analysis of consistency . Regulatory T cells in kidney transplant recipients : the effect of induction immunosuppression therapy . BACKGROUND : Regulatory T cells have been suggested to down-regulate the alloimmune response . The aim of this prospective open study was to evaluate the effects of different inductive agents on peripheral blood regulatory T cells in kidney transplant patients and to analyse their association with short-term graft outcome . METHODS : Regulatory and effector T cell numbers in peripheral blood were determined by flow cytometry in 71 prospectively followed kidney transplant recipients at postoperative day 0 , 7 , 14 , 21 , 28 , 60 and 90 . Patients were treated with a calcineurin inhibitor-based triple immunosuppression with polyclonal rabbit anti-thymocyte globulin ( DB00098 , n = 28 ) , basiliximab , the anti-CD25 monoclonal antibody ( n = 18 ) or without induction ( controls , n = 25 ) . Flow cytometry data were correlated to rejection incidence . RESULTS : Compared to controls , P01730 (+)CD25(+)FoxP3(+) regulatory T-cell expansion among P01730 (+) T cells was noticed in the DB00098 group at all post-transplant time-points by Day 14 ( P < 0.001 ) . A significant decrease in Treg frequency ( P < 0.001 ) and concurrently a transient increase of P01730 (+)CD25(low/-)FoxP3(+) population were observed in basiliximab-treated patients 7-60 days post-transplantation . Biopsy-proven acute rejection occurred in 16.7 % of controls , 10.7 % of the DB00098 group and in 11.1 % of the basiliximab group . Higher P01730 (+)FoxP3(+)/CD8(+)CD45RA(+)CD62L(-) ratios were observed repeatedly in those patients after basiliximab induction who were rejection free ( P < 0.01 ) . CONCLUSIONS : In this study , the DB00098 induction therapy was associated with an expansion of regulatory cells . Sustained high P01730 (+)FoxP3(+)/Teff ratios were associated with the absence of rejection after basiliximab induction . Significance of immune cell function monitoring in renal transplantation after DB00098 induction therapy . Monitoring of immune status in transplant recipients is essential for predicting the risk of rejection or infection . In this study , we assessed the significance of immune cell function in 76 renal allograft recipients after DB00098 induction and initiation of maintenance immunosuppression . Using the Immuknow ( Cylex Inc ) assay , the amount of adenosine triphosphate ( DB00171 ) produced by P01730 + cells in response to phytohemagglutinin ( PHA ) was measured in patients whole blood . In parallel , the frequency and phenotype of P01730 + T cells were determined by flow cytometry . The Immuknow assay yielded paradoxically high DB00171 values during the first 3 months post-transplantation , despite very low P01730 + T cell counts . High DB00171 values were caused by peripheral blood myeloid cells , did not predict rejection , and occurred primarily in transplant recipients who received darbepoietin ( p = 0.017 ) . P01730 + T cells displayed predominantly an activated/memory phenotype and comprised a subpopulation of CD25+ Q9BZS1 + cells . Over the first 5 months post-transplantation , mean DB00171 activity gradually decreased , whereas P01730 + T cell counts slowly increased . Low DB00171 values were predictive of infection ( p = 0.002 ) . Thus Immuknow results need to be interpreted with caution in patients receiving DB00098 induction therapy . Although low DB00171 levels identify patients at increased risk for infection , high DB00171 values fail to correlate with rejection and do not justify increased immunosuppression . Antithymocyte globulins suppress dendritic cell function by multiple mechanisms . BACKGROUND : The polyclonal rabbit antithymocyte and anti-T-cell immunoglobulins ( ATGs ) DB00098 ( TG ) and DB05320 ( ATG-F ) have been widely used for the prevention and therapy of allograft rejection and graft versus host disease in transplantation . Although immunosuppressive mechanisms of ATGs on T cells are well studied , less is known about their impact on dendritic cells ( DCs ) . METHODS : Effects of TG and ATG-F on immune functions and signaling pathways of human monocyte-derived DCs were determined by flow cytometry , enzyme-linked immunosorbent assay , Western blot , apoptosis assays , endocytosis assays , and T cell stimulation assays . RESULTS : TG and ATG-F bind rapidly and with high affinity to CD11c , P33681 , P42081 , P25942 , P16671 , P28907 , CD206 , and human leukocyte antigen-DR on DCs . TG and , to a lesser extent , ATG-F induce apoptosis in immature and mature DCs . Macropinocytotic and receptor-mediated endocytotic antigen uptake in immature DCs is inhibited by TG and ATG-F due to their binding of the C-type lectins CD206 and Q9NNX6 . TG and ATG-F induce activation of the mitogen-activated protein kinases P27361 /2 and p38 that contributes to the induction of apoptosis . TG and ATG-F also induce cytoplasmic-nuclear translocation of the NF-kappaB/Rel transcription factors RelB , RelA , p50 , and p52 . Production of interleukin-12p70 in mature DCs is suppressed by TG and ATG-F . TG and ATG-F reduce the capacity of mature DCs to stimulate allogeneic and autologous T cells . CONCLUSIONS : ATGs interfere with basic DC functions , suggesting that DCs are relevant targets for the immunosuppressive action of ATGs in transplantation . Ex vivo expansion of human Tregs by DB00098 is dependent on intact P40763 -signaling in CD4⁺ T cells and requires the presence of monocytes . The addition of low , nondepleting doses of rabbit antithymocyte globulin ( ATG ) to human peripheral blood mononuclear cells has been shown to expand functional P01730 (+) CD25(+) FoxP3(+) regulatory T cells ( Tregs ) in vitro . This report is the first to elucidate the exact cellular mechanisms of ATG-mediated Treg expansion . P01730 (+) T cells require monocytes , but not other antigen presenting cell subsets , to be present in coculture to expand Tregs . However , T cells do not require direct cell-cell contact with monocytes , suggesting the importance of soluble factors . Moreover , ATG initially " reprograms " P01730 (+) T cells , but not monocytes , and induces P40763 and P42229 signaling in P01730 (+) cells . These reprogrammed P01730 (+) T cells subsequently secrete GM- P04141 and P22301 only in case of intact P40763 signaling , which in turn promote the generation of tolerogenic P08571 (+) CD11c(+) dendritic cells characterized by enhanced P22301 and decreased IL-12 production . Treg expansion following ATG treatment is accompanied by enhanced gene expression of both GM- P04141 and Bcl-2 , but not TGF-β , in peripheral blood mononuclear cells . These results demonstrate that ex vivo expansion of human Tregs by ATG is due to its ability to reprogram P01730 (+) T cells in a P40763 -dependent but TGF-β-independent manner , leading to the generation of monocyte-derived dendritic cells with a tolerogenic cytokine profile . Stimulation of host bone marrow stromal cells by sympathetic nerves promotes breast cancer bone metastasis in mice . Bone and lung metastases are responsible for the majority of deaths in patients with breast cancer . Following treatment of the primary cancer , emotional and psychosocial factors within this population precipitate time to recurrence and death , however the underlying mechanism(s) remain unclear . Using a mouse model of bone metastasis , we provide experimental evidence that activation of the sympathetic nervous system , which is one of many pathophysiological consequences of severe stress and depression , promotes MDA-231 breast cancer cell colonization of bone via a neurohormonal effect on the host bone marrow stroma . We demonstrate that induction of O14788 expression in bone marrow osteoblasts , following β2AR stimulation , increases the migration of metastatic MDA-231 cells in vitro , independently of P48061 - P61073 signaling . We also show that the stimulatory effect of endogenous ( chronic stress ) or pharmacologic sympathetic activation on breast cancer bone metastasis in vivo can be blocked with the β-blocker propranolol , and by knockdown of Q9Y6Q6 expression in MDA-231 cells . These findings indicate that O14788 promotes breast cancer cell metastasis to bone via its pro-migratory effect on breast cancer cells , independently of its effect on bone turnover . The emerging clinical implication , supported by recent epidemiological studies , is that βAR-blockers and drugs interfering with O14788 signaling , such as DB06643 , could increase patient survival if used as adjuvant therapy to inhibit both the early colonization of bone by metastatic breast cancer cells and the initiation of the " vicious cycle " of bone destruction induced by these cells . Induction of suppressive allogeneic regulatory T cells via rabbit antithymocyte polyclonal globulin during homeostatic proliferation in rat kidney transplantation . Experimental studies have shown that rabbit antithymocyte polyclonal globulin ( ATG ) can expand human P01730 +CD25++Foxp3+ cells ( Tregs ) . We investigated the major biological effects of a self-manufactured rabbit polyclonal anti-rat thymoglobulin ( DB00098 ) in vitro , as well as its effects on different peripheral T-cell subsets . Moreover , we evaluated the allogeneic suppressive capacity of DB00098 -induced Tregs in an experimental rat renal transplant model . Our results show that DB00098 has the capacity to induce apoptosis in T lymphocyte lymphocytes as a primary mechanism of T-cell depletion . Our in vivo studies demonstrated a rapid but transient cellular depletion of the main T cell subsets , directly proportional to the DB00098 dose used , but not of the effector memory T cells , which required significantly higher DB00098 doses . After DB00098 administration , we observed a significant proliferation of Tregs in the peripheral blood of transplanted rats , leading to an increase in the Treg/T effector ratio . Importantly , DB00098 -induced Tregs displayed a strong donor-specific suppressive capacity when assessed in an antigen-specific allogeneic co-culture . All of these results were associated with better renal graft function in rats that received DB00098 . Our study shows that DB00098 has the biological capacity immunomodulatory to promote a regulatory alloimmune milieu during post-transplant homeostatic proliferation . Regulatory T cells increase in breast cancer and in stage IV breast cancer . Expression levels of P15692 and Her-2 , levels of T-regulatory ( Treg ) cells , levels of CD3+ cells , and ratios of Th ( P01730 + T cells ) /Tr ( Treg ) cells were compared between stage I , II , III , and IV breast cancer patients ( n = 120 ) prior to chemotherapy and healthy women ( n = 30 ) . Cells from peripheral blood were counted by flow cytometry , Her-2 and P15692 expression was detected by pathological examination , and Her-2 was detected by Q5TCZ1 . Breast cancer patients had more Treg cells and a lower ratio of Th/Tr cells than the healthy women . Stage IV breast cancer patients had more Treg cells and a lower ratio of Th/Tr cells than stage I , II , or III breast cancer patients . Patients positive for P15692 had a lower ratio of Th/Tr cells compared with patients negative for P15692 , and those positive for both P15692 and Her-2 also had a lower ratio of Th/Tr cells compared with patients not positive for both P15692 and Her-2 . The decreased Th/Tr cells ratio indicates impaired immune function , suggesting that the stage IV breast cancer and the Her-2/ P15692 -positive breast cancer patients have lower immune function . In vivo characterization of rabbit anti-mouse thymocyte globulin : a surrogate for rabbit anti-human thymocyte globulin . BACKGROUND : Polyclonal rabbit anti-human thymocyte globulin ( DB00098 ) is used clinically for immunosuppression in solid organ transplantation ; however , it is difficult to fully characterize the effects of this agent in humans . METHODS : A surrogate rabbit anti-murine thymocyte globulin ( mATG ) was generated analogously to the commercial product DB00098 and in vivo activities were evaluated , including pharmacokinetics , T-cell depletion , dose response and kinetics , depletion/sparing of T-cell subsets or other leukocyte populations , and depletion in different lymphoid organs . RESULTS : Within 1 day , T cells are depleted by mATG in the blood , spleen , lymph node , and bone marrow down to doses of 1 mg/kg . Although mATG binds and depletes thymocytes in vitro , there is no thymocyte depletion in vivo at any dose level , suggesting decreased antibody accessibility to the thymus . After two doses of mATG given 3 days apart , T-cell reconstitution begins as early as day 9 and returns to basal levels by day 21 and 29 for P01730 and CD8 T cells , respectively . There is also preferential depletion of naïve T cells that results in increased ratios of regulatory and memory T cells within 1 day after mATG administration . Depletion of natural killer-T cells , natural killer cells , plasma cells , and plasmablasts occurs , but is modest and more transient compared with T cells . B cells , macrophages , dendritic cells , hematopoetic stem cells , and bone marrow stromal cells seem resistant to mATG depletion . CONCLUSIONS : These studies characterize the depletive effects of mATG in normal mice and provide insight into mechanisms of action of DB00098 . FoxP3+ , and not CD25+ , T cells increase post-transplant in islet allotransplant recipients following anti-CD25+ DB00098 immunotherapy . Anti-CD25 antibodies are used as an induction therapy in islet allotransplantation for type 1 diabetes . Although previous reports suggested that anti-CD25 treatment may lead to depletion of P01730 +CD25+ regulatory T cells ( Tregs ) and questioned its use in tolerance-promoting protocols for transplantation , the effect of anti-CD25 antibodies on the frequency and function of Tregs remains unclear . We examined the effect of anti-CD25 antibody , daclizumab , in vivo on Tregs in islet allograft recipients enrolled in a single-center study and monitored post-transplant . Our data shows that the reduction in CD25+ Treg cells observed post-transplant is due to masking of CD25 receptor by daclizumab and not due to depletion . In addition , using Treg marker , FoxP3 , we show that anti-CD25+ ATG treatment leads to an increase in FoxP3+ Tregs post-transplant . These data suggest that anti-CD25-based therapy has beneficial effects on Tregs and combined with ATG may be a promising therapy for autoimmunity and transplantation . Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Immune reconstitution following rabbit antithymocyte globulin . Depletional induction therapies are routinely used to prevent acute rejection and improve transplant outcome . The effects of depleting agents on T-cell subsets and subsequent T-cell reconstitution are incompletely defined . We used flow cytometry to examine the effects of rabbit antithymocyte globulin ( DB00098 ) on the peripheral T-cell repertoire of pediatric and adult renal transplant recipients . We found that while DB00098 effectively depleted CD45RA+ P26842 + naïve and CD45RO+ P26842 + central memory P01730 + T cells , it had little effect on CD45RO+ P26842 - P01730 + effector memory or CD45RA+CD31- , CD45RO+ P26842 + and CD45RO+ P26842 - CD8+ T cell subsets . When we performed a kinetic analysis of CD31+ recent thymic emigrants and CD45RA+/RO+ T cells , we found evidence for both thymopoiesis and homeostatic proliferation contributing to immune reconstitution . We additionally examined the impact of DB00098 on peripheral P01730 +Foxp3+ T cells . We found that in adults , administration of DB00098 -induced peripheral expansion and new thymic emigration of T cells with a Treg phenotype , while P01730 +Foxp3+ T cells of thymic origin predominated in children , providing the first evidence that DB00098 induces Treg in vivo . Collectively our data indicate that DB00098 alters the balance of regulatory to memory effector T cells posttransplant , providing an explanation for how it positively impacts transplant outcome . Risk factors for impaired P01730 + T-cell reconstitution following rabbit antithymocyte globulin treatment in kidney transplantation . To describe long-term P01730 + T-cell reconstitution after rabbit antithymocyte globulin ( DB00098 ) treatment and identify predictive factors following kidney transplantation . A single-center retrospective study analyzed lymphocyte subsets in DB00098 -treated kidney transplant recipients ( 1986-2009 ) . 589 patients were analyzed ( maximum follow-up 21 years ) . A comparator group ( n=298 ) received an anti- P60568 receptor monoclonal antibody . P01730 + T-cell lymphopenia ( < 200/mm3 ) was present in 48.5 % , 9.2 % , 6.7 % ,2.0 % , and 0 % of patients at one , three , five , 10 , and 20 years post-transplant , respectively . P01730 + T-cell count increased during the first 10 years but remained below the pretransplant count even after 20 years . At 1 , 3 , and 6 months post-transplant , mean P01730 + T-cell count was significantly lower in patients with P01730 + T-cell lymphopenia at 12 months versus patients without lymphopenia . On multivariate analyses , significant independent predictors for long-term impaired P01730 T-cell reconstitution were recipient age , pretransplant P01730 + T-cell count , 12-month P01730 + T-cell count , and tacrolimus or DB00688 therapy . Recipient age > 40 years was identified as a cutoff point . P01730 + T-cell reconstitution following DB00098 treatment remains impaired even after 21 years . Most risk factors for long-term impaired P01730 + T-cell reconstitution may be evaluated pretransplant or are modifiable post-transplant . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Kinetics of homeostatic proliferation and thymopoiesis after DB00098 induction therapy in kidney transplant patients . BACKGROUND : Lymphocyte-depleting therapy is associated with long-lasting effects on repopulated T cells and subsequent increased rates of infections and malignancies . The mechanisms of T-cell repopulation and their posttransplantation kinetics are not fully understood . METHODS : We studied thymopoiesis by CD31(+) naïve T cells ( recent thymic emigrants ) and homeostatic proliferation by Ki-67(+) T cells in rabbit antithymocyte globulin ( DB00098 ) -treated patients the first 6 months after transplantation . Patients receiving basiliximab or no induction therapy served as controls . RESULTS : At 6 months after transplantation , T-cell numbers were lower than before transplantation in DB00098 -treated patients , whereas T-cell numbers remained stable in both control groups . In this time period , thymopoiesis was similar between the three treatment groups ; CD8(+) T cells showed the highest percentage of recent thymic emigrants . At month 1 , percentages of Ki-67(+) naïve and memory P01730 (+) and CD8(+) T cells were the highest in DB00098 -treated patients , but these percentages declined in the months thereafter . When CD31 was used to distinguish between cytokine- and antigen-driven proliferation in naïve T cells , we found evidence for cytokine-dependent proliferation . Cytokine-dependent proliferation was also shown by in vivo increased percentages of phosphorylated P42229 and high expression levels of the interleukin-7 receptor-α and interleukin-15 receptor-α by T cells . CONCLUSION : These findings demonstrate that , in the first month after DB00098 therapy , cytokine-induced homeostatic proliferation is involved in T-cell repopulation of both naïve and memory T cells . At later time points , the contribution of homeostatic proliferation diminished , which explains the observed incomplete T-cell recovery . Polyclonal rabbit antithymocyte globulin exhibits consistent immunosuppressive capabilities beyond cell depletion . BACKGROUND : Polyclonal antithymocyte globulins ( ATGs ) are used clinically to prevent and treat acute allograft rejection and are believed to modulate the immune response primarily by depleting T cells . However , nondepleting mechanisms may also be important mediators of graft survival . In the present study , 14 lots of thymoglobulin ( DB00098 ) were analyzed and compared for nondepletive immunomodulatory activities in vitro . METHODS : Coincubation of human peripheral blood mononuclear cells with thymoglobulin induces P01730 +CD25(high)Foxp3+ regulatory T cells , which were evaluated for consistent ability to suppress T-cell activation in mixed lymphocyte reactions . The consistency of P06729 , CD3 , CD11a , and P08575 antigen specificities in thymoglobulin was determined using flow cytometry to measure inhibition of fluorescent monoclonal antibody binding to Jurkat T cells . A transwell chemotaxis assay was established and used to evaluate ATG-mediated inhibition of stromal cell-derived factor ( SDF ) -1alpha-driven Jurkat T-cell migration . RESULTS : Physiologic levels of thymoglobulin produced nondepletive immunomodulatory activities , which were consistent from batch to batch . All lots of thymoglobulin induced functionally immunosuppressive regulatory T cells and inhibited monoclonal antibody binding to key T-cell surface antigens . In addition , these studies provide the first demonstration that thymoglobulin effectively inhibits P61073 /SDF-1alpha-driven T-cell chemotaxis . CONCLUSIONS : This novel , systematic in vitro analysis of 14 different manufactured lots of thymoglobulin demonstrates the overall consistency of this product and provides further insights into nondepletive mechanisms by which thymoglobulin may generate durable immunoregulation and allograft survival . A double recombinant adenovirus expressing the costimulatory molecule P33681 -1 ( murine ) and human P60568 induces complete tumor regression in a murine breast adenocarcinoma model . Tumors that express tumor-specific antigens can maintain growth in an immunocompetent organism . Current hypotheses tend toward T cell anergy as a key component for the inhibition of immunoreactivity against such tumors . Anergy is thought to occur from hyperactive stimulation of the TCR in the absence of costimulation ( costimulation leads to proliferation via P60568 production ) . Subcutaneous injection of transgenic polyoma middle T transformed breast adenocarcinoma tumor cells ( PyMT ) in the hind flank of FVB/n mice results in the formation of tumor nodules at this site . We determined the MHC class I and class II , P33681 -1 , and P33681 -2 expression in the tumor cells by flow cytometry and showed positive staining for only MHC class I . We show that a single E1-deleted adenovirus constructed to express both the costimulatory molecule P33681 -1 ( murine ) and human P60568 genes ( Ad5E1 mB7-1/human P60568 ) elicits a very potent antitumor response when administered intratumorally . Ad5E1 mB7-1/human P60568 induced rapid and complete regression ( 100 % ) of all tumors compared with Ad5 E1 mB7-1 ( 38 % ) , Ad CAIL-2 ( 42 % ) , and Ad5E1 dl70-3 ( control vector ) ( 0 % ) . All mice that exhibited complete tumor regression were fully protected in tumor cell challenge experiments . The systemic immunity generated by intratumoral administration of the Ad vectors was associated with a strong anti-PyMT CTL response . These observations indicate that augmenting the immunogenicity of the tumor with coincident expression of P33681 -1 in combination with P60568 may prove beneficial in direct tumor immunotherapy . DB06779 , a low-molecular-weight heparin , promotes angiogenesis mediated by heparin-binding P15692 in vivo . Tumors are angiogenesis dependent and vascular endothelial growth factor-A ( P15692 ) , a heparin-binding protein , is a key angiogenic factor . As chemotherapy and co-treatment with anticoagulant low-molecular-weight heparin ( LMWH ) are common in cancer patients , we investigated whether angiogenesis in vivo mediated by P15692 is modulated by metronomic-type treatment with : ( i ) the LMWH dalteparin ; ( ii ) low-dosage cytostatic epirubicin ; or ( iii ) a combination of these two drugs . Using the quantitative rat mesentery angiogenesis assay , in which angiogenesis was induced by intraperitoneal injection of very low doses of P15692 , dalteparin sodium ( Fragmin(®) ) and epirubicin ( Farmorubicin(®) ) were administered separately or in combination by continuous subcutaneous infusion at a constant rate for 14 consecutive days . DB06779 was administered at 27 , 80 , or 240 IU/kg/day , i.e. , doses that reflect the clinical usage of this drug , while epirubicin was given at the well-tolerated dosage of 0.4 mg/kg/day . While dalteparin significantly stimulated angiogenesis in an inversely dose-dependent manner , epirubicin did not significantly affect angiogenesis . However , concurrent treatment with dalteparin and epirubicin significantly inhibited angiogenesis . The effect of dalteparin is the first demonstration of a proangiogenic effect of any LMWH in vivo . The fact that co-treatment with dalteparin and epirubicin significantly inhibited angiogenesis suggests a complex drug effect . Prolonged lymphocyte depletion by single-dose rabbit anti-thymocyte globulin and alemtuzumab in kidney transplantation . Although antibody induction has gained in popularity , two agents are rarely combined . We retrospectively analyzed peripheral lymphocyte phenotypes of renal transplant recipients who received induction therapy with a different antibody/combination : alemtuzumab(C1H) , DB00098 ( DB00098 ) , daclizumab(Dac) , DB00098 +C1H , and DB00098 +Dac . P01730 + T-cells were suppressed by C1H and DB00098 +C1H , as well as by DB00098 and DB00098 +Dac but to a lesser extent . The effect lasted for 3 years at around 40 % of baseline values . CD8+ T-cells showed a similar trend but had a more rapid recovery to baseline . P15391 + B-cells were effectively suppressed for 2 months by C1H and DB00098 +C1H , and abruptly returned to baseline afterwards ; suppression by DB00098 ( 7 doses ) was modest but lasted longer . A higher proportion of CD56+CD16+ Natural Killer cells in C1H treated patients suggested a relatively spared effect of C1H on this cell type . Low CD25+ T-cells by 5-dose Dac returned to baseline around 6 months , whereas DB00098 +C1H and DB00098 +Dac showed persistent effect . P01730 +CD25hi T-cells were suppressed by both DB00098 +C1H and DB00098 +Dac , but the initial proportion of P01730 +CD25hi T-cells among P01730 + T-cells and P01730 +CD25hi/ P01730 +CD25lo ratio were significantly higher in DB00098 +C1H . Overall , with extensive and persistent lymphocyte suppression by a simple administration of agents , single-dose DB00098 +C1H induction can be an alternative in renal transplantation . Rabbit anti T-lymphocyte globulin induces apoptosis in peripheral blood mononuclear cell compartments and leukemia cells , while hematopoetic stem cells are apoptosis resistant . Polyclonal anti-T-lymphocyte globulins ( ATG ) are used in allogeneic stem cell transplantation ( P09683 ) for the prophylaxis of graft versus host disease ( GVHD ) by in vivo T cell depletion . In this study we investigated the complement independent induction of apoptosis by DB00098 in peripheral blood mononuclear cell ( PBMNC ) compartments and hematopoetic stem cells ( P19526 ) . We also detected antileukemic activity of ATG by measuring apoptosis in myeloid and lymphatic leukemia cell lines and primary leukemia cells . We found ATG to induce apoptosis in T-lymphocytes ( P01730 (+) , CD8+ ) , B-lymphocytes ( P11836 + ) , natural killer ( NK ) -cells ( CD56(+) ) , and monocytes ( P08571 (+) ) . P19526 , in contrast , were apoptosis resistant and could be growth stimulated by low-dose ATG in the presence of bystander cells . The human leukemia cell lines Jurkat , Daudi , DG-75 ( lymphoblastic ) , and K562 , HL-60 , KG1 , and U937 ( myeloblastic ) underwent ATG-induced apoptosis , whereas the NK-cell line YT was resistant . Primary leukemia cells from 6 investigated patients with acute lymphoblastic leukemia , 9 of 10 patients with chronic lymphocytic leukemia , and 4 of 8 patients with acute myeloblastic leukemia underwent ATG-induced apoptosis . We conclude apoptosis induction in all PBMNC compartments contributes to GVHD prophylaxis . ATG might support engraftment . Finally , antileukemic activity of ATG could positively influence the transplantation outcome . Expression of cyclooxygenase-2 ( P35354 ) in tumour and stroma compartments in cervical cancer : clinical implications . This study aims at investigating the relationship between cyclooxygenase-2 expression in tumour vs stroma inflammatory compartment and its possible clinical role . The study included 99 stage IB-IV cervical cancer patients : immunostaining of tumour tissue sections was performed with rabbit antiserum against cyclooxygenase-2 . CD3 , P01730 , CD8 , CD25 , Mast Cell Tryptase monoclonal antibodies were used to characterise stroma inflammatory cells in nine cervical tumours . An inverse relation was found between cyclooxygenase-2 levels ( cyclooxygenase-2 IDV ) of tumour vs stroma compartment ( r=-0.44 , P < 0.0001 ) . The percentage of cases showing high tumour/stromal cyclooxygenase-2 IDV ratio was significantly higher in patients who did not respond to treatment ( 93.3 % ) with respect to patients with partial ( 60.5 % ) , and complete ( 43.7 % ) response ( P= 0.009 ) . Cases with a high tumour/stroma cyclooxygenase-2 IDV ratio had a shorter overall survival rate than cases with a low tumour/stroma cyclooxygenase-2 IDV ( P < 0.0001 ) . In the multivariate analysis advanced stage and the status of tumour/stroma cyclooxygenase-2 IDV ratio retained an independent negative prognostic role . The proportion of CD3(+) , P01730 (+) , and CD25(+) cells was significantly lower in tumours with high tumour/stroma cyclooxygenase-2 IDV ratio , while a higher percentage of mast cells was detected in tumours showing high tumour/stroma cyclooxygenase-2 IDV ratio . Our study showed the usefulness of assessing cyclooxygenase-2 status both in tumour and stroma compartment in order to identify cervical cancer patients endowed with a very poor chance of response to neoadjuvant therapy and unfavourable prognosis . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . DB06287 induces surfactant lipid accumulation and lung inflammation in mice . Interstitial lung disease ( ILD ) is a well-known adverse effect of mammalian target of rapamycin ( P42345 ) inhibitors . However , it remains unknown how lung toxicities are induced by P42345 inhibitors . Here , we constructed a mouse model of P42345 inhibitor-induced ILD using temsirolimus and examined the pathogenesis of the disease . Male ICR mice were treated with an intraperitoneal injection of different doses of temsirolimus ( 3 or 30 mg·kg(-1)·wk(-1) ) or vehicle . DB06287 treatment increased capillary-alveolar permeability and induced neutrophil infiltration and fibrinous exudate into the alveolar space , indicating alveolar epithelial and/or endothelial injury . It also induced macrophage depletion and the accumulation of excessive surfactant phospholipids and cholesterols . Alveolar macrophage depletion is thought to cause surfactant lipid accumulation . To further examine whether temsirolimus has cytotoxic and/or cytostatic effects on alveolar macrophages and alveolar epithelial cells , we performed in vitro experiments . DB06287 inhibited cell proliferation and viability in both alveolar macrophage and alveolar epithelial cells . DB06287 treatment caused some signs of pulmonary inflammation , including upregulated expression of several proinflammatory cytokines in both bronchoalveolar lavage cells and lung homogenates , and an increase in lymphocytes in the bronchoalveolar lavage fluid . These findings indicate that temsirolimus has the potential to induce alveolar epithelial injury and to deplete alveolar macrophages followed by surfactant lipid accumulation , resulting in pulmonary inflammation . This is the first study to focus on the pathogenesis of P42345 inhibitor-induced ILD using an animal model . Subcutaneous alemtuzumab vs ATG in adjusted conditioning for allogeneic transplantation : influence of Campath dose on lymphoid recovery , mixed chimerism and survival . Sixty-nine consecutive patients ( median age 54 years ) were prospectively enrolled in a single-institution protocol for allogeneic transplantation with adjusted non-myeloablative fludarabine-melfalan-based conditioning including cyclosporin A and DB00688 , and one of three modes of serotherapy . Thirty-one donors ( 45 % ) were unrelated . The first cohort of 29 had ATG ( DB00098 2 mg/kg x 3 days ) , the subsequent 26 had Campath 30 mg x 3 days subcutaneously , and the final cohort of 14 had 30 mg Campath once . The groups were similar as regards age , diagnosis and risk factors . Campath-patients had no acute toxicity , fewer days with fever and antibiotics , and required fewer transfusions than ATG-treated patients . 3-d-Campath patients showed lower lymphocyte counts from day +4 , and P01730 + , CD8+ , P15391 + and NK cells recovered slower than in ATG-treated patients . More Campath patients developed mixed chimerism that required DLI . 3-d-Campath induced more serious and opportunistic infections than ATG , which resulted in a greater non-relapse mortality and an impaired overall survival despite a low tumor-related mortality . The change of the Campath dosing schedule to one dose abrogated the deleterious effect of 3-d-Campath on immune recovery , severe infections and survival . Subcutaneous Campath is simple and provides strong immune suppression with no early toxicity , but dose limitation to 30 mg once is recommended . Low-dose rabbit antithymocyte globulin induction therapy results in prolonged selective lymphocyte depletion irrespective of maintenance immunosuppression . Rabbit antithymocyte globulin therapy ( DB00098 ) is a potent lymphocyte-depleting agent commonly used following renal transplantation to reduce the risk of acute rejection . Standard doses ( 7-10 mg/kg ) of DB00098 result in profound lymphopenia and predispose patients to infection and malignancy . The effects of lower doses of DB00098 ( LoD- DB00098 , 3-5 mg/kg ) on peripheral blood lymphocytes ( PBL ) are as yet unknown . In this prospective clinical trial , PBL subsets were characterized by flow cytometry over 12 months following LoD- DB00098 therapy . All patients were initially treated with standard doses of tacrolimus , mycophenolic acid , and prednisone . At 3 months , patients were randomized to either lower doses of tacrolimus or sirolimus to examine the effects of maintenance immunosuppression on PBL reemergence . LoD- DB00098 therapy resulted in prolonged suppression of P15391 + B cells , total CD3+ T cells , as well as naïve and memory P01730 + T cell and P01730 /CD25/Foxp3+ T-regulatory subsets irrespective of chronic immunosuppressive therapy . Selective depletion was only noted in the CD4CD45RA+ naïve T-cell subset resulting in an altered memory/naïve P01730 + ratio . LoD- DB00098 failed to deplete CD8+ T cells , which increased their relative contribution to the total CD3+ pool . All other lymphocyte subsets maintained near normal proportions . Thus , LoD- DB00098 therapy may lessen the adverse effects of full dose DB00098 while maintaining overall efficacy . DB00098 -associated Cd4+ T-cell depletion and infection risk in HIV-infected renal transplant recipients . HIV-infected patients are increasingly referred for kidney transplantation , and may be at an increased risk for rejection . Treatment for rejection frequently includes thymoglobulin . We studied thymoglobulin 's effect on P01730 + T-cell count , risk of infection and rejection reversal in 20 consecutive HIV-infected kidney recipients . All patients used antiretroviral therapy and opportunistic infection prophylaxis . Maintenance immunosuppression consisted of prednisone , mycophenolate mofetil and cyclosporine . Eleven patients received thymoglobulin ( 7 for rejection and 4 for delayed/slow graft function ) while 9 did not . These two groups were similar in age , gender , race , donor characteristics and immunosuppression . Mean P01730 + T-cell counts remained stable in patients who did not receive thymoglobulin , but became profoundly suppressed in those who did , decreasing from 475 +/- 192 to 9 +/- 10 cells/microL ( p < 0.001 ) . Recovery time ranged from 3 weeks to 2 years despite effective HIV suppression . Although opportunistic infections were successfully suppressed , low P01730 + T-cell count was associated with increased risk of serious infections requiring hospitalization . Rejection reversed in 6 of 7 patients receiving thymoglobulin . We conclude that thymoglobulin reverses acute rejection in HIV-infected kidney recipients , but produces profound and long-lasting suppression of the P01730 + T-cell count associated with increased risk of infections requiring hospitalization . Effects of cytokines on P15692 expression and secretion by human first trimester trophoblast cell line . PROBLEM : The mechanism through which vascular endothelial growth factor ( P15692 ) regulation occurs at the feto-maternal interface is poorly understood . The aim of this study was to investigate the effects of various cytokines on P15692 expression and secretion by trophoblast cells . METHOD OF STUDY : We investigated the effects of cytokines on P15692 expression in human first trimester trophoblast cell line by analyzing P15692 messenger RNA ( mRNA ) by reverse transcription-polymerase chain reaction and P15692 protein secretion by enzyme linked immunosorbent assay . RESULTS : The trophoblast cells expressed P15692 mRNA constitutively and the main subtypes were identified as VEGF121 and VEGF165 . When cultured in the presence of interferon ( IFN ) -gamma , interleukin ( IL ) - 1beta , tumor necrosis factor ( P01375 ) -alpha , P60568 , or P22301 , P15692 mRNA expression was found to be significantly increased by IL-1beta , P01579 and P01375 but to be unaffected by P60568 and P22301 . Moreover , P15692 secretion was most significantly increased by P01579 treatment . CONCLUSION : These results suggest that IL-1beta , P01579 , and P01375 may regulate the production of P15692 in early gestational trophoblasts . A pilot pharmacologic biomarker study of busulfan and fludarabine in hematopoietic cell transplant recipients . PURPOSE : Sixteen patients diagnosed with various hematologic malignancies participated in a phase II study evaluating the addition of rabbit antithymocyte globulin ( DB00098 , DB00098 (®) ) to the hematopoietic cell transplant ( HCT ) conditioning regimen of IV fludarabine monophosphate ( fludarabine ) and targeted intravenous ( IV ) busulfan ( fludarabine/(T)busulfan ) . Our goal was to evaluate pharmacologic biomarkers pertinent to both medications in these patients . METHODS : We characterized the interpatient variability of pharmacologic biomarkers relevant to busulfan , specifically busulfan concentration at steady state , and fludarabine , specifically F-ara-A area under the curve ( AUC ) and fludarabine triphosphate ( F-ara- DB00171 ) intracellular accumulation and concentration in separate P01730 (+) and CD8(+) T-lymphocyte populations . RESULTS : Acute and chronic graft versus host disease ( GvHD ) occurred in 11 patients and one patient , respectively . Four patients died before day +100 of non-relapse causes , which met the protocol stopping guidelines . The cumulative incidence of relapse was 25 % at 3 year post-HCT . Interpatient variability in the busulfan- and fludarabine-relevant pharmacologic biomarkers was 2.1- to 2.5-fold . F-ara-A AUC and accumulated F-ara- DB00171 in CD8(+) cells had the highest hazard ratio for non-relapse mortality and overall survival , respectively . However , neither achieved statistical significance . CONCLUSIONS : The low rates of GvHD , particularly in its chronic form , were encouraging , and further biomarker studies are warranted to optimize the fludarabine/(T)busulfan/ DB00098 conditioning regimen . DB00203 inhibits calcineurin/ Q13469 -mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin/NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase-5 ( O76074 ) inhibitor sildenafil affects calcineurin/NFAT-induced cell proliferation . MAIN METHODS : A [(3)H]thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 expressions were determined by Western blot . P24941 ( P24941 ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin/NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin/NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin/ Q13469 signaling pathway and resultant cyclin A expression , P24941 activation and cell proliferation , while the presence of DT-3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin/ Q13469 cascade-mediated cyclin A expression , P24941 activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension . Suppression of IL-12 production by soluble P29965 : evidence for involvement of the Q8TCB0 /42 mitogen-activated protein kinase pathway . IL-12 is a key cytokine in skewing immune responses toward Th1-like reactions . Human monocytes/macrophages produce high amounts of bioactive IL-12 when a priming signal ( P01579 or GM- P04141 ) precedes a second signal ( e.g. , LPS ) . We and others have previously shown that preincubation with LPS before this stimulation procedure can efficiently and selectively suppress the production of IL-12 by human monocytes . In this study , we show that an almost complete suppression of IL-12 production can also be observed after preincubation of monocytes with costimulatory cell surface molecules that bind to members of the TNFR superfamily ( P29965 , O14788 ( O14788 ) ) . The suppression of IL-12 was observable on the mRNA and protein levels and was not due to endogenous production of known IL-12 antagonists ( i.e. , P22301 , P05112 , and PGE(2) ) , to an increased number of cells undergoing apoptosis , nor to down-regulation of the P01579 or P25942 receptor . Cell surface expression of the costimulatory molecules P33681 and P42081 was not reduced by the preincubation procedure , and only a moderate reduction of P05231 production was observed . Several studies have identified signal transduction pathways that are activated by P25942 signaling , including activation of mitogen-activated protein kinases . The presence of the extracellular signal-related kinase-specific mitogen-activated protein kinase kinase 1/2-specific inhibitors PD98059 and U0126 abrogated suppression induced by sCD40 ligand or other second signals . This indicates that activation of extracellular signal-regulated kinase 1/2 contributes to the underlying mechanism of IL-12 suppression . This mechanism may be relevant in other inflammatory responses and may help to develop therapeutic strategies in Th1-mediated diseases . Interferon-gamma-induced dephosphorylation of P40763 and apoptosis are dependent on the P42345 pathway . Interferon-gamma ( P01579 ) exhibits diverse biological activities , including control of cell growth and tumor suppression . Here , we report that the treatment of M12 cells , a human metastatic prostate cancer cell line , with P01579 , resulted in marked inhibition of cell proliferation and induced apoptosis . These effects were not seen with either IFN-alpha or IFN-beta . M12 cells , like many other human cancer cells , contain constitutively activated signal transducer and activator of transcription 3 ( P40763 ) . The basal levels of both Akt and P27361 /2 phosphorylation are also markedly elevated in M12 cells . Strikingly , P01579 -induced apoptosis and growth inhibition of M12 cells were associated with persistent suppression of the constitutive tyrosine-phosphorylated P40763 ( pY- P40763 ) . The P01579 -induced dephosphorylation of pY- P40763 , however , was inhibited when the P42345 pathway was specifically blocked by rapamycin . Inhibition of PI-3K with low-dose LY294002 , or MAPK with PD98059 also suppressed the P42345 / P08133 S6k pathway , and correlated with the blockage of P01579 -induced dephosphorylation of pY- P40763 . Simultaneously , treatment with LY294002 , PD98059 , or rapamycin abolished P01579 -induced apoptosis in M12 cells . The inhibition of the P42345 pathway , however , did not affect P01579 -induced activation of P42224 pathway , and suppression of P42224 expression by siRNA had no effect on P01579 -induced dephosphorylation of pY- P40763 . Taken together , these results demonstrate that an intact P42345 pathway is critical for P01579 -induced suppression of pY- P40763 and apoptosis . Our study thus provides novel insights into the contributions of signaling pathways other than the classical JAK/ P42224 pathway in the anti-proliferative , proapoptotic actions of P01579 . Rapid T cell repopulation after rabbit anti-thymocyte globulin ( DB00098 ) treatment is driven mainly by cytomegalovirus . Rabbit anti-thymocyte globulin ( DB00098 ) induces a long-lasting lymphocytopenia . P01730 (+) T cells remain depleted for up to 2 years , whereas the CD8(+) T cell compartment is refilled rapidly by highly differentiated P26842 (-) CD45RA(+) CD57(+) effector-type cells . Because the presence of these highly differentiated CD8(+) T cells has been associated with cytomegalovirus ( CMV ) infection , we questioned to what extent restoration of CMV T cell immunity contributes to the re-emergence of T cells following DB00098 treatment . We compared T cell repopulation in six CMV-seropositive patients with CMV reactivation ( reactivating CMV(+) ) to that in three CMV(+) patients without reactivation ( non-reactivating CMV(+) ) , and to that in three CMV-seronegative recipients receiving a kidney from a CMV-seronegative donor ( CMV(-/-) ) . All patients received DB00098 because of acute allograft rejection . Total P01730 and CD8 counts , frequency and phenotype of virus-specific CD8(+) T cells were determined . In reactivating CMV(+) patients , total CD8(+) T cells reappeared rapidly , whereas in non-reactivating CMV(+) patients they lagged behind . In CMV(-/-) patients , CD8(+) T cell counts had not yet reached pretransplant levels after 2 years . CMV reactivation was indeed followed by a progressive accumulation of CMV-specific CD8(+) T cells . During lymphocytopenia following DB00098 treatment , serum interleukin ( IL ) -7 levels were elevated . Although this was most prominent in the CMV-seronegative patients , it did not result in an advantage in T cell repopulation in these patients . Repopulated CD8(+) T cells showed increased skewing in their Vβ repertoire in both CMV(-/-) and reactivating CMV-seropositive patients . We conclude that rapid T cell repopulation following DB00098 treatment is driven mainly by CMV . DB00472 induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5-HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase-2 ( P35354 ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg(-1) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 ; i.p. , 125mgkg(-1) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5-HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5-hydroxyindoleacetic acid ( 5-HIAA ) levels ( P < 0.01 ) and , P28335 receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 ( P < 0.001 ) , and P35354 expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5-HT metabolism and/or its ability to reduce colonic malignant events . [ Renal cell carcinoma management and therapies in 2010 ] . Advanced renal cell carcinoma is associated with a poor prognosis and is refractory to standard chemotherapy . Recent progress in the understanding of molecular biology and pathogenesis of renal cell cancer has been translated into the development of new therapeutic strategies . The management of metastatic RCC has been revolutionized with the development of targeted molecular therapies against P15692 -VEGFR and P42345 . Randomized phase III clinical trials demonstrated clinical benefit for patients with advanced RCC in overall survival and progression free survival . At the moment , six molecules have been approves in advanced RCC : cytokines ( P60568 and IFN ) , antiangiogenic therapies ( sunitinib , sorafenib , bevacizumab ) and P42345 inhibitors ( DB06287 , everolimus ) . Nephrectomy is an important component of the multimodality treatment of mRCC . Prospective trials will be assessed the value of nephrectomy in patients treated by antiangiogenic therapies . Large randomized trial are ongoing to evaluate these new therapies in adjuvant setting . In vitro cell death of activated lymphocytes in Omenn 's syndrome . Omenn 's syndrome ( OS ) is characterized by erythrodermia , hepatosplenomegaly , lymphadenopathy , hypereosinophilia and elevated IgE levels associated with increased susceptibility to severe infections . Peripheral blood T cells , though usually present in normal number , show an activated phenotype ( including an increased expression of CD95/Fas ) , a Th2 pattern of cytokine secretion and defective proliferative response to mitogens . In this report , we demonstrate that T cells from patients with OS undergo an excessive cell death in vitro resulting from two mechanisms . First , a substantial number of peripheral blood lymphocytes from OS patients die in unstimulated cultures ( p = 0.009 vs. healthy controls ) . This spontaneous apoptosis is associated with reduced expression of bcl-2 gene product ( p < 0.05 ) and can be prevented by addition of interleukin ( IL ) -2 ( which also prevents down-modulation of bcl-2 ) , while is independent from CD95 signaling . Second , lymphocytes from OS patients are highly susceptible to activation-induced cell death ( AICD ) induced with mitogens . This mechanism is largely independent from P60568 , while it can be significantly inhibited blocking CD95 with an IgG2b monoclonal antibody ( mAb ) . The dependence of AICD from signals transduced via CD95 was confirmed showing that cross-linking CD95 with an IgM mAb induces a higher cell death in purified P01730 + CD45R0+ cells from OS patients than in controls ( comparable for CD95 expression ) . Both mechanisms of cell death observed in this study result from lymphocyte hyperactivation occurring in vivo in these patients and may contribute to functional T cell defects of OS . Rabbit anti-rat thymocyte immunoglobulin preserves renal function during ischemia/reperfusion injury in rat kidney transplantation . Ischemia/reperfusion ( I/R ) injury is an important cause of renal graft dysfunction in humans . Increases in cold and warm ischemia times lead to a higher risk of early post-transplant complications including delayed graft function and acute rejection . Moreover , prolonged cold ischemia is a predictor of long-term kidney graft loss . The protective effect of rabbit anti-rat thymocyte immunoglobulin ( DB00098 ) was evaluated in a rat model of I/R injury following syngeneic kidney transplantation . Serum creatinine concentration was evaluated at 16 h and 24 h post-transplant . Animals were sacrificed 24 h post-transplant for evaluation of histology , infiltrating leukocytes , nitrotyrosine staining , and apoptosis . DB00098 was effective in preventing renal function impairment , tissue damage and tubular apoptosis associated with I/R only when was given 2 h before transplantation but not at the time of reperfusion . Pretransplant DB00098 treatment of recipient animals effectively reduced the amount of macrophages , P01730 (+) , CD8(+) T cells and LFA-1(+) cells infiltrating renal graft subjected to cold ischemia as well as granzyme-B expression within ischemic kidney . On the other hand , granulocyte infiltration and oxidative stress were not modified by DB00098 . If these results will be translated into the clinical setting , pretransplant administration of Thymoglobuline(®) could offer the additional advantage over peri-transplant administration of limiting I/R-mediated kidney graft damage . A cell-intrinsic inhibitor of HIV-1 reverse transcription in P01730 (+) T cells from elite controllers . HIV-1 reverse transcription represents the predominant target for pharmacological inhibition of viral replication , but cell-intrinsic mechanisms that can block HIV-1 reverse transcription in a clinically significant way are poorly defined . We find that effective HIV-1 reverse transcription depends on the phosphorylation of viral reverse transcriptase by host cyclin-dependent kinase ( CDK ) 2 at a highly conserved DB00156 residue . P24941 -dependent phosphorylation increased the efficacy and stability of viral reverse transcriptase and enhanced viral fitness . Interestingly , P38936 , a cell-intrinsic CDK inhibitor that is upregulated in P01730 (+) T cells from " elite controllers , " potently inhibited P24941 -dependent phosphorylation of HIV-1 reverse transcriptase and significantly reduced the efficacy of viral reverse transcription . These data suggest that P38936 can indirectly block HIV-1 reverse transcription by inhibiting host cofactors supporting HIV-1 replication and identify sites of viral vulnerability that are effectively targeted in persons with natural control of HIV-1 replication . The effect of rabbit antithymocyte globulin on human DB05914 . Mesenchymal stem cells ( MSCs ) possess immunomodulatory properties which are of key interest for their application in autoimmunity and transplantation . In transplantation , administration of MSCs has shown promising results in preclinical models and has recently moved to clinical trials . Therefore , it is important to study the interactions between MSCs and immunosuppressive drugs currently used in transplantation . We aimed to analyze the effect of rabbit antithymocyte globulin ( DB00098 ) MSCs . MSCs were obtained from perirenal fat of kidney donors and exposed to ranging doses of DB00098 ( DB00098 (®) , Genzyme ; 0.5-100 μg/ml ) . Binding of DB00098 , effects on viability and susceptibility to be killed by cytotoxic lymphocytes as well as effects on their immunosuppressive potential of MSCs were tested . DB00098 binds dose-dependently to MSCs . This binding was associated with slightly impaired viability after 48 and 72 h when compared with nonexposed MSCs . In contrast to nontreated MSCs , DB00098 preexposed MSCs were susceptible to be lysed by cytokine-activated CD8(+) cytotoxic cells and NKT cells . The capacity of MSCs to suppress the proliferation of anti-CD3/ P10747 activated P01730 and CD8 T cells were reduced by the presence of DB00098 in the culture . DB00098 reduces the viability and antiproliferative capacity of MSCs in a dose-dependent manner and converts them into targets for CD8 T cells and NKT cell lysis . Murine antithymocyte globulin T-cell depletion is mediated predominantly by macrophages , but the Fas/ P48023 pathway selectively targets regulatory T cells . BACKGROUND : DB00098 is a T-cell-depleting polyclonal rabbit anti-human thymocyte antibody used clinically for immunosuppression in solid organ and hematopoietic stem-cell transplantation . By using a surrogate rabbit anti-mouse thymocyte globulin ( mATG ) , we previously demonstrated that murine regulatory and memory T cells are preferentially spared from mATG depletion in vivo . The current studies were designed to determine whether different effector mechanisms are involved in differential depletion of T-cell subsets by mATG . METHODS : Complement-dependent cytotoxicity , antibody-dependent cellular cytotoxicity ( ADCC ) , and apoptotic mechanisms of depletion by mATG were evaluated in vitro and in vivo . RESULTS : In vitro , there was evidence of differential susceptibility of T-cell subsets by different effector mechanisms where naïve and P01730 effector memory T cells show reduced susceptibility to apoptosis , whereas regulatory T cells are less susceptible to mATG-mediated complement-dependent cytotoxicity and ADCC . However , mATG treatment of mice depleted of ADCC effector cell types ( neutrophils , natural killer cells , or macrophages ) or deficient in complement P01031 or Fas demonstrated that mATG depletion of all T-cell subsets is mediated primarily by macrophages and that the role of neutrophils , natural killer cells , and complement is minimal in vivo . Interestingly , the Fas/ P48023 pathway does play a role in regulatory T-cell depletion , which is likely a result of increased basal expression of Fas on these cells . CONCLUSIONS : These data suggest that macrophages deplete most T cells by mATG in mice , but regulatory T cells are also uniquely susceptible to mATG-mediated Fas-dependent depletion . Opposed effects of lithium on the MEK- P29323 pathway in neural cells : inhibition in astrocytes and stimulation in neurons by GSK3 independent mechanisms . DB01356 is widely used in the treatment of bipolar disorder , but despite its proven therapeutic efficacy , the molecular mechanisms of action are not fully understood . The present study was undertaken to explore lithium effects of the MEK/ P29323 cascade of protein kinases in astrocytes and neurons . In asynchronously proliferating rat cortical astrocytes , lithium decreased time- and dose-dependently the phosphorylation of MEK and P29323 , with 1 mM concentrations achieving 60 and 50 % inhibition of P29323 and MEK , respectively , after a 7-day exposure . DB01356 also inhibited [3H]thymidine incorporation into DNA and induced a G2/M cell cycle arrest . In serum-deprived , quiescent astrocytes , pre-exposure to lithium resulted in the inhibition of cell cycle re-entry as stimulated by the mitogen endothelin-1 : under this experimental setting , lithium did not affect the rapid , peak phosphorylation of MEK taking place after 3-5 min , but was effective in inhibiting the long-term , sustained phosphorylation of MEK . DB01356 inhibition of the astrocyte MEK/ P29323 pathway was independent of inositol depletion . Further , compound SB216763 inhibited Tau phosphorylation at Ser396 and stabilized cytosolic beta-catenin , consistent with the inhibition of glycogen synthase kinase-3 beta ( P49841 ) , but failed to reproduce lithium effects on MEK and P29323 phosphorylation and cell cycle arrest . In cerebellar granule neurons , millimolar concentrations of lithium enhanced MEK and P29323 phosphorylation in a concentration-dependent manner , again through an inositol and P49841 independent mechanism . These opposing effects in astrocytes and neurons make lithium treatment a promising strategy to favour neural repair and reduce reactive gliosis after traumatic injury . The effect of different ATG preparations on immune recovery after allogeneic hematopoietic stem cell transplantation for severe aplastic anemia . Anti-thymocyte globulin ( ATG ) is widely used in the conditioning regimen before allogeneic stem cell transplantation for aplastic anemia . However , there are several different preparations of ATG and little is known about the difference of their effects on transplantation outcome . Therefore , in this study , we retrospectively compared the effect of two different DB00098 preparations [ DB00098 ( ATG-G ) and ATG-Fresenius ( ATG-F ) ] on immune recovery and cytomegalovirus infection after transplantation . The conditioning regimen was a combination of fludarabine , cyclophosphamide , and ATG . Low dose total body irradiation was added in alternative donor transplantation . Four patients received ATG-F at 5 mg/kg/day from day -7 to day -3 , whereas ATG-G was given at 2.5 mg/kg/day from day -5 to day -2 in three patients . There was no graft rejection and no grade II-IV acute graft-versus-host disease . All three patients in the ATG-G group developed positive cytomegalovirus antigenemia including two with high-grade antigenemia , whereas two of the four patients in the ATG-F group were persistently negative . Immunological evaluation on day 60 revealed that both P01730 + and CD8+ T-cell recoveries were delayed in the ATG-G group . These findings suggested that ATG-G has a stronger immunosuppressive activity than the ATG-F with a dose ratio of 1:2.5 . Acute and chronic vascular rejection in nonhuman primate kidney transplantation . A nonhuman primate ( NHP ) study was designed to evaluate in nonlife-supporting kidney allografts the progression from acute rejection with transplant endarteritis ( TXA ) to chronic rejection ( CR ) with sclerosing vasculopathy . Group P55008 ( n = 6 ) received high cyclosporine A ( DB00091 ) immunosuppression and showed neither TXA nor CR during 90 days post-transplantation . Group G2 ( n = 6 ) received suboptimal DB00091 immunosuppression and showed severe TXA with graft loss within 46 days ( median ) . Arterial intimal changes included infiltration of macrophages and T lymphocytes ( CD3 , P01730 , CD8 ) with few myofibroblasts , abundant fibronectin/collagen IV , scant collagens I/III , high rate of cellular proliferation and no C4d accumulation along peritubular capillaries . Group P46379 ( n = 12 ) received suboptimal DB00091 and anti-rejection therapy ( DB00098 + methylprednisolone + DB00091 ) of TXA . Animals developed CR and lost grafts within 65 days ( median ) . As compared to G2 , the arterial intimal changes showed less macrophages and T lymphocytes , an increased number of myofibroblasts , abundant fibronectin/collagen IV and scar collagens I/III , C4d deposition along capillaries in 60 % of animals and transplant glomerulopathy in 80 % of animals . In conclusion , CR is an immune stimulated process initiated during TXA with the accumulation and proliferation of myofibroblasts , and progressive deposition of collagens in the intima . Our experimental design appears well suited to study events leading to CR . Atypical acute rejection after hand transplantation . Skin rejection after hand transplantation is characterized by a maculopapular erythematous rash that may be diffuse , patchy or focal , and distributed over forearms and dorsum of the hands . This ' classical ' pattern of rejection usually spares the skin of the palm and does not affect the nails . Herein , we report the experience on four cases presenting with an ' atypical ' pattern of rejection that is novel in involving the palmar skin and the nails . All patients were young and exposed to repetitive and persistent mechanical stress of the palm . Characteristic features of rejection included a desquamative rash associated with dry skin , red papules , scaling and lichenification localized to the palm . Skin lesions were associated with nail dystrophy , degeneration , deformation or loss . Histology of the skin and nail bed revealed a lymphocytic infiltrate with predominance of T cells ( CD3+ , P01730 + and CD8+ ) , with small numbers of B cells ( P11836 + and CD79a+ ) and a low number of Forkhead transcription factor 3 ( Q9BZS1 ) -positive cells in one patient . The lesions persisted over weeks to months , responded poorly to steroid treatment and were managed with antithymocyte globulin ( ATG ; DB00098 , Genzyme , Cambridge , MA ) , alemtuzumab and/or intensified maintenance immunosuppression . Selective serotonin reuptake inhibitors attenuate the antigen presentation from dendritic cells to effector T lymphocytes . DB00472 , one of the selective serotonin reuptake inhibitors ( SSRIs ) , has been found to possess immune modulation effects , in addition to its antidepressant effects . However , it remains unclear whether SSRIs can suppress the antigen-presenting function of dendritic cells ( DCs ) . Therefore , DB00472 was applied to a co-culture of Aggregatibacter actinomycetemcomitans ( Aa ) -reactive T cells ( ×Aa-T ) isolated from Aa-immunized mice and DCs . This resulted in the suppressed proliferation of ×Aa-T stimulated with Aa-antigen presentation by DCs . Specifically , DB00472 increased the extracellular 5-hydroxytryptamine ( 5-HT ) in the ×Aa-T/DC co-culture , whereas exogenously applied 5-HT promoted T-cell proliferation in the ×Aa-T/DC co-culture , indicating that DB00472 -mediated suppression of ×Aa-T/DC responses can not be attributed to extracellular 5-HT . Instead , DB00472 remarkably suppressed the expression of costimulatory molecule Q9Y6W8 -L on DCs . DB00472 also promoted a greater proportion of P42081 (Low) immature DCs than P42081 (High) mature DCs , while maintaining the expression levels of P33681 , MHC-class-II and Q9NZQ7 . These results suggested that DB00472 suppressed the ability of DCs to present bacterial antigens to T cells , and the resulting T-cell proliferation , in a P31645 /5-HT-independent manner and that diminished expression of Q9Y6W8 -L on DCs and increase of P42081 (Low) immature DCs caused by DB00472 might be partially associated with DB00472 -mediated suppression of DC/T-cell responses . Human P01730 (+)CD25(+) Cells in Combination with P28906 (+) Cells and DB00098 to Prevent Anti-hematopoietic Stem Cell T Cell Alloreactivity . Cotransplantation of human P28906 (+) hematopoietic stem cells ( P19526 ) and P01730 (+)CD25(+)FoxP3(+) regulatory T cells ( Tregs ) could prevent anti- P19526 alloreactivity and reduce the risk of rejection in HLA mismatched transplants . To pursue this hypothesis we cocultured P28906 (+) cells and P01730 (+)CD25(+) cells immunomagnetically isolated ( Milteny ) from human peripheral blood ( unmanipulated or granulocyte-colony stimulating factor [ DB00099 ] mobilized ) or cord blood . Enriched Tregs obtained from the same source ( autologous ) of P28906 (+) cells showed greater inhibitory effect on T cell alloreactivity than third-party ( allogeneic ) Tregs . The immunosuppressive activity of Tregs was maintained after stimulation with allogeneic P28906 (+) cells and Tregs did not modify the clonogenic activity of P28906 (+) cells in vitro . Cotransplantation of Tregs with P28906 (+) cells at 1:1 or 2:1 ratios in nonobese diabetic/severe combined immunodeficiency ( NOD/SCID ) mice resulted in normal hematopoietic stem cell engraftment . Incubation with physiologic doses of rabbit antithymocyte globulin ( DB00098 , thymoglobulin ) did not affect the number of Tregs in 6-day culture . Upon exposure to thymoglobulin Tregs maintained their suppressive activity , increased expression of P32248 , and released multiple cytokines , primarily interleukin (IL)10 . Our findings suggest that human autologous or allogeneic Tregs could be cotransplanted with P28906 (+) cells after preparative regimens including thymoglobulin . DB01356 inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells . BACKGROUND : Exposing eukaryotic cells to lithium ions ( Li+ ) during development has marked effects on cell fate and organization . The phenotypic consequences of Li+ treatment on Xenopus embryos and sporulating Dictyostelium are similar to the effects of inhibition or disruption , respectively , of a highly conserved protein serine/threonine kinase , glycogen synthase kinase-3 ( GSK-3 ) . In Drosophila , the GSK-3 homologue is encoded by zw3sgg , a segment-polarity gene involved in embryogenesis that acts downstream of wg . In higher eukaryotes , GSK-3 has been implicated in signal transduction pathways downstream of phosphoinositide 3-kinase and mitogen-activated protein kinases . RESULTS : We investigated the effect of Li+ on the activity of the GSK-3 family . At physiological doses , Li+ inhibits the activity of human P49841 and Drosophila Zw3Sgg , but has no effect on other protein kinases . The effect of Li+ on GSK-3 is reversible in vitro . Treatment of cells with Li+ inhibits GSK-3-dependent phosphorylation of the microtubule-associated protein Tau . Li+ treatment of Drosophila S2 cells and rat PC12 cells induces accumulation of cytoplasmic Armadillo/beta-catenin , demonstrating that Li+ can mimic Wingless signalling in intact cells , consistent with its inhibition of GSK-3 . CONCLUSIONS : Li+ acts as a specific inhibitor of the GSK-3 family of protein kinases in vitro and in intact cells , and mimics Wingless signalling . This reveals a possible molecular mechanism of Li+ action on development and differentiation . Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma . Tumor-derived exosomes contain biologically active proteins and messenger and microRNAs ( miRNAs ) . These particles serve as vehicles of intercellular communication and are emerging mediators of tumorigenesis and immune escape . Here , we isolated 30-100 nm exosomes from the serum of patients with nasopharyngeal carcinoma ( NPC ) or the supernatant of TW03 cells . Increased circulating exosome concentrations were correlated with advanced lymphoid node stage and poor prognosis in NPC patients ( P < 0.05 ) . TW03-derived exosomes impaired T-cell function by inhibiting T-cell proliferation and Th1 and Th17 differentiation and promoting Treg induction by NPC cells in vitro . These results are associated with decreases in P29323 , P42224 , and P40763 phosphorylation and increases in P42229 phosphorylation in exosome-stimulated T-cells . TW03-derived exosomes increased the proinflammatory cytokines IL-1β , P05231 , and P22301 but decreased IFNγ , P60568 , and Q16552 release from P01730 + or CD8+ T-cells . Furthermore , five commonly over-expressed miRNAs were identified in the exosomes from patient sera or NPC cells : hsa-miR-24-3p , hsa-miR-891a , hsa-miR-106a-5p , hsa-miR-20a-5p , and hsa-miR-1908 . These over-expressed miRNA clusters down-regulated the Q9P0L2 signaling pathway to alter cell proliferation and differentiation . Overall , these observations reveal the clinical relevance and prognostic value of tumor-derived exosomes and identify a unique intercellular mechanism mediated by tumor-derived exosomes to modulate T-cell function in NPC . Increased levels of Candida albicans mannan-specific T-cell-derived antigen binding molecules in patients with invasive candidiasis . In addition to cytokines , P01730 + T cells have been found to secrete soluble , T-cell-derived antigen binding molecules ( TABMs ) . These antigen-specific immunoproteins are thought to have immunoregulatory properties in the suppression of cell-mediated immunity ( CMI ) because they often associate with interleukin-10 ( P22301 ) and transforming growth factor beta . Decreased CMI causes susceptibility to infections caused by organisms which are normally nonpathogenic . In this situation , e.g. , Candida albicans saprophytism may develop into invasive candidiasis . The difficult diagnosis of invasive candidiasis is based on the findings obtained from blood cultures and with tissue biopsy specimens , with some additional diagnostic value gained by the detection of Candida albicans mannan antigenemia and antimannan antibodies . In the present study , Candida albicans mannan-specific TABM ( P62158 -TABM ) levels in the sera of patients with invasive candidiasis ( n = 11 ) , Candida colonization ( n = 11 ) and noncolonization ( n = 10 ) , recurrent vulvovaginal candidiasis ( n = 30 ) , and atopic eczema dermatitis syndrome ( n = 59 ) and healthy controls ( n = 30 ) were analyzed . For 14 participants , the effect of mannan stimulation on TABM production and gamma interferon ( P01579 ) and P05112 mRNA expression by peripheral blood lymphocytes was also studied . It was demonstrated that P62158 -TABM production was the highest in patients with invasive candidiasis and that P62158 -TABM levels could distinguish Candida-colonized patients from noncolonized patients . In addition , the P62158 -TABM level was directly related to mRNA expression for P05112 but not P01579 . These results reinforce the view that TABMs are associated with decreased CMI , immunoregulation , and the T-helper cell 2-type immune response . Naive P01730 t cell proliferation is controlled by mammalian target of rapamycin regulation of Q8TEB7 expression . In this study , we demonstrate that the E3 ubiquitin ligase gene related to anergy in lymphocytes ( Q8TEB7 ) is expressed in quiescent naive mouse and human P01730 T cells and has a functional role in inhibiting naive T cell proliferation . Following TCR engagement , P10747 costimulation results in the expression of P60568 whose signaling through its receptor activates the Akt-mammalian target of rapamycin ( P42345 ) pathway . Activation of P42345 allows selective mRNA translation , including the epistatic regulator of Q8TEB7 , Q96FW1 ( Otub1 ) , whose expression results in the degradation of Q8TEB7 and allows T cell proliferation . The activation of P42345 appears to be the critical component of IL-2R signaling regulating Q8TEB7 expression . P16410 -Ig treatment blocks P10747 costimulation and resultant P60568 expression , whereas rapamycin and anti- P60568 treatment block P42345 activation downstream of IL-2R signaling . Thus , all three of these biotherapeutics inhibit P42345 -dependent translation of mRNA transcripts , resulting in blockade of Otub1 expression , maintenance of Q8TEB7 , and inhibition of P01730 T cell proliferation . These observations provide a mechanistic pathway sequentially linking P10747 costimulation , IL-2R signaling , and P42345 activation as important requirements for naive P01730 T cell proliferation through the regulation of Otub1 and Q8TEB7 expression . Our findings also extend the role of Q8TEB7 beyond anergy induction and maintenance , suggesting that endogenous Q8TEB7 regulates general cell cycle and proliferation of primary naive P01730 T cells . SV40 infection associated with rituximab treatment after kidney transplantation in nonhuman primates . BACKGROUND : A regimen consisting of polyclonal anti-T-cell antibody , sirolimus ( Q86TD4 ) , and donor bone marrow ( DBM ) infusion induces robust transplantation tolerance to skin allografts in mice . We investigated the effect of a similar regimen in a nonhuman primate ( NHP ) model . METHODS : Cynomolgus macaques ( Macaca fascicularis ) were transplanted with mismatched kidney allografts . Recipients were treated with 7 doses of antithymocyte globulin ( DB00098 , day 1 to 9 ) , sirolimus , and DBM infusion ( day 14 ) . Anti- P11836 antibody , rituximab , was given on days 0 and 5 . RESULTS : A regimen of DB00098 , 30 days of Q86TD4 , and DBM infusion induced significantly greater prolongation of graft survival with a mean survival time of 88 days compared with the control regimen ( no DBM ) with an mean survival time of 53 days ( P=0.022 ) . Unlike the murine skin allograft model , all grafts were rejected within 111 days . A combination of DB00098 , continuous Q86TD4 , and rituximab caused graft and systemic SV40 infection and failed to achieve further extension of graft survival . C4d deposition was observed in 50 % of recipients as early as 18 days , suggesting antidonor antibody production . A transient , low-to-moderate degrees of multilineage chimerism was observed after DBM infusion . Treatment with DB00098 resulted in profound depletion of P01730 + and CD8+ T cells , whereas addition of rituximab achieved prolonged ( up to 3 months ) depletion of P11836 + B cells . CONCLUSION : The DB00098 , Q86TD4 , and DBM protocol is simple and produces long-term kidney allograft survival in NHP although additional treatment modalities may be necessary for induction of long-term tolerance . ATG induction in renal transplant recipients : Long-term hazard of severe infection is associated with long-term functional T cell impairment but not the ATG-induced P01730 cell decline . BACKGROUND AND METHODS : We showed previously that DB00098 induction induces a strong decrease of P01730 + T cells together with impaired in vitro P60568 secretion up to 1 year post-transplant . To further characterize long-term immunological effects of ATG induction 2 and 5 years post-transplant , we used sensitive intracellular cytokine analysis in the same prospective study of 84 renal transplant recipients ( ATG , n=44 ) . RESULTS : A significantly increased frequency of severe infectious disease ( HR=2.0 , p=0.027 ) as well as suppressed T cell functions were found within 2 years after ATG induction but not beyond ( logistic regression ( logreg ) : P01730 cell P22301 responses , p=0.064 ; T cell proliferation , p=0.038 ) . Impaired T cell proliferation at 2 years was associated with occurrence of severe infection ( p=0.017 ) . Importantly , a strong and persistent decrease of P01730 cell counts ( p < 0.0005 at 5 years ) was independently associated with ATG induction ( logreg p=0.002 ) but not related to functional P01730 cell impairment ( helper activity/cytokine production ) or an increased risk of infection . CONCLUSIONS : Severe infection up to 2 years after ATG induction was associated with impaired T cell proliferative capacity but not with the profound decline in P01730 cell counts that occurred after ATG induction and persisted up to 5 years . Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P. bainier 229-7 mycelia was increased in response to exposure to high external Ca(2+) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca(2+) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72-h biotransformation . The Ca(2+) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72-h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 . These results suggest that both Ca(2+) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi . Preferential increase in memory and regulatory subsets during T-lymphocyte immune reconstitution after DB00098 induction therapy with maintenance sirolimus vs cyclosporine . BACKGROUND : DB00877 maintenance therapy with DB00098 induction is a promising regimen that may preserve renal function . Data are lacking , however , about the immunologic effects of combined DB00098 -sirolimus . METHODS : In a 12-month , prospective , randomised , open-label , single-centre pilot study , de novo deceased-donor kidney transplant patients were randomised to receive cyclosporine or sirolimus , with DB00098 induction , mycophenolate mofetil and corticosteroids . Flow cytometry analysis of peripheral blood was used to evaluate immune reconstitution . RESULTS : Nineteen patients were recruited ( sirolimus 9 , cyclosporine 10 ) . Reconstitution of the P01730 (+) T-lymphocyte subset was significantly lower with sirolimus versus cyclosporine over year 1 , but CD8(+) reconstitution did not differ significantly between groups . The proportion of naïve P01730 (+) T-lymphocytes showed an initial decrease with sirolimus versus cyclosporine . Naïve CD8(+) T-lymphocytes increased versus baseline in the cyclosporine cohort at months 1 and 3 , but remained unchanged with sirolimus . Memory P01730 (+) T-lymphocytes occurred more frequently in sirolimus- versus cyclosporine-treated patients during year 1 . The proportion of memory CD8(+) T-lymphocytes decreased at months 1 and 3 compared to baseline in the DB00091 arm , but did not change in the sirolimus cohort . By month 12 , the proportion of both naïve and memory P01730 (+) and CD8(+) T-lymphocytes had become similar with sirolimus or cyclosporine . There were fewer naïve B-lymphocytes in the sirolimus cohort and more P15391 (-)IgD(+/-) P26842 (+) memory B-lymphocytes . CONCLUSIONS : In this small population , homeostatic reconstitution after DB00098 induction showed disproportionately high recovery of memory T-lymphocyte subsets during sirolimus therapy , which may explain the higher rejection rate seen with sirolimus versus cyclosporine following kidney transplantation . Effect of different induction strategies on effector , regulatory and memory lymphocyte sub-populations in clinical islet transplantation . This prospective study assessed lymphocyte subsets in the peripheral blood of 42 islet allograft recipients using flow cytometry from 2 weeks and up to 2 years post-transplantation . Subjects received daclizumab ( n = 16 ) , DB00098 ( n = 12 ) or alemtuzumab ( n = 14 ) . DB00087 was associated with an early ( within 1 month ) and transient ( up to 6 months ) increase in the frequency of CD3(+) P01730 (+) Foxp3(+) T cells , while daclizumab induced a near complete loss of these cells ( P < or= 0.001 ) . The frequency of memory P01730 (+) T cells was increased following depleting immunosuppression induction with either DB00098 or alemtuzumab ( P < or= 0.05 ) , but remained unchanged while using daclizumab . DB00087 induction resulted in a significant loss of memory B lymphocytes when compared with the other induction groups ( P < or= 0.001 ) . While the clinical significance of these findings remains to be fully determined , the observed altered balance between effector , regulatory and memory cells suggests that the immune status of patients will be affected according to the induction strategy chosen . Effect of thymoglobulin induction on HIV-infected renal transplant recipients : differences between HIV-positive and HIV-negative patients . The best immunosuppressive regimen in HIV-infected renal transplant recipients has not been established . DB00098 has been associated with an increased risk of serious bacterial infections in HIV-negative patients and , for this reason , there is some concern over its use in the HIV-infected population . We describe three consecutive HIV-infected renal transplant recipients who received thymoglobulin as induction therapy , and we compared their progress with a cohort of 23 HIV-negative recipients . Median follow-up was 24 and 11 months , respectively . Nadir lymphocytopenia was observed at 1 week in both groups , and their absolute lymphocyte count recovery was similar . An early and deep ( < 30 cells/mm(3) ) P01730 (+) T cell lymphocytopenia was seen in two of the three HIV-infected patients . No opportunistic infections were diagnosed in HIV-positive patients . One HIV-positive patient had a bacterial infection and five HIV-negative patients had one or more bacterial infections . DB00098 was safe in our three HIV-infected renal transplant recipients . Until those data are confirmed in larger studies , close monitoring is recommended during the thymoglobulin-induced P01730 (+) T cell lymphocytopenia period . Acute cellular rejection . The incidence of acute rejection of the kidney allograft in the world has been around 15 % during the period between 2001 and 2003 . It is clinically defined as an elevation in the level of serum creatinine by more than 0.3 mg/dL and is diagnosed by kidney biopsy . On pathologic examination , the interstitium of the allograft is diffusely edematous and infiltrated by P01730 and CD8 lymphocytes . Tubulitis occurs when the lymphocytes and monocytes extend into the walls and lumina of the tubules . Presence of leukocytes determines infection or antibody-mediated rejection . Typically C4d staining is negative . Other causes of acute allograft dysfunction included prerenal factors , interstitial nephritis , infection , acute tubular necrosis , toxicity by drugs , and obstruction in the urinary tract . The primary diagnostic assessments include history , especially adherence to immunosuppressive therapy , physical examination , blood and urine laboratory tests , measurement of the serum levels of the drugs , and ultrasonography . Diagnosis of acute cellular rejection depends on biopsy , P11836 staining for refractory cases , negative C4d staining , presence of markers of activating lymphocyte , and proteomic study . Treatment of acute cellular rejection in kidney transplant recipients include pulse steroid for the first rejection episode . It can be repeated for recurrent or resistant rejection . DB00098 and OKT3 are used as the second line of treatment if graft function is deteriorating . Changing the protocol from cyclosporine to tacrolimus or adding mycophenolate mofetil or sirolimus might be effective . Prognosis depends on number of rejection episodes , the use of potent drugs , time of rejection from transplantation , and response to treatment . Redistribution of membrane glycoproteins in platelets activated under flow conditions . A reduction in the ability of GPIb to bind specific MoAbs or ligands ( P04275 ) has been reported in platelets exposed to thrombin in suspension . We have analyzed modifications in the presence of glycoproteins ( GPs ) on platelets activated under flow conditions in a system which allows limited thrombin and fibrin generation . Normal blood anticoagulated with low molecular weight heparin ( LMWH , DB06779 20 IU/ml ) was recirculated for up to 10 min at 800 s-1 through annular chambers containing denuded arterial segments . Aliquots of blood were removed from the reservoir at 0 , 1 , 5 and 10 min and immediately mixed with paraformaldehyde . Membrane glycoproteins : GPIb ( CD42b ) , P08514 -IIIa ( CD41a ) , P16671 ( P16671 ) ; and activation dependent antigens : P16109 ( CD62P ) and lysosomal glycoprortein ( P08962 ) , were detected in whole blood by dual color flow cytometry . Circulation of through the perfusion system resulted in platelet activated as demonstrated by the increased percentage of platelets positive for antigens CD62P and P08962 . A gradual increase in the binding of MoAbs directed against GPIb , P08514 -IIIa , and P16671 epitopes was noted during the entire perfusion period . Observed differences in mean fluorescence intensities at all the observation times were statistically significant ( P < 0.001 ) . Our results obtained on platelets in an experimental thrombosis system indicate that GPIb , P08514 -IIIa and P16671 remain on the surface of activated platelets and actually increase their expression . Alterations detected at the level of GPIb in platelets activated by thrombin in suspension may not take place under in vivo situations . Oocyte-expressed interleukin 7 suppresses granulosa cell apoptosis and promotes oocyte maturation in rats . Development of ovarian follicles is regulated by pituitary-derived gonadotropins together with local ovarian paracrine factors . Based on DNA microarray data , we performed RT-PCR and immunostaining to demonstrate the expression of interleukin 7 transcripts in oocytes of preantral , antral , and preovulatory follicles in rats . We also found the expression of interleukin 7 receptor and the coreceptor interleukin 2 receptor gamma in granulosa cells , cumulus cells , and preovulatory oocytes . In cultured rat granulosa cells obtained from early antral and preovulatory follicles , treatment with interleukin 7 stimulated the phosphorylation of AKT , glycogen synthase kinase ( P49841 ) , and P42229 proteins in a time- and dose-dependent manner . Furthermore , measurement of mitochondrial reductase activity indicated that treatment with interleukin 7 , similar to gonadotropins , increased the number of viable granulosa cells during a 24-h culture period . Furthermore , monitoring of the activities of apoptotic enzymes ( caspase 3/7 ) indicated that treatment with interleukin 7 suppressed apoptosis of cultured granulosa cells from both antral and preovulatory follicles following serum withdrawal . The apoptosis-suppressing actions of interleukin 7 were blocked by an inhibitor of the phosphoinositol-3-kinase ( PIK3 ) /AKT pathway . Furthermore , treatment of cultured preovulatory follicles with interleukin 7 , like treatment with human chorionic gonadotropin , induced germinal vesicle breakdown of oocytes . The stimulatory effect of interleukin 7 was also blocked by inhibitors of the PIK3/AKT pathway . The present findings suggest that oocyte-derived interleukin 7 could act on neighboring granulosa cells as a survival factor and promote the nuclear maturation of preovulatory oocytes through activation of the PIK3/AKT pathway .

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DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . Effectiveness of infliximab , adalimumab and DB06674 for non-infectious refractory uveitis in adults . AIM : To discuss the available data regarding the off-label uses of anti- P01375 agents in non-infectious uveitis . DATA SOURCE : A literature search was performed in Medline through PubMed from January 2001 to January 2014 . STUDY SELECTION AND DATA EXTRACTION : English-language articles about uveitis treatment with anti- P01375 drugs in adult patients were reviewed . DATA SYNTHESIS : The use of anti- P01375 -Î ± drugs for treatment of several refractory manifestations of refractory uveitis in adult patients is increasing . However , due to the lack of evidence from randomized controlled trials , the use of anti- P01375 in uveitis remains “off-label” in most countries . There is no trial-based evidence to support it except for the experience provided by cases and case series . This experience , which is continuously increasing , has yielded encouraging results . Anti- P01375 -Î ± drugs , such as infliximab , adalimumab , and DB06674 , are reasonably effective for controlling ocular inflammation and sparing patients corticosteroid treatment in non-infectious refractory uveitis . Approximately 80 % of patients on infliximab , adalimumab , or DB06674 were able to achieve sustained control of inflammation by 6 months . CONCLUSION : Anti- P01375 -Î ± therapy is effective in inducing clinical remission for refractory uveitis , with a relatively low rate of treatment-ending adverse events . However , randomized and controlled trials are required to adequately assess the maintained clinical efficacy and safety profile in the long term of anti- P01375 agents for non-infectious refractory uveitis . Tuberculosis risk in patients treated with non-anti-tumor necrosis factor-α ( P01375 -α ) targeted biologics and recently licensed P01375 -α inhibitors : data from clinical trials and national registries . This review aimed to evaluate the risk of active tuberculosis ( TB ) occurrence in patients with rheumatic disorders receiving non-anti-tumor necrosis factor ( P01375 ) targeted biologics anakinra ( ANK ) , tocilizumab ( TCZ ) , rituximab ( RTX ) , abatacept ( ABA ) , and recently approved anti- P01375 DB06674 ( GOL ) , and certolizumab pegol ( P53007 ) . In recent findings , no cases of active TB were recorded in patients with rheumatoid arthritis ( RA ) and other rheumatic conditions treated with anti- P11836 + RTX and anti- P10747 ABA . No patient receiving anti-interleukin 1 ( IL-1 ) ANK developed active TB , and an increased risk was excluded in a Canadian database . In contrast , 8 active TB cases were observed in 21 trials of patients with RA receiving anti- P05231 TCZ , while no increased TB risk resulted from Japanese postmarketing surveillance . Among GOL-treated and P53007 -treated patients , 8 and 10 active TB cases occurred , respectively , while no data are available from registries . However , all but 1 TB case recorded in patients treated with TCZ , GOL , and P53007 occurred in TB-endemic countries . No TB risk resulted for ANK , RTX , and ABA , suggesting pretreatment screening procedures for latent TB infection detection are unnecessary . Because all TB cases occurred in countries at high risk for TB , where TB exposure could have occurred during treatment , no definitive conclusions can be drawn for TCZ , GOL , and P53007 . Combination systemic therapies in psoriatic arthritis . Psoriatic arthritis ( PsA ) is a chronic , progressive , and debilitating disorder . When monotherapy fails , combination therapy is necessary for the long-term management of these patients . There is currently no review on this subject , and the purpose of this study was to investigate and describe the current literature on combination therapy in PsA . A PubMed MeSH search was performed for psoriatic arthritis and combination therapy , which yielded at total of 83 articles . After excluding reviews and commentaries , and pursuing relevant citations , a total of 21 articles on the subject were found : one study of NSAIDs and methotrexate , three studies of cyclosporine and methotrexate , three studies of non- P01375 biologic inhibitors ( alefacept , ustekinumab ) and methotrexate , and 14 studies of anti- P01375 -inhibitors ( etanercept , adalimumab , infliximab , DB06674 ) and methotrexate . The combination of cyclosporine and methotrexate reduces the dosages and also the side effects of each agent , allowing better disease control with less toxicity . DB00563 in combination with biologic agents , either non- P01375 inhibitors or anti- P01375 inhibitors , may have a role in decreasing side effects , but it does not appear to improve clinical symptoms beyond those attained by biologic monotherapy . Elevation of P15941 serum levels in clinical trials of tumor necrosis factor inhibitors in patients with rheumatoid arthritis : a report from the Japan College of Rheumatology Ad Hoc Committee for Safety of Biological DMARDs . OBJECTIVE : The associations between elevated levels of serum P15941 ( P15941 ) and treatment of rheumatoid arthritis ( RA ) with tumor necrosis factor ( P01375 ) inhibitors were investigated in five Japanese clinical trials . METHODS : Percentages and incidence rates were calculated for elevated serum P15941 levels . Adverse events associated with elevated levels of serum P15941 were investigated . RESULTS : In RISING , a clinical trial for infliximab , 15.6 % of the enrolled patients met criterion B ( P15941 ≥500 U/ml and > 1.5-fold increase over the baseline value ) by week 54 . In HIKARI , 7.8 % of the certolizumab pegol ( CZP ) group and 0 % of the placebo group met criterion B during the double-blind ( DB ) period ( p = 0.003 ) . In J-RAPID , 8.4 % of the methotrexate ( MTX ) + CZP and 3.9 % of the MTX + placebo groups met criterion B during the DB period . In GO-MONO , 1.8 % of the DB06674 ( GLM ) and 1.3 % of the placebo groups met criterion B during the DB period . In GO-FORTH , 7.1 % of the MTX + GLM and 0 % of the MTX + placebo groups met criteron B during the DB period ( p = 0.017 ) . No adverse events accompanied the elevation of serum P15941 levels in 95.7 % of these patients . CONCLUSION : Serum P15941 levels may increase during anti- P01375 therapy without significant clinical events . In these patients , continuing treatment with P01375 inhibitors under careful observation is a reasonable option . [ New biologic and non biologic disease modifying anti-rheumatic drugs for rheumatoid arthritis ] . P01375 inhibitors , infliximab , etanercept and adalimumab , and tocilizumab are available in Japan and have been successful at improving the signs and symptoms of RA and , thereby , have set a new standard for disease control of RA and have the potential to protect joints from structural damage or to improve quality of life and mortality . However , the rate of successful induction of remission was about 30 % and the treatment strategies for the patients who do not respond to these biologics should be established . Randomized clinical trials targeting T or B lymphocytes have been conducted in addition to the new anti- P01375 blockers like DB06674 or certolizumab pegol . Anti- P11836 antibodies such as rituximab ( chimeric ) , ocrelizumab ( humanized ) , ofatuzumab ( full human ) demonstrated effectiveness to the patients who do not respond to P01375 blockers . P16410 Ig , which can transduce negative signal into T lymphocytes in the co-stimulatory pathway , has also showed a good response to refractory RA . Furthermore , low molecular agents such as Jak ( Janus kinase ) 3 or syk ( spleen tyrosine kinase ) inhibitors demonstrated rapid and strong suppression of synovitis and are thought to be new candidates for the drugs to overcome refractory RA . Biologic agents for rheumatoid arthritis : 2008 and beyond . Rheumatoid arthritis ( RA ) is a chronic disease with a complex underlying pathology and varied presentation in patients . Several novel biologic disease-modifying antirheumatic drugs have become available for the treatment of RA . Agents in late-stage clinical trials include DB06674 and certolizumab , which are anti-tumor necrosis factor ( P01375 ) -alpha agents ; ocrelizumab , an anti- P11836 agent ; and tocilizumab , an inhibitor of interleukin-6 . As treatment options for RA expand , nursing care will play an increasingly important role in empowering patients through interventions such as patient education and adverse effect management . Treatment of psoriasis and psoriatic arthritis . Skin and joint manifestations associated with psoriasis and psoriatic arthritis ( PsA ) can significantly impact a patient 's quality of life . Successful treatment is imperative in order to improve signs and symptoms of the disease , and to alleviate physical or psychological distress . For patients with mild psoriasis with or without PsA , topical agents and targeted phototherapy are appropriate treatments for psoriasis . Systemic therapies , such as methotrexate and phototherapy are recommended options for patients with more severe psoriasis , but their long-term use is hindered by safety concerns . Advancements in understanding the pathogenesis of psoriasis , including the role of T cells and cytokines , have been crucial to the development of biological therapies . These target the immune system and are suitable options for patients with extensive disease . Biological therapies for the treatment of psoriasis include targeted therapies ( alefacept ) and anti-cytokine therapies ( anti-tumour necrosis factor [ P01375 ] therapies [ adalimumab , etanercept , infliximab ] and a monoclonal antibody against interleukin [ IL ] -12 and IL-23 [ ustekinumab ] ) . Patients with PsA should be treated appropriately in order to improve symptoms and inhibit structural joint damage . Non-steroidal anti-inflammatory drugs or local intra-articular injections of corticosteroids can be used successfully in patients with mild PsA ; however , neither treatment prevents the development of structural joint damage . For patients with moderate to severely active PsA , disease-modifying antirheumatic drugs ( such as methotrexate ) , P01375 inhibitor treatments ( adalimumab , etanercept , infliximab and DB06674 ) or their combination are considered first-line treatment . This review provides a brief overview of treatment options for psoriasis and PsA , with an emphasis on the efficacy and safety of anti- P01375 therapies . Immunogenicity of anti- P01375 biologic therapies for rheumatoid arthritis . Currently , five anti- P01375 biologic agents are approved for the treatment of rheumatoid arthritis ( RA ) : adalimumab , infliximab , etanercept , DB06674 and certolizumab pegol . Formation of anti-drug antibodies ( P00813 ) has been associated with all five agents . In the case of adalimumab and infliximab , immunogenicity is strongly linked to subtherapeutic serum drug levels and a lack of clinical response , but for the other three agents , data on immunogenicity are scarce , suggesting that further research would be valuable . Low P00813 levels might not influence the efficacy of anti- P01375 therapy , whereas high P00813 levels impair treatment efficacy by considerably reducing unbound drug levels . Immunogenicity is not only an issue in patients treated with anti- P01375 biologic agents ; the immunogenicity of other therapeutic proteins , such as factor VIII and interferons , is well known and has been investigated for many years . The results of such studies suggest that investigations to determine the optimal treatment regimen ( drug dosing , treatment schedule and co-medication ) required to minimize the likelihood of P00813 formation might be an effective and practical way to deal with the immunogenicity of anti- P01375 biologic agents for RA . Golimumab and malignancies : true or false association ? Malignancy is one of the comorbidities linked to DB06674 , a biological P01375 -α blocker . In this systematic review and meta-analysis , we searched different databases and analyzed original publications to elucidate the remaining open question about the real association of malignancies with DB06674 therapy . The most frequent cancer in patients treated with DB06674 , in association or not with methotrexate , is the lung adenocarcinoma . However , lymphoma is not very commonly represented in these patients . We show that there is no major and evident risk of malignancies associated with DB06674 in current scientific literature . An increased risk of malignancies may be associated with DB06674 , but this warrants further clinical confirmation . Also , this risk mentioned in different studies must be taken with caution because of number of limits and biases . Regulation of P14061 and P18405 in lymphocytes . We previously reported lymphocyte expression of genes encoding enzymes required for steroid metabolism ; however , only 17beta-HSD and 5alpha-reductase showed significant enzyme activity . We now investigate regulation of lymphocyte expression for genes encoding 17beta-HSD and 5alpha-reductase . Cultured human T and B lymphoid cell lines and peripheral blood mononuclear cells were treated with known regulators of steroidogenic gene expression including forskolin , PMA , ionomycin , various steroids , interleukin ( IL ) -4 , and P05231 . Treatment with 10 or 50 microM forskolin resulted in a 20-60 % reduction of expression for P14061 ( encoding 17beta-HSD I ) in T and B lymphoid cell lines and peripheral blood mononuclear cells , although such a change was not observed in the expression of P18405 ( encoding 5alpha-reductase I ) . No significant changes were found when cells were treated for 24 h with various concentrations of PMA or ionomycin . Incubation with 10(-9) to 10(-7) M androstenedione or estradiol increased expression of P14061 , while testosterone decreased the expression of this gene . P18405 expression was increased in the presence of 5alpha- DB02901 although no consistent changes were observed when the cells were treated with testosterone . Other steroids , including dexamethasone , progesterone , and 6-hydroxypregnanolone , produced no effects on expression of either P14061 or P18405 . Treatment with 0.1-10 ng/ml of P05112 or P05231 also did not effect significant changes in gene expression . These data implicate the involvement of the DB02527 -protein kinase signal transduction pathway in regulating lymphocyte expression of P14061 . Furthermore , it appears that lymphocyte P14061 and P18405 are regulated to some extent by specific steroids . Tumor necrosis factors blocking agents : analogies and differences . Five anti- P01375 agents , infliximab , adalimumab , etanercept , DB06674 and certolizumab pegol are approved worldwide for the treatment of RA . Anti- P01375 agents , bind to and neutralize soluble P01375 , but exert different effects on transmembrane P01375 -expressing cells ( P01375 -producing cells ) . Differences on affinity and avidity for soluble and transmembrane P01375 were showed . Different activity on cells apoptosis , complement-dependent cytotoxicity ( CDC ) antibody dependent cell-mediated cytotoxicity ( Q15848 ) were described . Some dramatic changes in gene expression were seen with all the anti-TNFs . Reviewing the biology of transmembrane P01375 and its interaction with anti- P01375 agents will contribute to understanding the bases of differential clinical efficacy of these promising treatment modalities . Targeting tumor necrosis factor alpha in psoriasis and psoriatic arthritis . BACKGROUND : Psoriasis is an immune-mediated chronic inflammatory disease triggered and maintained by inflammatory mediators , including P01375 . OBJECTIVE/METHODS : To summarize the role of anti- P01375 agents psoriasis therapy , focusing on the mechanisms and biological pathways involved , by reviewing relevant literature . RESULTS/CONCLUSIONS : The three P01375 antagonists currently available ( etanercept , infliximab and adalimumab ) are effective in the therapy of psoriasis and psoriatic arthritis . DB08904 and DB06674 are P01375 inhibitors not approved for therapy of psoriasis yet . In addition to neutralizing soluble P01375 , P01375 blockers bind to membrane P01375 and change the behavior of P01375 -expressing cells , resulting in hastened cell cycle arrest and apoptosis , and suppression of cytokine production . P01375 blockers may also affect adaptive immune responses by reducing T helper cell (Th)1 and Th17 responses , and favoring the development of T-regulatory cells . P01375 antagonists can regulate differentiation and activation of osteoclasts , thus reducing bone destruction in psoriatic arthritis . Anti- P01375 agents differ in their pharmacokinetics and pharmacodinamic properties , which is reflected in their therapeutic and safety profiles . The safety of P01375 antagonists has been established , and patient selection and monitoring allow risk minimization . P01375 inhibitors in asthma and P48444 : we must not throw the baby out with the bath water . P01375 ( P01375 ) -alpha , a pleiotropic cytokine that exerts a variety of effects , such as growth promotion , growth inhibition , angiogenesis , cytotoxicity , inflammation , and immunomodulation , has been implicated in several inflammatory conditions . It plays a significant role in many inflammatory diseases of lungs . Given that there is significant literature supporting the pathobiologic role of P01375 in asthma , mainly in severe refractory asthma , and P48444 , P01375 inhibitors ( infliximab , DB06674 and etanercept ) are now regarded as the potential new medications in asthma and P48444 management . The studies reported in literature indicate that P01375 inhibitors are effective in a relatively small subgroup of patients with severe asthma , possibly defined by an increased P01375 axis , but they seem to be ineffective in P48444 , although an observational study demonstrated that P01375 inhibitors were associated with a reduction in the rate of P48444 hospitalisation among patients with P48444 receiving these agents to treat their rheumatoid arthritis . These findings require a smart approach because there is still good reason to target P01375 , perhaps in a more carefully selected patient group . P01375 treatment should , therefore , not be thrown out , or abandoned . Indeed , since severe asthma and P48444 are heterogeneous diseases that have characteristics that occur with different phenotypes that remained poorly characterized and little known about the underlying pathobiology contributing to them , it is likely that definition of these phenotypes and choice of the right outcome measure will allow us to understand which kind of patients can benefit from P01375 inhibitors . Q9UKU7 : Golimumab in ulcerative colitis : a ' ménage à trois ' of drugs . Golimumab , a human anti- P01375 antibody , is effective in patients with ulcerative colitis , according to new findings from an international phase III double-blind trial . The addition of this drug makes a ménage à trois of available drugs -- comprising infliximab , adalimumab and DB06674 -- for the treatment of ulcerative colitis . Anti- P01375 agents in familial Mediterranean fever : report of three cases and review of the literature . Familial Mediterranean fever ( FMF ) is an autoinflammatory disease characterized by recurrent fever , peritonitis/pleuritis , or arthritis attacks . Patients may have FMF-associated mutations of pyrin . The role of biologics such as anti-tumor necrosis factor ( P01375 ) agents ( infliximab , etanercept , adalimumab , DB06674 ) and anakinra , canakinumab , or rilonacept in the treatment of FMF needs to be clarified . Herein we present reports of three patients ( all were positive for HLA Q8TCY5 ) with typical spondylitis associated with FMF who were successfully managed with anti- P01375 agents , along with a literature review . The patients were a 37-year-old man with concomitant Crohn 's disease and amyloidosis who was treated with infliximab ( P27352 , 5 mg/kg for 3 years ) and switched to adalimumab ( P00813 ) , and two female patients ( a 24-year-old and a 31-year-old ) with FMF who developed severe spondylitis and who were also treated with P00813 . Anti- P01375 agents can control FMF attacks quite effectively and they reveal a promising role in the treatment of FMF-associated amyloidosis and spondylitis . Efficacy and safety of P01375 -α inhibitors in refractory primary complex aphthosis : a patient series and overview of the literature . BACKGROUND : Otherwise healthy patients with severe recurrent mucocutaneous aphthous ulcerations ( complex aphthosis ) may require systemic immunomodulatory therapy . However , a subset of patients remain resistant or intolerant to recommended therapeutic agents . Recently , case reports have described that tumor necrosis factor-α ( P01375 -α ) inhibitors may induce remission in these patients . METHODS : Data on efficacy and safety of various P01375 -α inhibitors used as monotherapy in a case series of 18 patients with refractory primary complex aphthosis are presented . RESULTS : A total of 16 patients ( 89 % ) obtained complete or almost clearance of orogenital aphthous ulcerations rapidly after onset of therapy either with etanercept , adalimumab , infliximab or DB06674 . Duration of treatment ranged between 3 and 77 months . Nine patients ( 50 % ) received more than one P01375 -α inhibitor during the course of treatment . Five ( 28 % ) patients experienced side effects that could be related to treatment with P01375 -α inhibitors . CONCLUSION : P01375 -α inhibitors are an effective and safe treatment option for patients with severe complex aphthosis who do not respond sufficiently to standard therapy as recommended by existing guidelines . However , the final position of P01375 -α inhibitors in the therapeutic armamentarium awaits randomized controlled trials . Off-label uses of anti- P01375 therapy in three frequent disorders : Behçet 's disease , sarcoidosis , and noninfectious uveitis . Tumoral necrosis factor α plays a central role in both the inflammatory response and that of the immune system . Thus , its blockade with the so-called anti- P01375 agents ( infliximab , etanercept , adalimumab , certolizumab pegol , and DB06674 ) has turned into the most important tool in the management of a variety of disorders , such as rheumatoid arthritis , spondyloarthropatties , inflammatory bowel disease , and psoriasis . Nonetheless , theoretically , some other autoimmune disorders may benefit from these agents . Our aim is to review these off-label uses of anti- P01375 blockers in three common conditions : Behçet 's disease , sarcoidosis , and noninfectious uveitis . Due to the insufficient number of adequate clinical trials and consequently to their lower prevalence compared to other immune disorders , this review is mainly based on case reports and case series . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . P01375 inhibitors in psoriasis : an update . Three inhibitors of tumor necrosis factor ( P01375 ) currently are approved for the treatment of psoriasis : etanercept , infliximab , and adalimumab . The other two P01375 inhibitors , DB06674 and certolizumab pegol , have shown efficacy against plaque psoriasis in clinical trials of psoriatic arthritis ( PsA ) . This article reviews the most recent evidence on the efficacy and safety of the P01375 inhibitors in psoriasis , with special attention to preventing and managing immunogenicity . Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin-1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA/uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 ) or interleukin-1 beta ( P01584 ) , are present in epidermal wounds . We have therefore tested P01584 and P01375 for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 and P01375 induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing . Current , new and future biological agents on the horizon for the treatment of inflammatory bowel diseases . Biological agents for inflammatory bowel diseases ( Q9UKU7 ) targeting tumor necrosis factor ( P01375 ) have changed the way to treat Q9UKU7 refractory to standard medications and allowed us to reach new therapeutic goals such as mucosal healing and deep remission . A better understanding of the components of the pathological processes that are a hallmark of Q9UKU7 has led to the development of a new family of biological agents in Crohn 's disease and ulcerative colitis . Biosimilars , which are copy versions of currently licensed biological agents , will be soon available . The biosimilar of infliximab is as effective and as safe as its originator in rheumatologic conditions , while a new anti- P01375 agent , namely DB06674 , has been recently approved for refractory ulcerative colitis . Beyond P01375 blockers , anti-adhesion molecules appear to be a potent drug class for Q9UKU7 . DB09033 was recently approved for both Crohn 's disease and ulcerative colitis . Numerous other compounds are in the pipeline . Ustekinumab looks very promising for Crohn 's disease . O15105 antisense oligonucleotide might enrich our armamentarium if preliminary data are confirmed in upcoming clinical trials . Herein , we review the efficacy and safety of new and emerging biological agents that are currently investigated in Q9UKU7 clinical trials . Biologic therapies in the treatment of psoriasis : a comprehensive evidence-based basic science and clinical review and a practical guide to tuberculosis monitoring . The treatment of psoriasis has undergone a revolution with the advent of biologic therapies including infliximab , etanercept , adalimumab , efalizumab , DB06674 , certolizumab , alefacept , secukinumab , abatacept , and ustekinumab . These medications are designed to target specific components of the immune system and are a major technological advancement over traditional immunosuppressive medications . Herein , we present a comprehensive , unbiased comparison of these medications focusing on their differences . For example , P01375 antagonists can differ in the way they are dissolved and administered , the effector molecules they can bind , serum peak and trough levels , the types of intracellular signals they can induce , the in vivo complexes that they can form , their protein structure , and their incidence and timing of rare adverse events , among other things . A critical review of the clinical studies that have tested the efficacy of these molecules is also presented including head-to-head comparison trials . The safety of biologics in terms of their long-term adverse events is discussed , as is their use in different types of psoriasis and in different patient populations . Finally , all anti- P01375 agents have been associated with a variety of serious and " routine " opportunistic infections , particularly tuberculosis . For this reason , anti-tuberculosis testing both prior to the initiation of a biologic therapy and annually during treatment is pertinent . The uses and limitations of both the tuberculin skin test ( Q16762 ) and QuantiFeron®-TB Gold ( QFT ) are discussed , as is the care of patients who present with latent tuberculosis infection prior to the initiation of biologic therapy . Recommendations for tuberculosis monitoring are provided . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . Golimumab and immunogenicity ? 2010 and beyond . Immunogenicity is a frequent adverse event observed with biological agents ' therapy . Challenges of management in patients with rheumatoid arthritis , psoriatic arthritis and ankylosing spondylitis treated with DB06674 , an anti- P01375 blocker , include limited generation of antibodies like anti-nuclear , anti- DB06674 , and anti-double stranded DNA antibodies . We conducted here a meta-analysis study in order to evaluate and compare the newly generated antibody levels after DB06674 therapy . The examination of original clinical trials revealed that their levels were neither higher nor significant . Moreover , no evident associations between the induced-antibodies and lupus-like syndromes and/or infusion site reaction were reported . The reduced patients cohort and the absence of systematic newly generated antibodies follow-up might be implicated in the difficulty to evaluate their risk in delaying diseases therapy , and/or predicting for their worse prognosis . Hence , further studies are required to ascertain the real impact of the induced antibodies after DB06674 's therapy . Safety of anti- P01375 therapy in inflammatory bowel disease during pregnancy . INTRODUCTION : The highest incidence of inflammatory bowel disease ( Q9UKU7 ) is seen between the second and fourth decades of life , which is the most fertile age for women . Increased disease activity has been shown to effect female fertility and pregnancy outcomes , stressing the need for drugs that can safely induce and maintain clinical remission without harming either the mother or fetus . AREAS COVERED : Anti- P01375 -α agents have been shown to be effective in both inducing and maintaining remission among Q9UKU7 patients . This review highlights the results of previous studies conducted on pregnant women who were exposed to anti- P01375 -α agents during the course of their pregnancy . The drugs reviewed include infliximab ( IFX ) , adalimumab ( P00813 ) , certolizumab pegol ( CZP ) and DB06674 ( GMB ) . Of > 200 articles reviewed , 105 were included in the manuscript based on relevance . The keywords used were anti- P01375 , infliximab , adalimumab , certolizumab , DB06674 , biologics , pregnancy and inflammatory bowel disease . EXPERT OPINION : Anti- P01375 agents have been studied extensively during pregnancy from the early case reports to the more recent prospective Pregnancy in Q9UKU7 and Neonatal Outcomes study . A comprehensive review of the literature has shown that biologics can be safely used during pregnancy . In view of this safety data , it is recommended to maintain therapy during pregnancy . Synergism between bosutinib ( DB06616 ) and the Chk1 inhibitor ( PF-00477736 ) in highly imatinib-resistant P11274 /ABL⁺ leukemia cells . Interactions between the dual P11274 / P00519 and Src inhibitor bosutinib and the Chk1 inhibitor PF-00477736 were examined in P11274 / P00519 (+) leukemia cells , particularly imatinib-resistant cells , including those with the T315I mutation . Bosutinib blocked PF-00477736-induced P27361 /2 activation and sharply increased apoptosis in association with Mcl-1 inhibition , p34(cdc2) dephosphorylation , BimEL up-regulation , and DNA damage in imatinib-resistant CML or Ph(+) ALL cell lines . Inhibition of Src or Q02750 by shRNA significantly enhanced PF-0047736 lethality . Bosutinib/PF-00477736 co-treatment also potentiated cell death in P28906 (+) CML patient samples , including dasatinib-resistant blast crisis cells exhibiting both T315I and E355G mutations , but was minimally toxic to normal P28906 (+) cells . Finally , combined in vivo treatment significantly suppressed BaF3/T315I tumor growth and prolonged survival in an allogeneic mouse model . Together , these findings suggest that this targeted combination strategy warrants attention in IM-resistant CML or Ph(+) ALL . Safety and clinical efficacy of DB06674 in the treatment of arthritides . Golimumab is a human anti-tumor necrosis factor ( P01375 ) -alpha monoclonal antibody that was recently approved for the treatment of patients with rheumatoid arthritis , psoriatic arthritis , and ankylosing spondylitis . This review covers the published clinical trial data on the use of DB06674 for the approved indications mentioned above with respect to efficacy and safety . The various ongoing trials for DB06674 have yielded promising results in terms of efficacy and safety in methotrexate-naive and -resistant patients with rheumatoid arthritis , as well as in patients who were previously treated with other anti- P01375 agents . In addition , the efficacy of DB06674 in psoriatic arthritis and ankylosing spondylitis has also been demonstrated . The real safety information will be available only once the drug has been used in many more patients , who frequently have comorbid conditions . New therapies in the management of rheumatoid arthritis . PURPOSE OF REVIEW : The therapeutic landscape in the management of rheumatoid arthritis ( RA ) has witnessed significant changes over the past decade . The ambition to improve outcomes further , minimize safety concerns and provide more convenient means of administration are all factors that continue to drive continued drug development . This review summarizes novel therapies that have been most recently under investigation . RECENT FINDINGS : More refined drug technology has seen the development of subcutaneous forms of existing therapies ( abatacept , tocilizumab ) , as well as newer-generation monoclonal antibodies ( e.g. B-cell-depleting agents , ocrelizumab and ofatumumab and the P01375 -inhibitors certolizumab and DB06674 ) . Alternative methods of targeting critical pathways , for example Blys inhibition ( atacicept ) and P05231 as opposed to P05231 receptor antagonism , have also been evaluated . Finally , small molecules are receiving increasing attention , with some of the protein kinases inhibitors particularly promising . SUMMARY : The new emerging therapies for the management of RA illustrate much diversity , in terms of both drug technology as well as the immunological target . Although not all may succeed in reaching the market , important insights can still be gained . Challenging and exciting times lie ahead as these new technologies are embraced and efforts are made to determine how best to implement in practice . Round window membrane permeability to DB06674 in guinea pigs : a pilot study . OBJECTIVES/HYPOTHESIS : Autoimmune inner ear disorder is one of a few types of sensorineural hearing loss that is treatable and potentially reversible . Treatment involves oral steroids and methotrexate . Other treatment modalities have been tried with variable success . All such treatments are systemic , with inherent side effects limiting their effectiveness . Recently , tumor necrosis factor ( P01375 ) -α blockers have been suggested as a modality of treatment . The objective of this study was to assess the round window membrane permeability to DB06674 , a P01375 -α blocker . This study is the first to look at the feasibility of local DB06674 delivery into the inner ear , which may allow for targeted immune modulation of autoimmune inner ear disorders without the consequences of systemic treatment . STUDY DESIGN : This is a single-blinded , placebo-controlled , pilot study using guinea pigs to assess round window membrane permeability to DB06674 . METHODS : Golimumab was instilled into the guinea pigs ' middle ear . Inner ear fluid was sampled through the round window membrane after approximately 30 minutes of drug exposure . Golimumab presence in the inner ear was assessed by enzyme-linked immunosorbent assay in both drug-treated and control ears . RESULTS : Higher concentrations of DB06674 were detected in the inner ear fluid samples of DB06674 -exposed ears than in the control ears . The difference was statistically significant ( P < .001 ) . CONCLUSIONS : Golimumab crosses the round window membrane and is detected in measurable concentrations in the inner ear fluid after 30 minutes of exposure to the membrane . Further studies are needed to learn its pharmacokinetics and the time needed to reach optimal concentration in the inner ear . LEVEL OF EVIDENCE : NA. Laryngoscope , 123:2840-2844 , 2013 . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . Dermatologic adverse events : DB06674 , friend or foe ? Golimumab is a fully human anti- P01375 blocker that has demonstrated its efficacy in the treatment of numerous kinds of diseases . Although it is generally safe and well tolerated , various adverse events have been reported . The present aim is to improve the understanding of dermatologic adverse events associated with DB06674 following a search of various scientific databases . This systematic review and meta-analysis shows that DB06674 is associated neither with severe injection-site reactions nor with injection-site erythema . We found no significant lupus-like syndromes , and no significant skin squamous cell carcinoma . We further suggest systematic dermatologic monitoring in clinical practice during DB06674 therapy . Subsequent research should employ a larger cohort of patients to ensure clear and significant future conclusions . DB00877 unbalances the polarization of human macrophages to M1 . Plasticity is a hallmark of macrophages , and in response to environmental signals these cells undergo different forms of polarized activation , the extremes of which are called classic ( M1 ) and alternative ( M2 ) . DB00877 ( Q96PN7 ) is crucial for survival and functions of myeloid phagocytes , but its effects on macrophage polarization are not yet studied . To address this issue , human macrophages obtained from six normal blood donors were polarized to M1 or M2 in vitro by lipopolysaccharide plus interferon-γ or interleukin-4 ( P05112 ) , respectively . The presence of Q96PN7 ( 10 ng/ml ) induced macrophage apoptosis in M2 but not in M1 . Beyond the impact on survival in M2 , Q96PN7 reduced P61073 , CD206 and Q9NNX6 expression and stem cell growth factor-β , P55774 and Q99616 release . In contrast , in M1 Q96PN7 increased P42081 and P32248 expression and P05231 , tumour necrosis factor-α and IL-1β release but reduced CD206 and Q9NNX6 expression and P22301 , vascular endothelial growth factor and P55774 release . In view of the in vitro data , we examined the in vivo effect of Q96PN7 monotherapy ( 0·1 mg/kg/day ) in 12 patients who were treated for at least 1 month before islet transplant . Cytokine release by O00206 -stimulated peripheral blood mononuclear cells showed a clear shift to an M1-like profile . Moreover , macrophage polarization 21 days after treatment showed a significant quantitative shift to M1 . These results suggest a role of mammalian target of rapamycin ( P42345 ) into the molecular mechanisms of macrophage polarization and propose new therapeutic strategies for human M2-related diseases through P42345 inhibitor treatment . Two cases of Takayasu 's arteritis occurring under anti- P01375 therapy . Takayasu 's arteritis is a granulomatous , large vessel vasculitis that affects the aorta , its major branches and the pulmonary arteries . Compelling evidence exists to support the notion that Takayasu 's arteritis is a T-cell mediated process and that tumor necrosis factor alpha ( TNFa ) is an important factor in the pathogenesis of this disease . Moreover , encouraging results from recent studies support the use of anti-TNFa therapy for relapsing or resistant cases of Takayasu 's arteritis . Here , however , we describe the case of two patients : one with seropositive rheumatoid arthritis , the other with HLA- Q8TCY5 negative spondylarthropathy , who developed Takayasu 's arteritis during treatment with TNFa inhibitors ( adalimumab and DB06674 respectively ) . This is the first report of Takayasu 's arteritis in rheumatic patients under TNFa blocking agents which suggests the presence of different pathogenetic mechanism in a subgroup of patients with Takayasu 's arteritis , as well as a potential role of TNFa blockers as triggers of this disease in some cases . Miscellaneous adverse events with biologic agents ( excludes infection and malignancy ) . Anti-tumor necrosis factor-α ( anti- P01375 ) agents are frequently used in the treatment of inflammatory bowel disease ( Q9UKU7 ) . Currently , there are 4 anti- P01375 therapies that are Food and Drug Administration-approved for moderate to severe Q9UKU7 : infliximab , adalimumab , DB06674 , and certolizumab pegol . For most noninfectious , nonmalignant adverse events , cessation of anti- P01375 therapy typically leads to improvement or resolution of drug-induced complications . In this article , the current knowledge regarding the noninfectious and nonmalignant toxicities associated with anti- P01375 agents is summarized . Psoriatic arthritis : treatment strategies using biologic agents . The traditional management of psoriatic arthritis ( PsA ) includes NSAIDs , corticosteroids and DMARDs . Advancement in the knowledge of the immunopathogenesis of PsA has been associated with the development of biologic agents which have revolutionized the management of the disease . Among biologics drugs , there are the 4 currently available anti-TNFα blocking agents ( etanercept , infliximab , adalimumab and DB06674 ) which are more effective than traditional DMARDs on symptoms/signs of inflammation , quality of life , function , and in inhibiting the progression of the structural joint damage . Despite of the high cost , P01375 inhibitors are cost-effective on both the musculoskeletal and skin manifestations of psoriatic disease . Golimumab : clinical update on its use for ulcerative colitis . Monoclonal antibodies directed against tumor necrosis factor alpha ( anti- P01375 -α agents ) have dramatically changed the therapeutical approach to inflammatory bowel diseases , such as Crohn 's disease and ulcerative colitis . A new anti- P01375 drug , DB06674 , has recently been approved for patients with moderate to severe ulcerative colitis . Its efficacy has been demonstrated by preclinical and clinical studies and the drug showed an efficacy and safety profile in line with the other anti- P01375 agents , such as infliximab and adalimumab . This review gives an overview on DB06674 in the treatment of moderate to severe ulcerative colitis . P01375 -alpha antagonists : differential clinical effects by different biotechnological molecules . Inhibitors of tumor necrosis factor-alpha have deeply changed the therapy of several inflammatory human diseases . For instance , clinical management of rheumatoid arthritis , psoriatic arthritis and ankylosing spondylitis have profoundly benefited after the introduction of new therapeutic tools , such as antagonist of P01375 molecule . These drugs include etanercept , a soluble P01375 receptor antagonist , three anti- P01375 antibodies , adalimumab , infliximab , DB06674 and certolizumab a humanized Fab fragment combined with polyethylene glycol . These compounds efficiently inhibit several P01375 biological-mediated effects , however , they have also shown differential clinical efficacy in several trials from different autoimmune diseases . It is of clinical relevance that non-responders to one of these drugs often positively responded to another . Different mechanisms of action and diversity in pharmacokinetics of these three compounds may partially explain different clinical effects . However , partially diverse pathogenetic mechanisms in different diseases also contribute to differential therapeutic responses . Therefore , these apparently homogeneous agents can not be considered equivalent in their clinically efficacy . Differential therapeutic actions of these drugs may be advantageously used in clinical practice and further improve the great potential of individual P01375 inhibitors . Treatment of ankylosing spondylitis with P01375 blockers : a meta-analysis . Biological agents directed against tumor necrosis factor ( P01375 ) represent therapeutic options for patients with ankylosing spondylitis with high disease activity despite use of non-steroidal anti-inflammatory drugs . To evaluate the efficacy and safety of the anti- P01375 agents infliximab , etanercept , adalimumab , DB06674 , and certolizumab for the treatment of ankylosing spondylitis , we performed a systematic review of randomized clinical trials on adult patients with ankylosing spondylitis using articles culled from the EMBASE , MEDLINE , Cochrane Controlled Trials Register and LILACS databases ( September/2012 ) , manual literature search , and the gray literature . Study selections and data collection were performed by two independent reviewers , with disagreements solved by a third reviewer . The following outcomes were evaluated : ASAS 20 response , disease activity , physical function , vertebral mobility , adverse events , and withdraws . The meta-analysis was performed using the Review Manager(®) 5.1 software by applying the random effects model . Eighteen studies were included in this review . No study of certolizumab was included . Patients treated with anti- P01375 agents were more likely to display an ASAS 20 response after 12/14 weeks ( RR 2.21 ; 95 % CI 1.91 ; 2.56 ) and 24 weeks ( RR 2.68 ; 95 % CI 2.06 ; 3.48 ) compared with controls , which was also true for several other efficacy outcomes . Meta-analysis of safety outcomes and withdraws did not indicate statistically significant differences between treatment and control groups after 12 or 30 weeks . DB00051 , infliximab , etanercept , and DB06674 can effectively reduce the signs and symptoms of the axial component of ankylosing spondylitis . Safety outcomes deserve further study , especially with respect to long-term follow-ups . Selective P01375 -α inhibitor-induced injection site reactions . INTRODUCTION : During the last decade , many new biological immune modulators entered the market as new therapeutic principles . P01375 -α is a pro-inflammatory cytokine known to a have a key role in the pathogenic mechanisms of various immune-mediated or inflammatory diseases . P01375 -α blockers have demonstrated efficacy in large , randomized controlled clinical trials either as monotherapy or in combination with other anti-inflammatory or disease-modifying anti-rheumatic drugs . AREAS COVERED : Although generally well tolerated and safe , potential adverse events may be associated with P01375 -α inhibitor treatment . The authors will briefly review the potential adverse drug reactions and the immunological mechanisms of injection site reactions ( ISRs ) in patients treated with etanercept and adalimumab . EXPERT OPINION : Patients treated with P01375 -α inhibitors can develop ISR around the sites of injections . ' Type IV delayed type reaction ' or ' recall ISRs ' . Eosinophilic cellulitis or ' Wells syndrome ' , ' type III ' and ' type I ' reactions are reported . Long-term studies are necessary to determine the durability of response and the real risk of ISRs with DB06674 and certolizumab pegol . Further studies are also necessary to evaluate the immunogenicity of these drugs . Golimumab in uveitis previously treated with other anti- P01375 drugs : a retrospective study of three cases from a single centre and literature review . OBJECTIVES : The aim of this paper is to assess the clinical response to DB06674 ( GLM ) in patients with non-infectious uveitis from a single centre that had previously been treated with other anti- P01375 -α drugs . METHODS : A retrospective chart review was carried out of patients with immune-mediated uveitis refractory to standard synthetic immunosuppressive drugs who were treated with GLM at Hospital Universitario Marqués de Valdecilla , Santander ( Spain ) . Patients were included in this study if they had previously been treated with other anti- P01375 -α drugs . A literature review of patients with immune-mediated uveitis undergoing GLM therapy was conducted . RESULTS : Three patients ( 2 men and 1 woman ) were included in this study . Two of them were refractory to other anti- P01375 -α drugs . The median age of patients was 26 years ( range 20-42 ) . Uveitis was bilateral in two patients . The underlying diseases were uveitis associated with HLA- Q8TCY5 and psoriasis in one case and sarcoidosis in the other two cases . Improvement of the main ocular parameters following GLM therapy was achieved in all cases . After a median follow-up of 3 ( range 1-9 ) months using GLM therapy , none of the patients had experienced new relapses of uveitis . None of them had side effects during treatment with this drug . A literature review disclosed that our observations were in keeping with other reports that showed good response to GLM in 13 of 16 patients with immune-mediated uveitis refractory to other biologic agents . CONCLUSIONS : Although the follow-up was too short in our series , GLM could be an effective and safe therapy for the management of patients with uveitis previously treated with other anti- P01375 -α drugs . NSA9 , a human prothrombin kringle-2-derived peptide , acts as an inhibitor of kringle-2-induced activation in EOC2 microglia . In neurodegenerative diseases , such as Alzheimer 's and Parkinson 's , microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds . P00734 and kringle-2 increase levels of NO and the mRNA expression of P35228 , IL-1beta , and P01375 in microglial cells . In contrast , the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1beta , P01375 , and P05231 in LPS-activated EOC2 microglia . In this study , we investigated the anti-inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia . Treatment with 20-100 muM of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation . NO production induced by MAPKs and NF-kappaB was similarly reduced by inhibitors of P29323 ( PD98059 ) , p38 ( SB203580 ) , NF-kappaB ( DB06151 ) , and NSA9 . These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2 . The use of biologic therapies in the treatment of rheumatoid arthritis . The use of biologic agents has revolutionized the management of rheumatoid arthritis ( RA ) in the past two decades . These biologic agents directly target molecules and cells involved in the pathogenesis of RA . Biologic agents indeed lead to a better prognosis and clinical remission in patients with RA , especially in patients who are not well-controlled with traditional disease-modifying anti-rheumatic drugs ( DMARDs ) . Currently , five P01375 inhibitors ( infliximab , etanercept , adalimumab , DB06674 and certolizumab pegol ) , an P05231 receptor antagonist ( tocilizumab ) , an IL-1 receptor antagonist ( anakinra ) , a B cell depleting agent ( rituximab ) and a T cell co-stimulation inhibitor ( abatacept ) have been approved for the treatment of RA . With the increased understanding of the pathogenic mechanisms of RA and advantages in manufacturing biotechnology of pharmaceutical companies , a series of novel biologic therapeutic approaches are being developed . In the present paper , we will summarize the biologic agents currently available to treat RA , and the prospective biologic therapies that might be used in the management of RA in future . Experimental autoimmune encephalomyelitis in the Wistar rat : dependence of MBP-specific T cell responsiveness on P33681 costimulation . Experimental autoimmune encephalomyelitis ( EAE ) is an animal model of human multiple sclerosis that requires the activation of autoreactive T cells for the expression of pathology . EAE has been most frequently studied in the Lewis rat model as well as in several murine models of EAE including the PLJ and B10PL strains . In the present study we describe a novel model of EAE induced in the Wistar rat strain by immunization with guinea pig spinal cord antigens and pertussis toxin ( PT ) . T cell responses were induced to myelin basic protein . Autoreactive T cells could be totally blocked by the in vitro treatment with DB01281 , a protein that blocks the costimulation of autoreactive T cells . The addition of P60568 could reverse the inhibition seen in vitro with DB01281 . The effects of inhibition of P33681 costimulation were also examined by an analysis of cytokine responses and P60568 receptor on T cells . DB01281 treatment in vitro reduced the expression of P60568 receptor on T cells , enhanced T cell apoptosis and decreased the synthesis of P60568 , P01579 and P01375 . DB01281 treatment had no effect on P22301 synthesis by T cells , a cytokine implicated in the functions of regulatory T cell subsets . Overall , our studies support the rationale of P33681 blocking therapies as a potential treatment for models of multiple sclerosis . The induction of EAE in the Wistar rat provides yet another novel model in which to examine the regulation of T cell autoimmunity . The effect of neutralizing antibodies on the sustainable efficacy of biologic therapies : what 's in it for African and Middle Eastern rheumatologists . Over the last decade , biologic therapeutic proteins have advanced the treatment of diseases such as rheumatoid arthritis ( RA ) . Therapeutic antibodies such as infliximab , adalimumab , rituximab , tocilizumab , DB06674 , certolizumab pegol , the receptor construct etanercept , and abatacept , an anticluster of differentiation (CD)80/anti- P42081 fusion protein , are used as treatment for RA and ankylosing spondylitis ( AS ) . DB00065 , adalimumab , DB06674 , certolizumab pegol , and etanercept are inhibitors of tumor necrosis factor ( P01375 ) , a key regulator of inflammation . Left untreated , progression of rheumatic diseases due to inflammation can lead to irreversible joint damage and serious disability . One limitation for the use of therapeutic antibodies is immunogenicity , the induction of antibodies by the adaptive immune system in response to foreign substances . The development of antidrug antibodies ( ADAs ) has a varying impact on the clinical efficacy of biologic agents for the treatment of RA and AS , depending on whether the ADAs are neutralizing or non-neutralizing . Studies have indicated that neutralizing ADAs are associated with a reduced efficacy , decreased drug survival , increased instances of dose escalation , and adverse events . Comparison studies of anti- P01375 biologics have demonstrated that each drug has a different sustained efficacy profile depending on immunogenicity . The purpose of this review is to provide rheumatologists with information regarding the effect of neutralizing antibodies on the sustainable efficacy of anti- P01375 biologic therapies . This information will be of value to practicing rheumatologists in Africa and the Middle East who should take into account the potential for changes in the efficacy and safety of biologic therapies and closely monitor patients under their care . DB00877 induces Q8NHJ6 (high) Q8N423 (high) dendritic cells promoting a new immunoregulatory pathway . Q8NHJ6 (high) Q8N423 (high) dendritic cells ( DCs ) may cause anergy in P01730 (+)CD45RO(+)CD25(+) T cells transforming them into regulatory T cells ( Tregs ) . Here , we tested whether chronic exposure to rapamycin may modulate this immunoregulatory pathway in renal transplant recipients . Forty renal transplant patients with biopsy-proven chronic allograft nephropathy and receiving calcineurin inhibitors were randomly assigned to either calcineurin inhibitor dose reduction or withdrawal with rapamycin introduction . At conversion and 2 years thereafter , we measured the rapamycin effects on circulating DCs ( BDCA1/ Q8WTT0 and Q8NHJ6 / Q8N423 expression ) , P01730 (+)/CD25(high)/Foxp3(+) Tregs , CD8(+)/ P10747 (-) T cells , and the Th1/Th2 balance in graft biopsies . In rapamycin-treated patients , peripheral Q8WTT0 (+) cells were significantly increased along with Q8NHJ6 / Q8N423 (+) DCs . The number of circulating P01730 (+)/CD25(high)/Foxp3(+)/ P16410 (+) Tregs , CD8(+) P10747 (-) T cells , and P17693 serum levels were higher in the rapamycin-treated group . The number of Q8NHJ6 / Q8N423 (+) Q8WTT0 (+) DC was directly and significantly correlated with circulating Tregs and CD8(+) P10747 (-) T cells . Q8NHJ6 / Q8N423 expression was increased in kidney biopsies at the end of the study period along with a significant bias toward a Th2 response within the graft only in the rapamycin-treated patients . Thus , rapamycin induces the upregulation of Q8NHJ6 and Q8N423 on the DC surface , and this effect is associated with an increase in the number of Tregs and expansion of the CD8(+) P10747 (-) T cell population . This suggests that P42345 inhibition may promote a novel immunoregulatory pathway . Apoptosis and the FLIP and NF-kappa B proteins as pharmacodynamic criteria for biosimilar P01375 antagonists . Various criteria are necessary to assess the efficacy and safety of biological medications in order to grant companies the right to register these medications with the appropriate bodies that regulate their sale . The imminent expiration of the patents on reference biological products which block the cytokine P01375 -α ( tumor necrosis factor-α ) raises the possibility of bringing so-called biosimilars to the market ( similar to the biologicals of reference products ) . This occurrence is inevitable , but criteria to adequately evaluate these medications are now needed . Even among controversy , there is a demand from publications correlating the pro-apoptotic mechanism of the original P01375 -α antagonists ( etanercept , infliximab , adalimumab , DB06674 , and certolizumab pegol ) in the treatment of rheumatoid arthritis and other diseases . In this article , the authors discuss the possibility of utilizing the pro-apoptotic effect correlated with the regulation of the anti-apoptotic proteins FLIP and NF-κB as new criteria for analyzing the pharmacodynamics of possible biosimilar P01375 -α antagonists which should be submitted to regulatory agencies for evaluation . Molecular and biologic characterization of a newly established Philadelphia-positive acute lymphoblastic leukemia cell line ( Z-33 ) with an autocrine response to GM- P04141 . We have recently established a new Philadelphia chromosome ( Ph1 ) -positive acute lymphoblastic leukemia ( ALL ) cell line , designated Z-33 . This line has Q401N2 morphology , ultrastructural characteristics of lymphoblasts and typical B lineage surface markers identical to those observed in the Ph1-positive ALL patient from whom the line was derived . In addition , a rearranged immunoglobulin heavy-chain gene ( JH ) band was found in Z-33 cells by Southern blot analysis , confirming B cell clonality . Cytogenetic analysis of the cell line revealed t(9;22)(q34;q11.2) . Polymerase chain reaction ( PCR ) -amplified cDNA from Z-33 cells demonstrated an e1-az P11274 - P00519 junction , and the p190BCR- P00519 protein was detected in them by the immune complex kinase assay . Z-33 cells produce interleukin ( IL ) -1 beta , P05231 , granulocyte colony-stimulating factor ( DB00099 ) , granulocyte-macrophage P04141 ( GM- P04141 ) , tumor necrosis factor ( P01375 ) -alpha , and transforming growth factor ( TGF ) -beta , Neither P01584 , DB00099 , P01375 , nor their corresponding antibodies affected the cell line 's growth . In contrast , anti-GM- P04141 neutralizing antibodies suppressed Z-33 colony formation , and GM- P04141 stimulated it in a dose-dependent fashion . In addition , receptor studies with biotinylated GM- P04141 demonstrated specific binding to Z-33 cells , indicating that the cells express GM- P04141 receptors . Taken together , our data suggest that the Ph1-positive Z-33 ALL cells produce GM- P04141 , express GM- P04141 receptors , and show an autocrine proliferative response to this cytokine . One target , different effects : a comparison of distinct therapeutic antibodies against the same targets . To date , more than 30 antibodies have been approved worldwide for therapeutic use . While the monoclonal antibody market is rapidly growing , the clinical use of therapeutic antibodies is mostly limited to treatment of cancers and immunological disorders . Moreover , antibodies against only five targets ( P01375 -α , P04626 , P11836 , P00533 , and P15692 ) account for more than 80 percent of the worldwide market of therapeutic antibodies . The shortage of novel , clinically proven targets has resulted in the development of many distinct therapeutic antibodies against a small number of proven targets , based on the premise that different antibody molecules against the same target antigen have distinct biological and clinical effects from one another . For example , four antibodies against P01375 -α have been approved by the FDA -- infliximab , adalimumab , DB06674 , and certolizumab pegol -- with many more in clinical and preclinical development . The situation is similar for P04626 , P11836 , P00533 , and P15692 , each having one or more approved antibodies and many more under development . This review discusses the different binding characteristics , mechanisms of action , and biological and clinical activities of multiple monoclonal antibodies against P01375 -α , HER-2 , P11836 , and P00533 and provides insights into the development of therapeutic antibodies . DB08904 in axial spondyloarthritis . The axial spondyloarthritis ( SpA ) classification criteria cover both patients with ankylosing spondylitis and non-radiographic axial SpA . After failure of NSAIDs P01375 -α-inhibitors ( P01375 -blockers ) can be given to patients with active axial SpA . Until recently , the P01375 -blockers infliximab , adalimumab , etanercept and DB06674 are labeled for the treatment of active ankylosing spondylitis while for active nr-axSpA only adalimumab has been approved in Europe . The P01375 -blocker certolizumab pegol has recently been evaluated in the RAPID-axSpA trial which is the first placebo-controlled randomized-controlled trial in the entire group of axial SpA . An elevated P02741 and/ or evidence of bone marrow edema on Q9BWK5 of the sacroiliac joints were required for inclusion in RAPID-axSpA , and patients could have been preexposed to P01375 -blockers . The interesting data of this important trial in the context of the emerging therapeutic field of non-radiographic axial SpA therapy is discussed in this review . Can we reduce the dosage of biologics in spondyloarthritis ? P01375 blockers have revolutionized the management of spondyloarthritis ( SpA ) . To date , four anti-TNFα agents ( etanercept , infliximab , adalimumab , DB06674 ) have been approved for the management of ankylosing spondylitis ( AS ) and psoriatic arthritis ( PsA ) . The first objective in the management of AS and PsA with P01375 inhibitors is to reduce disease activity to clinical remission or low disease activity . After remission has been achieved , this state should be maintained as long as possible . However , the financial burden associated with the cost of anti- P01375 agents as well as concerns about their long-term safety suggest reducing the dosage of the drug or discontinuing the therapy in the hopes of drug-free remission . The aim of this review is to examine what has , till now , been published on this topic in axial SpA , which includes AS and non-radiographic axial SpA ( nr-axSpA ) , peripheral SpA and PsA . Discontinuation of therapy in axial SpA is not possible in the majority of patients , while on the contrary , reducing the dosage often is . In some patients with peripheral SpA and PsA it is also possible to discontinue therapy and to achieve drug-free remission . Pharmacogenetics and future therapeutic scenarios : what affects the prediction of response to treatment with etanercept ? There are five tumor necrosis factor alpha ( P01375 -α ) inhibitors available for clinical use that have demonstrated efficacy as monotherapy or in combination with other anti-inflammatory or disease-modifying anti-rheumatic drugs ( DMARDs ) in the treatment of immune-mediated diseases . These include the anti- P01375 -α monoclonal antibodies infliximab , adalimumab , DB06674 , and certolizumab pegol , and the fusion protein , etanercept . The use of pharmacogenetic testing has the potential to increase drug efficiency by identifying genetic factors responsible for a lack of response to , or toxicities from , P01375 -α inhibitors , and could be used to individualize therapy . Several studies have reported associations between genetic polymorphisms and the response to etanercept , but most are small and insufficiently powered to detect effect , and markers tend to be more prognostic than predictive of therapeutic response . Limitations of pharmacogenetic studies include the use of single nucleotide polymorphisms ( SNPs ) , genes in linkage with other loci , interaction of environmental factors , and cohort heterogeneity , all of which can complicate the relationship between genetic polymorphisms and treatment response . Further studies are needed for pharmacogenetics to become a routine part of daily clinical therapeutic practice . P01375 inhibitors - state of knowledge . P01375 ( P01375 ) is considered a major proinflammatory cytokine , affecting various aspects of the immune reaction . All five P01375 inhibitors currently available on the market ( i.e. , etanercept , infliximab , adalimumab , certolizumab and DB06674 ) are top sellers , although indicated only in autoimmune diseases , including rheumatoid arthritis , Crohn 's disease and psoriasis . This article briefly discusses the background and place for P01375 inhibitors in modern therapy . The main safety aspects of P01375 inhibitor administration are described in particular , with special consideration of the available meta-analyses . Finally , perspectives on the next-generation P01375 inhibitors and their use in the clinic are given . DB00877 antagonizes P01375 induction of P19320 on endothelial cells by inhibiting mTORC2 . Recruitment of circulating leukocytes into inflamed tissues depends on adhesion molecules expressed by endothelial cells ( ECs ) . Here we report that rapamycin pretreatment reduced the ability of P01375 -treated ECs to capture T cells under conditions of venular flow . This functional change was caused by inhibition of P01375 -induced expression of vascular cell adhesion molecule-1 ( P19320 ) and could be mimicked by knockdown of mammalian target of rapamycin ( P42345 ) or rictor , but not raptor , implicating mTORC2 as the target of rapamycin for this effect . Mechanistically , mTORC2 acts through Akt to repress Raf1- Q02750 /2- P27361 /2 signaling , and inhibition of mTORC2 consequently results in hyperactivation of P27361 /2 . Increased P27361 /2 activity antagonizes P19320 expression by repressing P01375 induction of the transcription factor P10914 . Preventing activation of P27361 /2 reduced the ability of rapamycin to inhibit P01375 -induced P19320 expression . In vivo , rapamycin inhibited mTORC2 activity and potentiated activation of P27361 /2 . These changes correlated with reduced endothelial expression of P01375 -induced P19320 , which was restored via pharmacological inhibition of P27361 /2 . Functionally , rapamycin reduced infiltration of leukocytes into renal glomeruli , an effect which was partially reversed by inhibition of P27361 /2 . These data demonstrate a novel mechanism by which rapamycin modulates the ability of vascular endothelium to mediate inflammation and identifies endothelial mTORC2 as a potential therapeutic target . Pharmacology of P01375 blockade in rheumatoid arthritis and other chronic inflammatory diseases . P01375 -alpha ( P01375 ) has been unequivocally validated as a therapeutic target in a number of immune-mediated inflammatory disorders ( IMIDs ) . There is now increasing choice of biologic agents within the class all of which successfully neutralize sTNF . But approaches to P01375 inhibition differ and currently include mAbs ( infliximab , adalimumab , and DB06674 ) , either chimeric or human in sequence , a PEGylated Fab ' fragment ( certolizumab ) , and an IgG1- P20333 fusion protein ( etanercept ) . It is emerging that the pharmacological properties of these three anti- P01375 subtypes differ with respect to Fc function , binding of tmTNF and the possible consequences of this , as well as the ability to form complexes . The mode of administration of each agent , clearance and the local tissue concentrations achieved may also confer unique characteristics of relevance with respect to efficacy and safety . DB00278 -coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 has a high affinity for thrombin and its thrombin binding is reversible . P00734 derived from a Ba(2+) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days . Thalidomide suppresses Up-regulation of human immunodeficiency virus coreceptors P61073 and P51681 on P01730 + T cells in humans . Concurrent infection in patients with human immunodeficiency virus ( HIV ) infection increases the expression of HIV coreceptors P61073 and P51681 . Thalidomide has beneficial effects in a number of HIV-associated diseases . The effect of thalidomide on P61073 and P51681 expression on P01730 + T cells was determined . Thalidomide produced a dose-dependent inhibition of lipopolysaccharide ( LPS ) -induced up-regulation of P61073 and P51681 in vitro . Antibody to tumor necrosis factor-alpha ( P01375 ) also attenuated the LPS-induced HIV coreceptor up-regulation , which was not further reduced by thalidomide . Thalidomide ( 400 mg ) was orally administered to 6 men , and their blood was stimulated ex vivo with LPS , staphylococcal or mycobacterial antigens , or antibody to CD3 or P10747 cells . All stimuli induced up-regulation of HIV coreceptors , which was reduced after ingestion of thalidomide . Thalidomide may be beneficial in the treatment of intercurrent infections during HIV infection by reducing the up-regulation of P61073 and P51681 expression on P01730 + T cells induced by bacterial and mycobacterial antigens , by a mechanism that involves inhibition of P01375 . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Effect of interferon-gamma and P01375 on P15941 mucin expression in ovarian carcinoma cell lines . In view of the potential uses of cell surface tumour associated antigens in novel anticancer treatment , a study was designed to investigate whether the biological response modifiers interferon-gamma ( P01579 ) and tumour necrosis factor-alpha ( P01375 ) could effect the expression of an epitope on the tumour associated P15941 epithelial mucin . Four ovarian carcinoma cell lines showing high ( OAW42 and GG ) and low ( JAM and PE01 ) basal expression of P15941 were treated with 10-1000 U/mL of P01579 or P01375 for one or five days . Changes in P15941 expression in cells exposed to P01579 or P01375 were monitored using an ELISA technique with the monoclonal antibody O43633 which reacts with a core protein epitope on the P15941 mucin , and then corrected for the number of viable cells present . P01375 had little effect on P15941 expression , but one or five days exposure to P01579 significantly increased P15941 expression ( p < 0.01 ) in all cell lines including the two cell lines that initially showed little or no expression . Spondyloarthritides . The most important clinical features of the spondyloarthritides ( SpA ) are not only inflammatory back pain ( Q9H4E7 ) but also peripheral ( enthesitis ) and extra-articular symptoms . For clinical purposes , two forms related to the predominant clinical manifestation - axial and peripheral SpA - and five subgroups- ankylosing spondylitis ( AS ) , SpA associated with psoriasis and inflammatory bowel disease ( Q9UKU7 ) , reactive arthritis and undifferentiated SpA - are differentiated . Axial SpA including AS is the most frequent subtype of SpA , followed by psoriatic arthritis and undifferentiated SpA , while reactive arthritis and Q9UKU7 -related SpA are less frequent . The prevalence of SpA has been shown to be similar to rheumatoid arthritis . The outcome of the disease is influenced by the degree of disease activity over time , which is mainly related not only to inflammation but also on the structural damage ( new bone formation ) that occurs over time . Treatment options for patients with SpA have been limited for decades . Non-steroidal anti-inflammatory agents are currently considered first choice , since they have shown good amelioration of symptoms in SpA patients especially when suffering by the typical symptom of Q9H4E7 . Furthermore , there is a clear role for regular physiotherapy in AS to prevent loss of spinal mobility . For patients who have insufficiently responded to conventional therapies , four anti-tumour necrosis factor ( P01375 ) agents are available and are approved for the treatment of patients with active AS : infliximab , etanercept , adalimumab and DB06674 . As far as it stands now , P01375 blockers seem to have no influence on new bone formation in AS . New treatments for inflammatory rheumatic disease . As our understanding of the pathogenesis of autoimmune diseases is growing , new therapies are being developed to target disease-specific pathways . Since the introduction of etanercept in 1998 , several biotechnological agents have been developed , most of them indicated in the treatment of rheumatoid arthritis , but also psoriatic arthritis . Most currently available molecules target P01375 -alfa with different strategies ( i.e. , etanercept , infliximab , adalimumab , DB06674 , and certolizumab pegol ) , P05231 ( tocilizumab ) , P16410 ( abatacept ) , and B cells ( rituximab , belimumab ) as they are key mediators in the cascade of inflammation . Further , small molecules have been recently developed to target intracellular signaling , such as Janus Kinases for tofacitinib , the first FDA-approved small molecule for rheumatoid arthritis . Most novel treatments are being developed for arthritis with specific differences between rheumatoid and psoriatic arthritis , as well as for systemic lupus erythematosus , following the approval of belimumab . Finally , biologic therapies are effective also in gout , mainly targeting interleukin-1 to block the inflammasome . This review article describes the new and upcoming treatment options for rheumatoid arthritis , psoriatic arthritis , systemic lupus erythematosus , and gout to dissect what we should be aware of when discussing these new and promising molecules . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . Biologics in the management of ulcerative colitis - comparative safety and efficacy of P01375 -α antagonists . Ulcerative colitis can cause debilitating symptoms and complications such as colonic strictures , colonic dysplasia , colorectal cancer , and toxic megacolon or perforation . Goals of treatment in ulcerative colitis include resolution of gastrointestinal symptoms , healing of colonic mucosa , and prevention of disease complications . Our treatment armamentarium has expanded dramatically over the past 10 years , and we now have multiple biologic agents approved for the treatment of moderate-severe disease , in addition to conventional therapies such as 5-aminosalicylates , thiopurines , and corticosteroids . In this review , we will provide a detailed discussion of the three tumor necrosis factor-alpha ( P01375 -α ) inhibitors currently approved for treatment of ulcerative colitis : infliximab , adalimumab , and DB06674 . All three agents are effective for inducing and maintaining clinical response and remission in patients with ulcerative colitis , and they have comparable safety profiles . There are no head-to-head trials comparing their efficacy , and the choice of agent is most often based on insurance coverage , route of administration , and patient preference . Combination therapy with an immunomodulator is proven to be more effective than anti- P01375 monotherapy , and patients who lose response to an anti- P01375 agent should undergo dose intensification in order to regain clinical response . Despite therapeutic optimization , a significant percentage of patients will not achieve clinical remission with anti- P01375 agents , and so newer therapies are on the horizon .

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Suppressed Ca2+/ P62158 /CaMKII-dependent K( DB00171 ) channel activity in primary afferent neurons mediates hyperalgesia after axotomy . Painful axotomy decreases K( DB00171 ) channel current ( IK( DB00171 ) ) in primary afferent neurons . Because cytosolic Ca(2+) signaling is depressed in injured dorsal root ganglia ( Q86YR7 ) neurons , we investigated whether Ca(2+)-calmodulin ( P62158 ) -Ca(2+)/ P62158 -dependent kinase II ( CaMKII ) regulates IK( DB00171 ) in large Q86YR7 neurons . Immunohistochemistry identified the presence of K( DB00171 ) channel subunits Q09428 , SUR2 , and Kir6.2 but not Kir6.1 , and pCaMKII in neurofilament 200-positive Q86YR7 somata . Single-channel recordings from cell-attached patches revealed that basal and evoked IK( DB00171 ) by ionomycin , a Ca(2+) ionophore , is activated by CaMKII . In axotomized neurons from rats made hyperalgesic by spinal nerve ligation ( Q16658 ) , basal K( DB00171 ) channel activity was decreased , and sensitivity to ionomycin was abolished . Basal and Ca(2+)-evoked K( DB00171 ) channel activity correlated inversely with the degree of hyperalgesia induced by Q16658 in the rats from which the neurons were isolated . Inhibition of IK( DB00171 ) by glybenclamide , a selective K( DB00171 ) channel inhibitor , depolarized resting membrane potential ( O94763 ) recorded in perforated whole-cell patches and enhanced neurotransmitter release measured by amperometry . The selective K( DB00171 ) channel opener diazoxide hyperpolarized the O94763 and attenuated neurotransmitter release . Axotomized neurons from rats made hyperalgesic by Q16658 lost sensitivity to the myristoylated form of autocamtide-2-related inhibitory peptide ( AIPm ) , a pseudosubstrate blocker of CaMKII , whereas axotomized neurons from Q16658 animals that failed to develop hyperalgesia showed normal IK( DB00171 ) inhibition by AIPm . AIPm also depolarized O94763 in control neurons via K( DB00171 ) channel inhibition . Unitary current conductance and sensitivity of K( DB00171 ) channels to cytosolic DB00171 and ligands were preserved even after painful nerve injury , thus providing opportunities for selective therapeutic targeting against neuropathic pain . Skinned coronary smooth muscle : calmodulin , calcium antagonists , and DB02527 influence contractility . The effects of Ca2+ , calmodulin , DB02527 , the catalytic subunit of DB02527 -dependent protein kinase ( CSU ) and some Ca2+ antagonists were studied in chemically ( Triton X-100 ) skinned coronary smooth muscle . P62158 increased the Ca2+ responsiveness of the muscle fiber as indicated by the reduction in the threshold as well as the half-maximal activating Ca2+ concentration . DB00831 , a calmodulin antagonist , inhibited Ca2+-calmodulin-induced contraction . Both DB02527 and CSU were effective inhibitors of contraction induced at an intermediate Ca2+ concentration . DB08980 , a Ca2+-antagonist , at 2 x 10(-4) M produced a significant inhibitory effect , which was reduced by increasing the Ca2+ concentration . From other Ca2+ antagonists tested , W-7 , but not D600 and verapamil , produced some inhibitory effect . The data indicate that the response of skinned coronary smooth muscle to Ca2+ , calmodulin and DB02527 are similar to those obtained with other skinned smooth muscles . Furthermore , skinned fiber preparation can serve as a useful tool to investigate possible direct effects of drugs on the activating and regulatory systems in smooth muscle . P15056 inhibitors suppress apoptosis through off-target inhibition of JNK signaling . DB08881 and dabrafenib selectively inhibit the P15056 ( P15056 ) kinase , resulting in high response rates and increased survival in melanoma . Approximately 22 % of individuals treated with vemurafenib develop cutaneous squamous cell carcinoma ( cSCC ) during therapy . The prevailing explanation for this is drug-induced paradoxical P29323 activation , resulting in hyperproliferation . Here we show an unexpected and novel effect of vemurafenib/PLX4720 in suppressing apoptosis through the inhibition of multiple off-target kinases upstream of c-Jun N-terminal kinase ( JNK ) , principally Q9NYL2 . JNK signaling is suppressed in multiple contexts , including in cSCC of vemurafenib-treated patients , as well as in mice . Expression of a mutant Q9NYL2 that can not be inhibited reverses the suppression of JNK activation and apoptosis . Our results implicate suppression of JNK-dependent apoptosis as a significant , independent mechanism that cooperates with paradoxical P29323 activation to induce cSCC , suggesting broad implications for understanding toxicities associated with P15056 inhibitors and for their use in combination therapies . DOI : http://dx.doi.org/10.7554/eLife.00969.001 . Blood flow alterations in TNBS-induced colitis : role of endothelin receptors . OBJECTIVES : The aim of the present study was to investigate the time dependent changes in hemodynamic parameters and to assess the role of endothelin ( ET ) receptors in trinitrobenzene sulfonic acid ( TNBS ) induced colitis . MATERIALS : Inferior mesenteric artery ( IMA ) hemodynamics , myeloperoxidase activity ( P05164 ) and damage scores were measured immediately or 1 , 3 , 5 and 14 days after colitis . TREATMENTS : Another group of rats received a nonselective ET receptor antagonist DB00559 ( 30 mg/kg/day ) , P25101 receptor antagonist BQ485 ( 60 microg/rat/day ) or P24530 receptor antagonist BQ788 ( 60 microg/rat/day ) prior to and on the 1st , 2nd and 3rd days after TNBS administration . RESULTS : IMA flow significantly increased at 90 min followed by a substantial decrease through days 1-5 . Tissue P05164 activity and macroscopic damage score increased on 1st day after the induction of colitis and remained elevated 3 , 5 and 14 days following colitis . Treatment with DB00559 or P25101 receptor antagonist largely prevented the colitis-induced reduction in blood flow and tissue injury whereas P24530 receptor antagonist did not attenuate tissue injury or reductions in blood flow . CONCLUSIONS : Our results demonstrate that time-dependent abnormalities occur in IMA hemodynamics following TNBS administration . Our findings also indicate that P25101 receptors but not P24530 receptors play an important role in the colonic inflammation following TNBS administration . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . Endothelins stimulate sodium uptake into rat brain capillary endothelial cells through endothelin A-like receptors . The effect of endothelins ( ETs ) on sodium/hydrogen ( Na+/H+ ) antiport system was examined in cultured rat brain capillary endothelium ( RBEC ) . ET-1 , P20800 , and P14138 stimulated Na+ uptake into RBEC with similar half-maximal stimulation ( EC50 ) values ( 0.7 , 0.6 , and 1.1 nM , respectively ) . This reaction was inhibited by the Na+/H+ antiport inhibitor , N-(ethyl-N-isopropyl)-amiloride ( EIPA ) . The selective endothelin A ( P25101 ) receptor-antagonist ( cyclo-D- DB00150 -D- DB00128 -Pro-D- DB00161 - DB00149 ( BQ123 ) ) , but not endothelin B ( ETB ) receptor-antagonists ( ( Cys11 , Cys15 ) -ET-1 ( IRL1038 ) or N-cis-2,6-dimethylpiperidinocarbonyl-L-gamma MeLeu-D- DB00150 (COOMe)-D-Nle-ONa ( BQ788 ) ) , inhibited both ET-1- and P14138 -stimulated Na+ uptake , indicating P25101 -receptor mediation . The protein kinase C ( PKC ) activator ( phorbol 12-myristate 13-acetate ( PMA ) ) failed to stimulate Na+ uptake . The calcium-calmodulin ( P62158 ) inhibitor ( W7 ) reduced ET-1-stimulated Na+ uptake by 50 % , whereas the PKC inhibitor ( staurosporine ) had no effect , indicating that ET-1 stimulation of the Na+/H+ antiport system is linked to a P62158 -dependent and PKC-independent pathway . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . Resistance of P04626 /neu-overexpressing tumor targets to lymphokine-activated-killer-cell-mediated lysis : evidence for deficiency of binding and post-binding events . P04626 /neu-overexpressing tumor cell lines are relatively resistant to lymphokine-activated killer ( Q96QP1 ) cell cytotoxicity when compared to P04626 /neu-nonexpressing lines . P04626 /neu+ targets were also resistant to binding by Q96QP1 large granular lymphocytes ( LGL ) as shown by visualization at the single-cell level , a target monolayer binding assay and in " cold " target inhibition experiments . P04626 /neu+ Q96QP1 -resistant ovarian cell lines demonstrated an absence of P05362 expression while expression of LFA-3 , N- P62158 , laminin and beta 1 integrins was comparable to that of P04626 /neu- targets . In contrast , the P04626 /neu+ breast cell line , SKBR-3 , which was also resistant to lysis and binding by Q96QP1 LGL , demonstrated normal expression of P05362 . Anti- P05362 antibodies blocked binding and lysis of P04626 /neu- carcinoma targets by Q96QP1 cells , further supporting the notion that lack of P05362 expression on P04626 /neu+ cells contributes to their resistance . The modest binding and lysis of P04626 /neu+ targets by Q96QP1 cells was significantly inhibited by anti-LFA-1 antibodies , suggesting the existence of another counter-receptor for LFA-1 on P04626 /neu+ targets . The following also supported deficiencies in post-binding events when P04626 /neu+ cells resisted the lytic activity of Q96QP1 cells : ( a ) when the relative resistance to effector cell binding was overcome by exogenous lectin . P04626 /neu+ cell lines were still resistant to Q96QP1 cytolysis , and ( b ) P04626 /neu+ targets were resistant to perforin-containing granule extracts obtained from the CTLL-R8 cytotoxic lymphocyte cell line . These results indicate that deficiency in effector binding as well as post-binding events contributes to the resistance of P04626 /neu-overexpressing tumor targets to Q96QP1 -cell-mediated lysis . Bioassay-guided isolation of an alkaloid with antiangiogenic and antitumor activities from the extract of Fissistigma cavaleriei root . Fissistigma cavaleriei ( Levl ) Rehd ( Annonaceae ) is used as a folklore medicine for treatment of inflammation , arthritis , and tuberculosis by Miao people in China . In the present study , the antiangiogenic activity of F. cavaleriei was investigated . The chorioallantoic membrane of the fertilized hen 's egg ( P62158 assay ) was used to determine antiangiogenic activity of the plant extract . Compound ( 1 ) , a compound with antiangiogenic activity , was isolated by bioassay-guided fractionation from F. cavaleriei for the first time . The structure of compound ( 1 ) was elucidated on the basis of spectroscopic methods . Colorimetric P36551 ( ovine ) inhibitor screening assay was used to determine its inhibitory effect on P23219 and P35354 . MTT and Sulforhodamine B assays were used to investigate its cytotoxic effects on tumor cell lines . As a result , compound ( 1 ) showed a selectively inhibiting effect on P35354 and could inhibit the growth of tumor cells in vitro . The antitumor activity of compound ( 1 ) was further confirmed by the observation that compound ( 1 ) administration significantly inhibited the growth of S-180 cells in mice . Moreover , compound ( 1 ) was able to enhance the antitumor activity of doxorubicin in the mice bearing with S-180 cells while combined with doxorubicin . In conclusion , compound ( 1 ) is a multi-target molecule and further experimental investigations are needed to determine whether it can be used as a lead molecule for tumor treatment . DB06589 inhibits the activation of P09619 β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 -transfected MDA-MB-231 human breast carcinoma cells ( 231-BR- P04626 ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751- P09619 β ) was identified around perivascular brain micrometastases . p751- P09619 β(+) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231-BR- P04626 large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 treatment resulted in 70 % ( P = 0.023 ) decrease of the p751- P09619 β(+) astrocyte population , at the lowest dose of 30 mg/kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients . [ P62158 -dependent regulation of Ca,Mg-ATPase activity in plasma membranes of the swine myometrium ] . Highly purified plasma membrane ( PM ) preparations of pig myometrium were found to contain 0.91 +/- 0.22 microgram calmodulin per mg of PM protein . Treatment of membranes with 1 mM EGTA in the presence of 0.2 M NaCl causes the diminution of the calmodulin content down to 3 % of the original level . The activity of Ca , Mg-ATPase is thereby decreased by 40 % . Exogenous calmodulin restores the enzyme activity up to 1.94 +/- +/- 0.30 mumol Pi/mg protein/hour . The maximal activation of Ca , Mg-ATPase is observed with 10(-7) M calmodulin . P62158 increases the total ATPase activity of myometrium PM without affecting the Mg-ATPase activity . DB00831 ( 20 microM ) diminishes the activating effect of exogenous calmodulin on Ca , Mg-ATPase . P62158 stimulates Ca , Mg-ATPase at low concentrations of Ca2+ ( 10(-8)-10(-6) M ) by decreasing Km for Ca2+ from 0.4.10(-6) M to 2.10(-8) M as well as by increasing Vmax -- from 0,8 to 1.42 mumol Pl/mg protein/hour . It is supposed that the activating effect of calmodulin on Ca , Mg-ATPase is based on electrostatic interactions of Ca2+-free calmodulin with the enzyme . Mutation of the calmodulin binding motif IQ of the L-type Ca(v)1.2 Ca2+ channel to EQ induces dilated cardiomyopathy and death . Cardiac excitation-contraction coupling ( EC coupling ) links the electrical excitation of the cell membrane to the mechanical contractile machinery of the heart . DB01373 channels are major players of EC coupling and are regulated by voltage and Ca(2+)/calmodulin ( P62158 ) . P62158 binds to the IQ motif located in the C terminus of the Ca(v)1.2 channel and induces Ca(2+)-dependent inactivation ( CDI ) and facilitation ( P05231 ) . Mutation of DB00167 to DB00142 ( Ile1624Glu ) in the IQ motif abolished regulation of the channel by CDI and P05231 . Here , we addressed the physiological consequences of such a mutation in the heart . Murine hearts expressing the Ca(v)1.2(I1624E) mutation were generated in adult heterozygous mice through inactivation of the floxed WT Ca(v)1.2( Q401N2 ) allele by tamoxifen-induced cardiac-specific activation of the MerCreMer Cre recombinase . Within 10 days after the first tamoxifen injection these mice developed dilated cardiomyopathy ( DCM ) accompanied by apoptosis of cardiac myocytes ( CM ) and fibrosis . In Ca(v)1.2(I1624E) hearts , the activity of phospho- P62158 kinase II and phospho-MAPK was increased . CMs expressed reduced levels of Ca(v)1.2(I1624E) channel protein and I(Ca) . The Ca(v)1.2(I1624E) channel showed " CDI " kinetics . Despite a lower sarcoplasmic reticulum Ca(2+) content , cellular contractility and global Ca(2+) transients remained unchanged because the EC coupling gain was up-regulated by an increased neuroendocrine activity . Treatment of mice with metoprolol and captopril reduced DCM in Ca(v)1.2(I1624E) hearts at day 10 . We conclude that mutation of the IQ motif to IE leads to dilated cardiomyopathy and death . Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 ) , the regulatory subunit of the NCCa- DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) /reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson׳s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 mRNA and protein were maximally upregulated 8-12h after a 2-hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 -regulated NCCa- DB00171 channel may be associated with MCAO/reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain . Gambogic acid inhibits angiogenesis through suppressing vascular endothelial growth factor-induced tyrosine phosphorylation of P35968 /Flk-1 . Previous studies revealed that gambogic acid ( GA ) , the major active ingredient of gamboge , a brownish to orange resin exuded from Garcinia hanburryi tree in Southeast Asia , possessed significant anticancer activity both in vitro and in vivo . In this study , we explored the high antiangiogenic activities of GA for the first time . GA inhibits the P15692 -stimulated proliferation , migration and tube formation of human umbilical vein endothelial cells ( HUVECs ) as well as microvessel sprouting from rat aortic rings in vitro . Moreover , GA inhibits vessel growth in matrigel plugs and P62158 in vivo and transplanted tumor in mice . The results also indicated that GA decreases P15692 production of cultured tumor cells and inhibits P15692 -induced tyrosine phosphorylation of P35968 /Flk-1 . This inhibition of receptor phosphorylation is correlated with a significant decrease in P15692 -triggered phosphorylated forms of P29323 , AKT and p38 . Taken together , these findings strongly suggest that GA might be a structurally novel angiogenesis inhibitor . Products of the unc-52 gene in Caenorhabditis elegans are homologous to the core protein of the mammalian basement membrane heparan sulfate proteoglycan . Mutations in the unc-52 gene of Caenorhabditis elegans affect attachment of the myofilament lattice to the muscle cell membrane . Here , we demonstrate that the unc-52 gene encodes a nematode homolog of perlecan , the mammalian basement membrane heparan sulfate proteoglycan . The longest potential open reading frame of this gene encodes a 2482-amino-acid protein with a signal peptide and four domains . The first domain is unique to the unc-52 polypeptide , whereas the three remaining domains contain sequences found in the P01130 ( domain II ) laminin ( domain III ) and N- P62158 ( domain IV ) . We have identified three alternatively spliced transcripts that encode different carboxy-terminal sequences . The two larger transcripts encode proteins containing all or part of domain IV , whereas the smaller transcript encodes a shortened polypeptide that completely lacks domain IV . We have determined that the disorganized muscle phenotype observed in unc-52(st196) animals is caused by the insertion of a Tc1 transposon into domain IV . Two monoclonal antibodies that recognize an extracellular component of all contractile tissues in C. elegans fail to stain embryos homozygous for a lethal unc-52 allele . We have mapped the epitopes recognized by both monoclonal antibodies to a region of domain IV in the unc-52-encoded protein sequence . Therapy with a synthetic retinoid -- ( Ro 10-1670 ) etretin -- increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 ) -and retinoic acid ( CRABP ) -binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 levels were not altered during therapy . The results show that P09455 and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Traumatic brain injury-induced acute gene expression changes in rat cerebral cortex identified by GeneChip analysis . Proper CNS function depends on concerted expression of thousands of genes in a controlled and timely manner . Traumatic brain injury ( TBI ) in mammals results in neuronal death and neurological dysfunction , which might be mediated by altered expression of several genes . By employing a CNS-specific GeneChip and real-time polymerase chain reaction ( PCR ) , the present study analyzed the gene expression changes in adult rat cerebral cortex in the first 24 hr after a controlled cortical impact injury . Many functional families of genes not previously implicated in TBI-induced brain damage are altered in the injured cortex . These include up-regulated transcription factors ( O14543 , O60674 , P35610 -3 , Q03060 , P10914 , SMN , silencer factor-B , ANIA-3 , ANIA-4 , and DB09106 -1 ) and signal transduction pathways ( cpg21 , Narp , and P09455 ) and down-regulated transmitter release mechanisms ( CITRON , synaptojanin II , ras-related rab3 , neurexin-1beta , and SNAP25A and -B ) , kinases ( IP-3-kinase , Pak1 , Ca(2+)/ P62158 -dependent protein kinases ) , and ion channels ( K(+) channels TWIK , RK5 , X62839 , and Na(+) channel I ) . In addition , several genes previously shown to play a role in TBI pathophysiology , including proinflammatory genes , proapoptotic genes , heat shock proteins , immediate early genes , neuropeptides , and glutamate receptor subunits , were also observed to be altered in the injured cortex . Real-time PCR analysis confirmed the GeneChip data for many of these transcripts . The novel physiologically relevant gene expression changes observed here might explain some of the molecular mechanisms of TBI-induced neuronal damage . [ Functional characteristics of calcium-sensitive adenylyl cyclase of ciliate Tetrahymena pyriformis ] . DB01373 -sensitive forms of adenylyl cyclase ( AC ) were revealed in most vertebrates and invertebrates and also in some unicellular organisms , in particular ciliates . We have shown for the first time that calcium cations influence the AC activity of ciliate Tetrahymena pyriformis . These cations at the concentrations of 0.2-20 microM stimulated the enzyme activity , and maximum of catalytic effect was observed at 2 microM Ca2+ . DB01373 cations at a concentrations of 100 microM or higher inhibited the AC activity . P62158 antagonists W-5 and W-7 at the concentrations of 20-100 microM inhibited the catalytic effect induced by 5 microM Ca2+ and blocked the effect at higher concentrations of Ca2+ . DB00477 , another calmodulin antagonist , reduced Ca2+-stimulated AC activity only at the concentrations of 200-1000 microM . AC stimulating effects of serotonin , P01133 and DB02527 increased in the presence of 5 microM Ca2+ . AC stimulating effects of P01133 , DB02527 and insulin decreased in the presence of 100 microM Ca2+ , and AC stimulating effect of DB02527 decreased also in the presence of calmodulin antagonists ( 1 mM ) . At the same time , stimulating effect of D-glucose in the presence of Ca2+ and calmodulin antagonists did not change essentially . The data obtained speak in favor of the presence of calcium-sensitive forms of AC in ciliate T. pyriformis which mediate enzyme stimulation by P01133 , DB02527 , insulin , and serotonin .

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interacts_with DB01235?

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Comparison of rating scales used to evaluate DB01235 -induced dyskinesia in the 6-OHDA lesioned rat . Abnormal involuntary movement ( AIM ) rating scales are frequently used to study the mechanisms underlying DB01235 -induced dyskinesia ( LID ) in 6-OHDA lesioned rodents and the propensity of novel treatments for Parkinson 's disease to induce or alleviate similar abnormal behaviours . Despite the existence of at least one well validated method , other AIM scales are also in use . Moreover , there have been developments and variations in the original scales and their methods of use , without re-validation . In this study , 6-OHDA medial forebrain bundle lesioned Sprague-Dawley rats were treated with chronic DB01235 6 mg/kg/day for 5 weeks followed by 12 mg/kg/day for another 5 weeks . Rats were assessed weekly by simultaneous ratings on four published AIM and stereotypy scales with concurrent recording of rotation , over 3 hours following DB01235 injection . Three contemporary AIM scales have then been validated pharmacologically using agents that are known to reduce LID clinically and in primates ( amantadine ) or to interfere with the activity of DB01235 ( the D(1) and P14416 antagonists , P35240 -23390 and raclopride ) respectively . We also demonstrate that AIM , stereotypic and rotational behaviour are distinct motor dysfunctions induced by chronic and acute treatment of DB01235 , and should be assessed separately . The undertaking of assessments at multiple time points is essential especially when testing the efficacy of new potential anti-dyskinetic treatments . Importantly critical to all AIM and rotation testing is the internal validation of both the scale being used and the environment being used . Potentiator ivacaftor abrogates pharmacological correction of ΔF508 P13569 in cystic fibrosis . Cystic fibrosis ( CF ) is caused by mutations in the CF transmembrane conductance regulator ( P13569 ) . Newly developed " correctors " such as DB09280 ( VX-809 ) that improve P13569 maturation and trafficking and " potentiators " such as ivacaftor ( VX-770 ) that enhance channel activity may provide important advances in CF therapy . Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation ( G551D ) that affects only channel activity , a single compound is not sufficient to treat patients with the more common P13569 mutation , ΔF508 . Thus , patients with ΔF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit . However , whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro , the impact of chronic therapy has not been established . In studies of human primary airway epithelial cells , we found that both acute and chronic treatment with VX-770 improved P13569 function in cells with the G551D mutation , consistent with clinical studies . In contrast , chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated P13569 correction in ΔF508 homozygous cultures . This result reflected the destabilization of corrected ΔF508 P13569 by VX-770 , markedly increasing its turnover rate . Chronic VX-770 treatment also reduced mature wild-type P13569 levels and function . These findings demonstrate that chronic treatment with P13569 potentiators and correctors may have unexpected effects that can not be predicted from short-term studies . Combining these drugs to maximize rescue of ΔF508 P13569 may require changes in dosing and/or development of new potentiator compounds that do not interfere with P13569 stability . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . Detection of anti-isoniazid and anti-cytochrome P450 antibodies in patients with isoniazid-induced liver failure . Isoniazid ( DB00951 ) -induced hepatotoxicity remains one of the most common causes of drug-induced idiosyncratic liver injury and liver failure . This form of liver injury is not believed to be immune-mediated because it is not usually associated with fever or rash , does not recur more rapidly on rechallenge , and previous studies have failed to identify anti- DB00951 antibodies ( Abs ) . In this study , we found Abs present in sera of 15 of 19 cases of DB00951 -induced liver failure . Anti- DB00951 Abs were present in 8 sera ; 11 had anti-cytochrome P450 (CYP)2E1 Abs , 14 had Abs against P05181 modified by DB00951 , 14 had anti- P08684 antibodies , and 10 had anti- P11712 Abs . DB00951 was found to form covalent adducts with P05181 , P08684 , and P11712 . None of these Abs were detected in sera from DB00951 -treated controls without significant liver injury . The presence of a range of antidrug and autoAbs has been observed in other drug-induced liver injury that is presumed to be immune mediated . CONCLUSION : These data provide strong evidence that DB00951 induces an immune response that causes DB00951 -induced liver injury . P28702 polymorphisms do not explain functional differences in vitamins D and A response in Antineutrophil cytoplasmic antibody associated vasculitis patients . It has been suggested that the retinoid X receptor beta ( P28702 ) gene is a risk factor for Wegener 's granulomatosis . We addressed if there is a functional difference in the response to retinoic acid ( RA ) and vitamin D in Antineutrophil cytoplasmic antibody ( ANCA ) associated systemic vasculitis ( AASV ) patients and if this was associated with P28702 genotypes . TNFalpha and P22301 production were measured in whole blood assay from AASV patients ( n = 51 ) and healthy controls ( HC , n = 67 ) . One micromolar of 1,25-(OH)(2) D3 , 9-cis RA ( 9c-RA ) or all-trans RA ( DB00755 ) was added to the assay . Genotyping was performed for exons 7 and 2 of the P28702 gene and for a microsatellite in vicinity of the P28702 gene . Lipopolysaccharide ( LPS ) mediated TNFalpha production and P22301 were significantly lower in patients . Addition of 1,25-(OH)(2) D3 , DB00755 or 9c-RA , blunted TNFalpha production , more pronounced in patients . Although all three compounds inhibited P22301 production significantly in HC , only 1,25-(OH)(2) D3 was found to be effective in patients . Allele distribution of the P28702 microsatellite differed significantly between patients and HC . This was not found for the SNP in exons 2 and 7 . Genotype of the latter correlated with the ability of 1,25-(OH)(2) D3 and DB00755 to inhibit P22301 production . We provide immunological evidence for a functional difference in vitamins D and A responsiveness in AASV patients . Since the inhibition of TNFalpha was more effective in patients , vitamin D supplementation might be an additional therapeutical approach . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . P09038 promotes neurogenesis and neuroprotection and prolongs survival in a transgenic mouse model of Huntington 's disease . There is no satisfactory treatment for Huntington 's disease ( HD ) , a hereditary neurodegenerative disorder that produces chorea , dementia , and death . One potential treatment strategy involves the replacement of dead neurons by stimulating the proliferation of endogenous neuronal precursors ( neurogenesis ) and their migration into damaged regions of the brain . Because growth factors are neuroprotective in some settings and can also stimulate neurogenesis , we treated HD transgenic R6/2 mice from 8 weeks of age until death by s.c. administration of P09038 . P09038 increased the number of proliferating cells in the subventricular zone by approximately 30 % in wild-type mice , and by approximately 150 % in HD transgenic R6/2 mice . P09038 also induced the recruitment of new neurons from the subventricular zone into the neostriatum and cerebral cortex of HD transgenic R6/2 mice . In the striatum , these neurons were Q9UD71 -expressing medium spiny neurons , consistent with the phenotype of neurons lost in HD . P09038 was neuroprotective as well , because it blocked cell death induced by mutant expanded Htt in primary striatal cultures . P09038 also reduced polyglutamine aggregates , improved motor performance , and extended lifespan by approximately 20 % . We conclude that P09038 improves neurological deficits and longevity in a transgenic mouse model of HD , and that its neuroprotective and neuroproliferative effects may contribute to this improvement . Effects of chronic AVP P30518 blockade in congestive heart failure in sheep . Comparison with chronic P12821 inhibition . [ Association and interaction of AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 and P35354 genes on the risk of hypertension in Antioquian population ] . INTRODUCTION : Hypertension is a multifactorial disease influenced by genetic and environmental components , with its prevalence varying across ethnic groups . Manifold studies on blood pressure regulatory system genes have been carried out -such as the renin-angiotensin-aldosterone system , the sympathetic nervous system , endothelial factor , and sodium balance- , but the results yielded were inconsistent among populations . OBJECTIVES : To evaluate the effect of both variants in genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 P35354 , and the result of the individual ancestry component on hypertension and blood pressure levels among population in Antioquia . METHODS AND MATERIALS : 107 cases and 253 controls were genotyped for 12 variants on genes AGT , P30556 , P12821 , P07550 , P21728 , P35611 , P35612 , P20020 , P21731 y P35354 , and for 20 ancestry informative markers . The association of polymorphisms and their interactions , and the association of ancestral genetic composition with hypertension and blood pressure levels were examined . RESULTS : Genes P35612 , rs4852706 ( OR=3.0 ; p=0.023 ) ; P21728 , rs686 ( OR=0.38 ; p=0.012 ) and P07550 , rs1042718 ( OR=10.0 ; p=0.008 ) ; as well as genotypic combinations of P21728 and P30556 ; AGT and P35611 ; and P35611 to P20020 and P35354 were associated to hypertension . The Amerindian ancestry component was associated to some decrease in diastolic blood pressure . CONCLUSION : Variants on genes P35612 , P21728 , P07550 , P30556 , AGT , P35611 , P20020 and P35354 individually or interacting , are associated to hypertension . The Amerindian ancestry component has an effect on blood pressure . Genetics of idiopathic disseminated bronchiectasis . Bronchiectasis is an abnormal dilation of bronchi , consequent to the destruction of their walls . It is included in the category of obstructive pulmonary diseases , along with chronic obstructive pulmonary disease ( P48444 ) , asthma , and cystic fibrosis . In approximately 50 % of cases , bronchiectasis is associated with underlying conditions ; in the remainder , known causes are not ascertainable ( idiopathic bronchiectasis ) . A search for genetic determinants of this phenotype , with the cystic fibrosis gene as a candidate , has been performed by three independent groups . The results of this search agreed on the association of bronchiectasis with cystic fibrosis gene mutations and polymorphisms . The cystic fibrosis gene is also associated with bronchiectasis due to rheumatoid arthritis and allergic bronchopulmonary aspergillosis . A few other genes have been investigated in idiopathic bronchiectasis , with negative results . Idiopathic bronchiectasis is , therefore , to be considered as an obstructive multifactorial disorder belonging to the category of cystic fibrosis monosymptomatic diseases ( or P13569 -opathies ) , whose pathogenesis is influenced by environmental factors and other undetermined genes . Catalog of 178 variations in the Japanese population among eight human genes encoding G protein-coupled receptors ( GPCRs ) . We screened DNAs from 48 Japanese individuals for single-nucleotide polymorphisms ( SNPs ) in eight genes encoding G protein-coupled receptors ( GPCRs ) by directly sequencing the entire relevant genomic regions except for repetitive-sequence elements . This approach identified 147 SNPs and 31 insertion/deletion polymorphisms among the eight GPCR genes . On average , we identified one SNP in every 584 nucleotides . Of the 147 SNPs , 69 were identified in P30556 , 12 in P50052 , nine in P35414 , 20 in P37288 , nine in P30518 , 16 in P21728 , six in P08514 , and six in P43119 . Twenty-one SNPs were located in 5' flanking regions , 76 in introns , 32 in exons , and 18 in 3' flanking regions . These variants should contribute to investigations of possible correlations between genotypes and phenotypes as regards susceptibility to disease or responsiveness to drug therapy . The role of the subthalamic nucleus in DB01235 induced dyskinesia in 6-hydroxydopamine lesioned rats . DB01235 is the most effective treatment for Parkinson 's disease ( PD ) , but prolonged use leads to disabling motor complications including dyskinesia . Strong evidence supports a role of the subthalamic nucleus ( Q05639 ) in the pathophysiology of PD whereas its role in dyskinesia is a matter of controversy . Here , we investigated the involvement of Q05639 in dyskinesia , using single-unit extracellular recording , behavioural and molecular approaches in hemi-parkinsonian rats rendered dyskinetic by chronic DB01235 administration . Our results show that chronic DB01235 treatment does not modify the abnormal Q05639 activity induced by the 6-hydroxydopamine lesion of the nigrostriatal pathway in this model . Likewise , we observed a loss of Q05639 responsiveness to a single DB01235 dose both in lesioned and sham animals that received daily DB01235 treatment . We did not find any correlation between the abnormal involuntary movement ( AIM ) scores and the electrophysiological parameters of Q05639 neurons recorded 24 h or 20-120 min after the last DB01235 injection , except for the axial subscores . Nonetheless , unilateral chemical ablation of the Q05639 with ibotenic acid resulted in a reduction in global AIM scores and peak-severity of dyskinesia . In addition , Q05639 lesion decreased the anti-dyskinetogenic effect of buspirone in a reciprocal manner . Striatal protein expression was altered in dyskinetic animals with increases in ΔFosB , phosphoDARPP-32 , dopamine receptor ( DR ) D3 and P14416 / P21728 ratio . The Q05639 lesion attenuated the striatal molecular changes and normalized the P14416 / P21728 ratio . Taken together , our results show that the Q05639 plays a role , if modest , in the physiopathology of dyskinesias . Association of genetic polymorphisms with personality profile in individuals without psychiatric disorders . OBJECTIVE : Population-based twin studies demonstrate that approximately 40-50 % of the variability in personality dimensions results from genetic factors . This study assessed selected polymorphisms in the P21964 Val158Met , P21397 3'VNTR , 5HTTLPR , 102T/C 5- Q13049 , Q01959 3'VNTR and P14416 exon 8 genes and evaluated their association with personality profiles , anxiety levels , and depressiveness in healthy subjects . METHODS : This study included 406 unrelated ( mean age 38.51 years ) , mentally and somatically healthy Caucasian subjects of Polish origin . The prevalence of the gene variants mentioned above and their association with personality profiles , anxiety levels , and depressiveness was assessed using the Temperament and Character Inventory , NEO Five-Factor Inventory , Spielberger 's State-Trait Anxiety Inventory and Beck 's Depression Inventory . RESULTS : The effects of the 5HTTLPR gene on the s/s genotype and empathy ( P06681 ) were lowest in the entire group . The effects of gender , age and the Q13049 gene for the T/T genotype and attachment ( Q7Z3Z2 ) were highest in women . The effects of gender , age and the Q01959 gene on the 9/9 Q01959 genotype , compassion ( C4 ) and cooperativeness ( C ) were lowest in women . The effects of gender , age and the P21964 gene on the DB00134 / DB00134 genotype and neuroticism ( P04626 ) NEO-FFI were also lowest in women . CONCLUSIONS : Our results suggest considerable influence of individual genes on the formation of personality traits . The immunological effects of electrolyzed reduced water on the Echinostoma hortense infection in C57BL/6 mice . Electrolyzed reduced water ( ERW ) is widely used for drinking by people in Asia . The purpose of this study was to examine the immunological effect of ERW on the immunity of animals by supplying ERW to C57BL/6 mice infected with Echinostoma hortense metacercariae . In the non-infected groups , interleukin ( IL ) -4 ( p < 0.001 ) , P05113 , P22301 , IL-1beta , tumor necrosis factor ( P01375 ) -alpha and immunoglobulin ( Ig ) A expression of the group fed ERW ( ERW group ) increased in small intestine compared with the normal control group . In the case of infected groups , the group fed ERW ( ERW+E. hortense group ) showed the result that P05112 , P05113 , P22301 and Ig A expression increased , but IL-1beta and P01375 ( p < 0.001 ) decreased , and the number of goblet cells ( p < 0.001 ) and helix pomatia agglutinin ( Q9Y251 ) positive cells increased compared with the group without feeding ERW . However , adult worm recovery rate was markedly increased ( p < 0.05 ) . On the other hand , the expression of all the cytokines except P22301 in spleen was mildly increased but not significant statistically , and there was no significant difference in the numerical changes of white blood cell ( WBC ) . These results indicate that feeding ERW may have influence on the local immune response ( Th-1 type cytokines such as IL-1beta , P01375 ) in the small intestine but not on the systemic immune response . Dopamine selectively reduces GABA(B) transmission onto dopaminergic neurones by an unconventional presynaptic action . The functioning of midbrain dopaminergic neurones is closely involved in mental processes and movement . In particular the modulation of the inhibitory inputs on these cells might be crucial in controlling firing activity and dopamine ( DA ) release in the brain . Here , we report a concentration-dependent depressant action of dopamine on the GABA(B) IPSPs intracellularly recorded from dopaminergic neurones . Such effect was observed in spite of the presence of D(1)/ P14416 antagonists . A reduction of the GABA(B) IPSPs was also caused by noradrenaline ( norepinephrine ) and by L-beta-3,4-dihydroxyphenylalanine ( DB01235 ) , which is metabolically transformed into DA . The DA-induced depression of the IPSPs was partially antagonised by the alpha2 antagonists yohimbine and phentolamine . DA did not change the postsynaptic effects of the GABA(B) agonist baclofen , suggesting a presynaptic site of action . Furthermore , DA did not modulate the GABA(A)-mediated IPSP . The DA-induced depression of the GABA(B) IPSP occluded the depression produced by serotonin and was not antagonized by serotonin antagonists . The DA- and 5-HT-induced depression of the GABA(B) IPSP persisted when calcium and potassium currents were reduced in to the presynaptic terminals . These results describe an unconventional presynaptic , D(1) and D(2) independent action of DA on the GABA(B) IPSP . This might have a principal role in determining therapeutic/side effects of DB01235 and antipsychotics and could be also involved in drug abuse . [ Activation of coagulation cascade in children during an idiopathic nephrotic syndrome relapse ] . The objective of this study was to assess concentrations of selected markers of coagulation in children with relapse of idiopathic nephrotic syndrome during a 6-week therapy . Study groups : 22 subjects ( 32 relapses ) -- 14 males , 8 females ( mean age 7.15 +/- 1.5 y. ) with no thrombotic complications were included into the study . All children were clinically steroid-sensitive . METHODS : Coagulation markers ( platelet count , thrombin time , APTT , INR , fibrinogen 1 + 2 fragments ( F1 + 2 ) , thrombin-antithrombin complexes ( TAT ) , serum levels of D-dimer ( DD ) , fibrin monomers ( FM ) and antithrombin activity ( P01008 ) ) were measured three times : on admission , after 2 and 6 weeks . The control group consisted of 13 healthy children . RESULTS : Serum concentration of TAT or F1 + 2 did not differ between 3 stages ( p > 0.05 ) . However , values at 0 and 2 weeks were significantly higher than in control group ( p < 0.05 ) . We found no correlation between TAT or F1 + 2 and FBG , ALB , TCH , TG levels . [ table : see text ] CONCLUSIONS : The coagulation cascade in relapse of NS was activated during first 6 weeks of therapy whereas metabolic disturbances ( low ALB , high P02675 , TCH , TG , high platelets ) normalized . It is speculative whether it was caused by active immunological process but definitely it resulted in " prothrombotic state " in P01308 patients . Molecular genetics of bipolar disorder . Bipolar disorder ( BPD ) is an often devastating illness characterized by extreme mood dysregulation . Although family , twin and adoption studies consistently indicate a strong genetic component , specific genes that contribute to the illness remain unclear . This study gives an overview of linkage studies of BPD , concluding that the regions with the best evidence for linkage include areas on chromosomes 2p , 4p , 4q , 6q , 8q , 11p , 12q , 13q , 16p , 16q , 18p , 18q , 21q , 22q and Xq . Association studies are summarized , which support a possible role for numerous candidate genes in BPD including P21964 , Q01959 , Q13639 , P21917 , P14416 , P28223 , 5-HTT , the P59103 /G30 complex , Q9NRI5 , Q99572 , P21397 and P23560 . Animal models related to bipolar illness are also reviewed , with special attention paid to those with clear genetic implications . We conclude with suggestions for strategies that may help clarify the genetic bases of this complex illness . Human chromosome 3 and pig chromosome 13 show complete synteny conservation but extensive gene-order differences . A comparative map of human chromosome 3 ( HSA 3 ) and pig chromosome 13 ( SSC 13 ) was constructed using physically assigned pig sequence-tagged sites ( STSs ) . Pig STSs representing 11 HSA 3 genes , including v- P04049 murine leukemia viral oncogene homolog 1 ( P04049 ) , retinoic acid beta receptor ( P10826 ) , cholecystokinin ( CCK ) , pituitary transcription factor 1 ( P28069 ) , ceruloplasmin ( CP ) , guanine nucleotide binding protein , alpha-inhibiting polypeptide 2 ( P04899 ) , sucrase-isomaltase ( SI ) , rhodopsin ( P08100 ) , dopamine receptor D3 ( P35462 ) , growth-associated protein 43 ( P17677 ) , and somatostatin ( P61278 ) , were developed . Ten pig STSs were regionally mapped using a somatic cell hybrid panel ( SCHP ) to SSC 13 with 80-100 % concordance . Large-insert probes were obtained by screening a pig yeast artificial chromosome ( YAC ) library with primers for each STS . Several YACs were identified for P35462 , P17677 , P28069 , P08100 , SI , and P61278 for fluorescence in situ hybridization ( Q5TCZ1 ) mapping . Single gene and bi-color Q5TCZ1 with each pairwise combination were used to further define the gene order on SSC 13 . While these data confirm chromosome painting results showing that HSA 3 probes hybridize to a major portion of SSC 13 , they also demonstrate extensive gene-order differences between man and pig within this large conserved synteny group . Interestingly , several conserved chromosomal regions have been detected between pig and mouse that are not conserved between man and mouse , suggesting that the SSC 13 gene arrangement may be the closest to that of the ancestral eutherian chromosome . Task-dependent interactions between dopamine D2 receptor polymorphisms and DB01235 in patients with Parkinson 's disease . Variants in genes regulating dopamine transmission affect performance on tasks including working memory and executive function as well as temporal processing and sequence learning . In the current study , we determined whether a dopamine D2 receptor DNA sequence polymorphism interacts with DB01235 during motor tasks in patients with Parkinson 's disease ( PD ) . Forty-five PD patients were genotyped for the P14416 polymorphism ( rs 1076560 , G > T ) . Patients performed an explicit motor sequence learning task and the grooved pegboard test in both ON and OFF DB01235 states . For motor sequence learning , P14416 genotype mediated DB01235 effects such that DB01235 associated improvements were only observed in the minor T allele carriers ( associated with lower D2 receptor availability , t10=-2.71 , p=0.022 ) , whereas G homozygotes showed no performance change with DB01235 . For the grooved pegboard test , performance improved with DB01235 independent of patients ' P14416 genotype . Collectively these results demonstrate that common P14416 allelic differences found in the human population may explain how dopamine differentially contributes to performance across tasks and individuals . The low-potency , voltage-dependent Q12809 blocker propafenone -- molecular determinants and drug trapping . The molecular determinants of high-affinity human ether-a-go-go-related gene ( Q12809 ) potassium channel blockade by methanesulfonanilides include two aromatic residues ( Phe656 and Tyr652 ) on the inner helices ( S6 ) and residues on the pore helices that face into the inner cavity , but determinants for lower-affinity Q12809 blockers may be different . In this study , alanine-substituted Q12809 channel mutants of inner cavity residues were expressed in Xenopus laevis oocytes and were used to characterize the Q12809 channel binding site of the antiarrhythmic propafenone . DB01182 's blockade of Q12809 was strongly dependent on residue Phe656 but was insensitive or weakly sensitive to mutation of Tyr652 , Thr623 , Ser624 , Val625 , Gly648 , or Val659 and did not require functional inactivation . Homology models of Q12809 based on KcsA and MthK crystal structures , representing the closed and open forms of the channel , respectively , suggest propafenone is trapped in the inner cavity and is unable to interact exclusively with Phe656 in the closed state ( whereas exclusive interactions between propafenone and Phe656 are found in the open-channel model ) . These findings are supported by very slow recovery of wild-type Q12809 channels from block at -120 mV , but extremely rapid recovery of D540K channels that reopen at this potential . The experiments and modeling suggest that the open-state propafenone binding-site may be formed by the Phe656 residues alone . The binding site for propafenone ( which may involve pi-stacking interactions with two or more Phe656 side-chains ) is either perturbed or becomes less accessible because of closed-channel gating . This provides further evidence for the existence of gating-induced changes in the spatial location of Phe656 side chains . Very early-onset lone atrial fibrillation patients have a high prevalence of rare variants in genes previously associated with atrial fibrillation . BACKGROUND : Atrial fibrillation ( AF ) is the most common cardiac arrhythmia . Currently , 14 genes important for ion channel function , intercellular signaling , and homeostatic control have been associated with AF . OBJECTIVE : We hypothesized that rare genetic variants in genes previously associated with AF had a higher prevalence in early-onset lone AF patients than in the background population . METHODS : Sequencing results of P51787 , Q12809 , Q14524 , P22460 , Q9UK17 , P15382 , 2 , 5 , P63252 , P35498 -3B , P01160 , and P36382 from 192 early-onset lone AF patients were compared with data from the National Heart , Lung , and Blood Institute Exome Variant Server consisting of 6503 persons from 18 different cohort studies . RESULTS : Among the lone AF patients , 29 ( 7.6 % ) alleles harbored a novel or very rare variant ( minor allele frequency < 0.1 in the Exome Variant Server ) , a frequency that was significantly higher than what was found in the reference database ( 4.1 % ; with minor allele frequency < 0.1 ; P = .0012 ) . Previously published electrophysiological data showed that 96 % ( n = 23 ) of the rare variants that has been functionally investigated ( n = 24 ) displayed significant functional changes . CONCLUSIONS : We report a much higher prevalence of rare variants in genes associated with AF in early-onset lone AF patients than in the background population . By presenting these data , we believe that we are the first to provide quantitative evidence for the role of rare variants across AF susceptibility genes as a possible pathophysiological substrate for AF . Increased blood pressure and loss of anp-induced natriuresis in mice lacking Q9UD71 gene . Atrial natriuretic peptide ( P01160 ) is an important regulator of sodium metabolism and indirectly of blood pressure . Evidence has accumulated that P01160 regulates sodium metabolism through a cascade of steps involving an increase in the level of cGMP , activation of cGMP-dependent protein kinase ( PKG ) , and inhibition of renal tubular Na+ , K+-ATPase activity . One of the major substrates for PKG is Q9UD71 . In the present study we observed that P01160 does not induce natriuresis in mice that lack Q9UD71 . In contrast , there was a 4-fold increase in urinary sodium excretion following P01160 administration to wild type mice . P01160 as well as Zaprinast , a selective inhibitor of cGMP phosophodiesterase , inhibited renal Na+ , K+-ATPase activity in wild type mice but had no such effect in mice lacking Q9UD71 . Mean arterial blood pressure , measured in conscious animals , was significantly increased in Q9UD71 deficient mice as compared to wild type mice . The results confirm that Q9UD71 acts as a third messenger in the P01160 signaling pathway in renal tissue and suggest an important role of Q9UD71 in the maintenance of normal blood pressure . Molecular evolution of the oxytocin-oxytocin receptor system in eutherians . DB00107 ( P01178 ) is a nine-amino-acid peptide hormone that is mainly released at the times of uterine contractions during parturition and milk ejection during lactation , whereas a similar peptide hormone , arginine vasopressin , primarily exerts direct antidiuretic action on the kidney and causes vasoconstriction of the peripheral vessels . The genes coding for these peptides are tandemly located on the same chromosome . A tandem duplication occurring in the common ancestor of jawed vertebrates has been proposed as responsible . In contrast to the two peptide hormones , only one oxytocin receptor ( P30559 ) but three arginine vasopressin receptors ( P37288 , P47901 , and P30518 ) are known ; these receptors probably arose from two rounds of genome duplication in the common ancestor of vertebrates . In this study , we addressed the molecular evolution of the P01178 - P30559 system in eutherians . Our analyses suggest that an amino acid change from isoleucine to lysine on the eighth site ( I8L ) of the peptide , which corresponded to a change from mesotocin to P01178 , had occurred during the common ancestral lineage of eutherians . At around the same time that the emergence of P01178 occurred , functional constraints on the P01178 receptor ( pre- P30559 ) might have relaxed , and a series of nonsynonymous substitutions might have accumulated . Only a few of these nonsynonymous substitutions might have contributed to reestablishing the molecular relationship between the P01178 ligand and its receptor , after which functional constraints on the P30559 were reinstated . Since the P01178 - P30559 system plays an important role in eutherians , the evolution of the P01178 - P30559 system was probably an essential component of the genesis of the eutherian signature . Sustained increase of PKA activity in the postcommissural putamen of dyskinetic monkeys . Levodopa-induced dyskinesias ( LID ) are a frequent complication of Parkinson 's disease pharmacotherapy that causes significant disability and narrows the therapeutic window . Pharmacological management of LID is challenging partly because the precise molecular mechanisms are not completely understood . Here , our aim was to determine molecular changes that could unveil targetable mechanisms underlying this drug complication . We examined the expression and downstream activity of dopamine receptors ( DR ) in the striatum of 1-methyl-4-phenyl-1,2,3,6 tetrahydropiridine ( MPTP ) -lesioned monkeys with and without DB01235 treatment . Four monkeys were made dyskinetic and other four received a shorter course of DB01235 and did not develop LID . Our results show that DB01235 treatment induces an increase in P14416 and P35462 expression in the postcommissural putamen , but only P35462 is correlated with the severity of LID . Dyskinetic monkeys show a hyperactivation of the canonical P21728 -signaling pathway , measured by an increased phosphorylation of protein kinase A ( PKA ) and its substrates , particularly Q9UD71 . In contrast , activation of the P14416 -signaling pathway , visible in the levels of Akt phosphorylated on Thr308 and GSK3β on Ser9 , is associated with DB01235 treatment , independently of the presence of dyskinesias . Our data clearly demonstrate that dyskinetic monkeys present a dysregulation of the P35462 receptor and the P21728 pathway with a sustained increase of PKA activity in the postcommissural putamen . Importantly , we found that all signaling changes related to long-term DB01235 administration are exquisitely restricted to the postcommissural putamen , which may be related to the recurrent failure of pharmacological approaches . Maxi-anion channel as a candidate pathway for osmosensitive DB00171 release from mouse astrocytes in primary culture . In the present study , we aimed to evaluate the pathways contributing to DB00171 release from mouse astrocytes during hypoosmotic stress . We first examined the expression of mRNAs for proteins constituting possible DB00171 -releasing pathways that have been suggested over the past several years . In RT-PCR analysis using both control and osmotically swollen astrocytes , amplification of cDNA fragments of expected size was seen for connexins ( P08034 , P35212 , P17302 ) , pannexin 1 ( Px1 ) , the Q99572 receptor , MRP1 and P08183 , but not P13569 . Inhibitors of exocytotic vesicular release , gap junction hemi-channels , P13569 , MRP1 , P08183 , the Q99572 receptor , and volume-sensitive outwardly rectifying chloride channels had no significant effects on the massive DB00171 release from astrocytes . In contrast , the hypotonicity-induced DB00171 release from astrocytes was most effectively inhibited by gadolinium ( 50 muM ) , an inhibitor of the maxi-anion channel , which has recently been shown to serve as a pathway for DB00171 release from several other cell types . Thus , we propose that the maxi-anion channel constitutes a major pathway for swelling-induced DB00171 release from cultured mouse astrocytes as well . P21728 behavioral responsitivity following selective lesions of the striatal patch compartment during development . The behavioral effects of selective destruction of the dopamine ( DA ) input to the patch compartment of rat striatum early in development was investigated . Rat pups were given bilateral intrastriatal ( i.s. ) injections of the neurotoxin 6-hydroxydopamine ( 6-OHDA ) on day of birth ( P0 ) or postnatal day 1 ( P1 ) , which resulted in selective behavioral alterations following DA agonist treatment in adulthood . Neonatally-lesioned rats exhibited self-biting behavior following treatment with the DA precursor L-dihydroxyphenylalanine ( DB01235 ) . In response to treatment with the selective D1 agonist SKF38393 , there was an increased incidence of abnormal perioral movements . The cataleptogenic effects of the D1 antagonist SCH23390 and the D2 antagonist haloperidol were also studied . Neonatally-lesioned rats were significantly less cataleptic compared to control rats following D1 antagonist treatment , but not following D2 antagonist treatment . Autoradiographs of [3H]mazindol binding to DA uptake sites ( a measure of DA terminal density ) showed a ' patchy ' loss of approx . 40-50 % in striatal tissue sections derived from the i.s. lesioned rats . These data suggest that injections of 6-OHDA into the striatum during this early postnatal period cause a DA lesion that results in long-term effects on a D1 receptor system . The A1 allele of the human D2 dopamine receptor gene is associated with increased activity of striatal L-amino acid decarboxylase in healthy subjects . The A1 allele of the TaqI restriction fragment length polymorphism ( RFLP ) of the human dopamine D2 receptor gene ( P14416 ) is associated with a low density of D2 dopamine receptors in the striatum . Because of the important role of D2 autoreceptors in regulating dopamine synthesis , we aimed to examine whether subjects with the A1 allele have altered presynaptic dopamine function in the brain . We also studied the effects of two other P14416 polymorphisms , C957 T and -- 141C Ins/Del , which have been suggested to affect D2 receptor levels in brain . The relationships between the TaqIA RFLP , C957 T and -- 141C Ins/Del polymorphisms and striatal dopamine synthesis in 33 healthy Finnish volunteers were studied using positron emission tomography and [18F]fluorodopa ( [18F]FDOPA ) , a radiolabelled analog of the dopamine precursor DB01235 . Heterozygous carriers of the A1 allele ( A1/A2 ; 10 subjects ) had significantly higher ( 18 % ) [18F]FDOPA uptake in the putamen than subjects without the A1 allele ( A2/A2 ; 23 subjects ) . C957 T and -- 141C Ins/Del polymorphisms did not significantly affect [18F]FDOPA Ki values . These results demonstrate that the A1 allele of P14416 gene is associated with increased striatal activity of aromatic L-amino acid decarboxylase , the final enzyme in the biosynthesis of dopamine and the rate-limiting enzyme for trace amine ( e.g. beta-phenylethylamine ) synthesis . The finding can be explained by lower D2 receptor expression leading to decreased autoreceptor function , and suggests that dopamine and/or trace amine synthesis rate is increased in the brains of A1 allele carriers . DB01235 -treatment in primates disrupts the expression of A(2A) adenosine-CB(1) cannabinoid- P14416 heteromers in the caudate nucleus . The molecular basis of priming for DB01235 -induced dyskinesias in Parkinson 's disease ( PD ) , which depends on the indirect pathway of motor control , is not known . In rodents , the indirect pathway contains striatopallidal GABAergic neurons that express heterotrimers composed of A(2A) adenosine , CB(1) cannabinoid and D(2) dopamine receptors that regulate dopaminergic neurotransmission . The present study was designed to investigate the expression of these heteromers in the striatum of a primate model of Parkinson 's disease and to determine whether their expression and pharmacological properties are altered upon DB01235 treatment . By using the recently developed in situ proximity ligation assay and by identification of a biochemical fingerprint , we discovered a regional distribution of A(2A)/CB(1) /D(2) receptor heteromers that predicts differential D(2)-mediated neurotransmission in the caudate-putamen of Macaca fascicularis . Whereas heteromers were abundant in the caudate nucleus of both naïve and MPTP-treated monkeys , DB01235 treatment blunted the biochemical fingerprint and led to weak heteromer expression . These findings constitute the first evidence of altered receptor heteromer expression in pathological conditions and suggest that drugs targeting A(2A)-CB(1) -D(2) receptor heteromers may be successful to either normalize basal ganglia output or prevent DB01235 -induced side effects . Association of dopamine-related gene alleles , smoking behavior and decline in FEV1 in subjects with P48444 : findings from the lung health study . Cigarette smoking is the major risk factor for chronic obstructive pulmonary disease ( P48444 ) . Specific dopamine related gene alleles have previously been found to be associated with smoking initiation , maintenance and cessation . We investigated the association between specific dopamine related gene alleles and both change in smoking behavior and lung function change over time in individuals with mild-to-moderate P48444 . Subjects included a subset of participants in the Lung Health Study ( LHS ) , a smoking intervention study in smokers with mild to moderate P48444 . Smoking status was determined and lung function performed at baseline and annually for 5 years . In post-hoc analyses , we assessed the association of the dopamine receptor ( P14416 ) TaqI A1(+) allele ( A1A1 , A1A2 genotypes ) and A1(-) allele ( A2A2 genotype ) , and the dopamine transporter ( Q01959 ) 9R(+) allele ( 9R9R and 9R10R genotypes ) and 9R(-) allele ( 10R10R genotype ) with both changes in smoking status and lung function in a subset of LHS subjects . No significant associations were noted between variants in these genes and success in smoking cessation . However , in exploratory analyses that did not adjust for multiple comparisons , sustained male ( but not female ) quitters with the P14416 A1(-) allele and/or the Q01959 9R(+) allele showed an accelerated decline in Q99581 (1) similar to that of continuing smokers over 5 years after quitting smoking . These preliminary findings suggest that dopamine-related genes may play a role in the progression of P48444 , at least in the subset of male ex-smokers whose disease continues to progress despite sustained quitting , and warrants additional confirmatory and mechanistic studies . Activity of retinoic acid receptor-gamma selectively binding retinoids alone and in combination with interferon-gamma in breast cancer cell lines . Retinoids modulate several cell functions and especially inhibit the growth of a wide variety of cells including breast cancer . Retinoic acid receptor-gamma ( P13631 ) has been shown to mediate the antiproliferative activity of retinoids . To further test this hypothesis we examined the effects of different P13631 selectively binding retinoids ( CD2325 , CD2247 , CD666 and CD437 ) on breast cancer cell lines . With exception of CD2247 , all retinoids inhibited proliferation of MCF-7 , SKBR-3 , T47D and ZR-75-1 breast cancer cell lines , similar to the natural compound all-trans retinoic acid ( DB00755 ) . In addition , all 4 compounds were able to act synergistically with interferon-gamma ( P01579 ) in all breast cancer cell lines including the retinoid-resistant BT-20 and 734-B lines . In functional transactivation assays we demonstrated that only in the MCF-7 cell line , TPA-mediated AP-1 activity was suppressed only by DB00755 and CD2325 , whereas in SKBR-3 , another RA-sensitive breast cancer cell line , it was not . The synergistic antiproliferative activity involving retinoids and P01579 could not be explained by an enhanced anti-AP-1 activity . No correlation was found between expression of RARs and cellular retinoic acid binding proteins ( CRABPs ) and antiproliferative effects of the retinoids . P13631 selectively binding retinoids are potent inhibitors of breast cancer cell proliferation , alone and in combination with P01579 . For this reason and because of a possible low toxicity , as compared with retinoic acid , we speculate that these P13631 selective binding retinoids might be of clinical importance . Readjusting the localization of long QT syndrome gene on chromosome 11p15 . Long QT syndrome ( LQT ) is an autosomal dominant cardiac disease characterized by ventricular arrhythmia . A first locus for LQT has been identified on chromosome 11p15.5 ( LQT1 ) , closely linked to P01112 . To refine the location of LQT1 , microsatellites were genotyped in 8 French families and the following order was determined : tel- P01112 - P21917 -D11S922-D11S4046- P01344 - P01308 -TH-D11S1318-D11S1323-D11S1338-D11S90 9-D11S1346-cen . By haplotype analysis , 12 crossing-over events were identified in affected and unaffected subjects , delineating the LQT1 candidate region to 7 cM . This new delineated localization between D11S1318 and D11S1323 is in a more centromeric region than previously thought and is 5 cM proximal to P01112 . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Integrative gene network analysis provides novel regulatory relationships , genetic contributions and susceptible targets in autism spectrum disorders . Autism spectrum disorders ( ASDs ) are a group of diseases exhibiting impairment in social drive , communication/language skills and stereotyped behaviors . Though an increased number of candidate genes and molecular interactions have been identified by various approaches , the pathogenesis remains elusive . Based on clinical observations , data from accessible GWAS and expression datasets we identified ASDs gene candidates . Integrative gene network and a novel CNV-centric Node Network ( CNN ) analysis method highlighted ASDs-associated key elements and biological processes . Functional analysis identified neurological functions including synaptic cholinergic receptor ( CHRNA ) families , dopamine receptor ( P14416 ) , and correlations between social behavior and oxytocin related pathways . CNN analysis of genome-wide genetic and expression data identified inheritance-related clusters related to P60484 / Q92574 / Q06787 and mTor/PI3K regulation . Integrative analysis identified potential regulators of networks , specifically P01375 and beta-estradiol , suggesting a potential central role in ASDs . Our data provide information on potential disease mechanisms , and key regulators that may generate novel postulations , and diagnostic molecular biomarkers . Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . Dopaminergic , serotonergic , and oxytonergic candidate genes associated with infant attachment security and disorganization ? In search of main and interaction effects . BACKGROUND AND METHODS : In two birth cohort studies with genetic , sensitive parenting , and attachment data of more than 1,000 infants in total , we tested main and interaction effects of candidate genes involved in the dopamine , serotonin , and oxytocin systems ( P21917 , P14416 , P21964 , 5-HTT , P30559 ) on attachment security and disorganization . Parenting was assessed using observational rating scales for parental sensitivity ( Ainsworth , Bell , & Stayton , 1974 ) , and infant attachment was assessed with the Strange Situation Procedure . RESULTS : We found no consistent additive genetic associations for attachment security and attachment disorganization . However , specific tests revealed evidence for a codominant risk model for P21964 Val158Met , consistent across both samples . Children with the DB00161 / DB00134 genotype showed higher disorganization scores ( combined effect size d = .22 , CI = .10-.34 , p < .001 ) . Gene-by-environment interaction effects were not replicable across the two samples . CONCLUSIONS : This unexpected finding might be explained by a broader range of plasticity in heterozygotes , which may increase susceptibility to environmental influences or to dysregulation of emotional arousal . This study is unique in combining the two largest attachment cohorts with molecular genetic and observed rearing environment data to date . Dopamine receptor D4 polymorphism predicts the effect of DB01235 on gambling behavior . BACKGROUND : There is ample evidence that a subgroup of Parkinson 's disease patients who are treated with dopaminergic drugs develop certain behavioral addictions such as pathological gambling . The fact that only a subgroup of these patients develops pathological gambling suggests an interaction between dopaminergic drug treatment and individual susceptibility factors . These are potentially of genetic origin , since research in healthy subjects suggests that vulnerability for pathological gambling may be linked to variation in the dopamine receptor D4 ( P21917 ) gene . Using a pharmacogenetic approach , we investigated how variation in this gene modulates the impact of dopaminergic stimulation on gambling behavior in healthy subjects . METHODS : We administered 300 mg of L-dihydroxyphenylalanine ( DB01235 ) or placebo to 200 healthy male subjects who were all genotyped for their P21917 polymorphism . Subjects played a gambling task 60 minutes after DB01235 administration . RESULTS : Without considering genetic information , DB01235 administration did not lead to an increase in gambling propensity compared with placebo . As expected , however , an individual 's P21917 polymorphism accounted for variation in gambling behavior after the administration of DB01235 . Subjects who carry at least one copy of the 7-repeat allele showed an increased gambling propensity after dopaminergic stimulation . CONCLUSIONS : These findings demonstrate that genetic variation in the P21917 gene determines an individual 's gambling behavior in response to a dopaminergic drug challenge . They may have implications for the treatment of Parkinson 's disease patients by offering a genotype approach for determining individual susceptibilities for pathological gambling and may also afford insights into the vulnerability mechanisms underlying addictive behavior . Anti-Parkinson 's disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson 's disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson 's disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson 's disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene/drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 /benztropine , P21964 /levodopa and entacapone , P20813 /selegiline , P22309 /entacapone , P14416 /ropinirole , pramipexole and cabergoline , and P35462 /ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson 's disease drug response includes P05177 in the response to ropinirole and rasagiline , and P08684 in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson 's disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson 's disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests . Blockade of globus pallidus adenosine A(2A) receptors displays antiparkinsonian activity in 6-hydroxydopamine-lesioned rats treated with D(1) or P14416 agonists . We have recently demonstrated how antagonism of adenosine A(2A) receptors within the globus pallidus ( GP ) ipsilateral to dopaminergic denervation potentiates contralateral rotational behavior induced by the dopamine precursor DB01235 in 6-hydroxydopamine-lesioned hemiparkinsonian rats . To further characterize the influence of pallidal A(2A) receptor blockade on the motor stimulant effects elicited by dopamine receptor activation , hemiparkinsonian rats were infused with the water-soluble A(2A) antagonist P35240 BT2 in the GP , alone or in combination with systemic administration of either SKF 38393 or quinpirole , to stimulate dopamine D(1) or D(2) receptors , respectively . P35240 BT2 alone ( 5 mug/1 mul ) neither altered motor behavior nor produced postural asymmetry . In contrast , the contralateral rotations elicited by SKF 38393 ( 1.5 mg/kg ) as well as quinpirole ( 0.05 mg/kg ) were potentiated by the concomitant intrapallidal infusion of P35240 BT2 . The results of this study demonstrate that blockade of pallidal A(2A) receptors exerts a facilitatory influence on the motor effects produced by the selective stimulation of either D(1) or D(2) dopamine receptors in hemiparkinsonian rats and suggest an involvement of GP in the antiparkinsonian activity of A(2A) receptor antagonists . P14416 desensitization by dopamine or corticotropin releasing factor in ventral tegmental area neurons is associated with increased glutamate release . Neurons of the ventral tegmental area ( VTA ) are the source of dopaminergic ( DAergic ) input to important brain regions related to addiction . Prolonged exposure of these VTA neurons to moderate concentrations of dopamine ( DA ) causes a time-dependent decrease in DA-induced inhibition , a complex desensitization called DA inhibition reversal ( P30518 ) . P30518 is mediated by conventional protein kinase C ( cPKC ) through concurrent stimulation of D2 and D1-like DA receptors , or by D2 stimulation concurrent with activation of some Gq-linked receptors . DB01285 releasing factor ( CRF ) acts via Gq , and can modulate glutamater neurotransmission in the VTA . In the present study , we used brain slice electrophysiology to characterize the interaction of DA , glutamate antagonists , and CRF agonists in the induction and maintenance of P30518 in the VTA . Glutamate receptor antagonists blocked induction but not maintenance of P30518 . Putative blockers of neurotransmitter release and store-operated calcium channels blocked and reversed P30518 . CRF and the CRF agonist urocortin reversed inhibition produced by the D2 agonist quinpirole , consistent with our earlier work indicating that Gq activation reverses quinpirole-mediated inhibition . In whole cell recordings , the combination of urocortin and quinpirole , but not either agent alone , increased spontaneous excitatory postsynaptic currents ( sEPSCs ) in VTA neurons . Likewise , the combination of a D1-like receptor agonist and quinpirole , but not either agent alone , increased sEPSCs in VTA neurons . In summary , desensitization of D2 receptors induced by dopamine or CRF on DAergic VTA neurons is associated with increased glutamatergic signaling in the VTA . DB01184 treatment for gastroparesis : demographic and pharmacogenetic characterization of clinical efficacy and side-effects . BACKGROUND : DB01184 is a useful alternative to metoclopramide for treatment of gastroparesis due to better tolerability . Effectiveness and side-effects from domperidone may be influenced by patient-related factors including polymorphisms in genes encoding drug-metabolizing enzymes , drug transporters , and domperidone targets . AIMS : The aim of this study was to determine if demographic and pharmacogenetic parameters of patients receiving domperidone are associated with response to treatment or side-effects . METHODS : Patients treated with domperidone for gastroparesis provided saliva samples from which DNA was extracted . Fourteen single-nucleotide polymorphisms ( SNPs ) in seven candidate genes ( P08183 , P10635 , P14416 , P15382 , Q9Y6J6 , Q12809 , P51787 ) were used for genotyping . SNP microarrays were used to assess single-nucleotide polymorphisms in the ADRA1A , P35368 , and P25100 loci . RESULTS : Forty-eight patients treated with domperidone participated in the study . DNA was successfully obtained from each patient . Age was associated with effectiveness of domperidone ( p=0.0088 ) . Genetic polymorphism in Q12809 was associated with effectiveness of domperidone ( p=0.041 ) . The efficacious dose was associated with polymorphism in P08183 gene ( p=0.0277 ) . The side-effects of domperidone were significantly associated with the SNPs in the promoter region of P25100 gene . CONCLUSIONS : Genetic characteristics associated with response to domperidone therapy included polymorphisms in the drug transporter gene P08183 , the potassium channel Q12809 gene , and α1D -- adrenoceptor P25100 gene . Age was associated with a beneficial response to domperidone . If verified in a larger population , this information might be used to help determine which patients with gastroparesis might respond to domperidone and avoid treatment in those who might develop side-effects . Fibroblast growth factor-2 in hyperplastic pituitaries of D2R knockout female mice . P14416 ( D2R ) knockout ( KO ) female mice develop chronic hyperprolactinemia and pituitary hyperplasia . Our objective was to study the expression of the mitogen fibroblast growth factor ( P09038 ) and its receptor , P11362 , comparatively in pituitaries from KO and wild-type ( WT ) female mice . We also evaluated P09038 subcellular localization and P09038 effects on pituitary function . P09038 -induced prolactin release showed a similar response pattern in both genotypes , even though basal and P09038 -stimulated release was higher in KO . P09038 stimulated pituitary cellular proliferation ( MTS assay and [(3)H]thymidine incorporation ) , with no differences between genotypes . P09038 concentration ( measured by ELISA ) in whole pituitaries or cultured cells was lower in KO ( P < 0.00001 and 0.00014 ) . Immunofluorescence histochemistry showed less P09038 in pituitaries from KO females and revealed a distinct P09038 localization pattern between genotypes , being predominantly nuclear in KO and cytosolic in WT pituitaries . Finally , P09038 could not be detected in the conditioned media from pituitary cultures of both genotypes . P11362 levels ( Western blot and immunohistochemistry ) were higher in pituitaries of KO . Basal concentration of phosphorylated ERKs was lower in KO cells ( P = 0.018 ) . However , when stimulated with P09038 , a significantly higher increment of P29323 phosphorylation was evidenced in KO cells ( P < or = 0.02 ) . We conclude that disruption of the D2R caused an overall decrease in pituitary P09038 levels , with an increased distribution in the nucleus , and increased P11362 levels . These results are important in the search for reliable prognostic indicators for patients with pituitary dopamine-resistant prolactinomas , which will make tumor-specific therapy possible . Radiation hybrid map of 13 loci on the long arm of chromosome 5 . Radiation hybrid mapping was used in conjunction with a natural deletion mapping panel to predict the order of and distance between 13 loci in the distal portion of the long arm of human chromosome 5 . A panel of irradiation hybrids containing fragments of 5q was generated from an P00492 + Chinese hamster-human cell hybrid containing a derivative chromosome 5 [ der(5)t(4;5) ( 5qter ---- 5p15.1::4p15.1 ---- 4pter ) ] as its only human DNA . One hundred nine radiation hybrids containing human DNA were screened with polymerase chain reaction primer sets representing nine genes encoding growth factors , growth factor receptors , or hormone receptors ( P08700 , P05112 , P05113 , P07333 , P05230 , P07550 , GRL , P14867 , and P21728 ) as well as four other loci ( P16591 , P09486 , P62263 , and P08571 ) to generate a radiation hybrid map of the area 5q21-q35 . A physical map predicting the order of and distance between the 13 loci was constructed based on segregation of the 13 loci in hybrid clones . The radiation hybrid panel will be useful as a mapping tool for determining the location and order of other genes and polymorphic loci in this region as well as for generating new DNA probes from specific regions . Dopamine receptor agonists reverse behavioral abnormalities of alpha-synuclein transgenic mouse , a new model of Parkinson 's disease . Parkinson 's disease ( PD ) is characterized by loss of nigral dopaminergic ( DAergic ) neurons and presence of Lewy bodies , whose major component is alpha-synuclein . We had previously generated transgenic mice termed Syn130m that express truncated human alpha-synuclein ( amino acid residues 1-130 ) in DAergic neurons . Syn130m mice showed significant loss of DAergic neurons in the substantia nigra pars compacta . Subsequently , the striatal DA level and spontaneous locomotor activity of the mice were decreased significantly . In the present study , we investigated behavioral responses of Syn130m mice to DB01235 and DA receptor agonists . Administration of DB01235 dose dependently ameliorated the reduction of spontaneous locomotor activity of Syn130m mice . Similarly , D(2) agonists , quinpirole and talipexole , and a D1/D2 agonist , pergolide , were effective against the reduction . Syn130m mice also showed significant reduction in exploratory behavior compared with non-Tg littermates when they were placed in a novel environment , but this abnormality was ameliorated by treatment with pergolide . These results strongly suggest that the behavioral abnormalities of Syn130m mice were caused by low striatal DA content . On the other hand , the expression of postsynaptic D(2)-like receptors ( P14416 ) in the striatum was not increased in Syn130m mice , although the low striatal DA level is known to induce compensatory expression of P14416 . Because the abnormalities could be rectified by treatment with DA receptor agonists , it is likely that Syn130m mice provide a useful tool to explore therapeutic possibilities for PD as a new animal model of the disease . Therapy with interferon-beta modulates endogenous catecholamines in lymphocytes of patients with multiple sclerosis . OBJECTIVE : To investigate the endogenous dopaminergic/adrenergic system of lymphocytes in multiple sclerosis ( MS ) patients during treatment with interferon ( IFN ) -beta . METHODS : Patients with relapsing-remitting MS undergoing IFN-beta treatment were prospectively studied during the first year of treatment . Circulating lymphocytes were obtained at baseline and after 1 , 3 , 6 and 12 months of treatment and assayed for catecholamine ( CA ) production and mRNA expression of tyrosine hydroxylase ( TH , the rate-limiting enzyme in the synthesis of CA ) , beta(2)-adrenoceptors ( AR ) and D2 , D3 and D5 dopaminergic receptors ( DR ) . RESULTS : In cells from patients treated with IFN-beta for 12 months the production of CA hugely increased and was less sensitive to P01579 -induced inhibition . Expression of mRNA for TH , beta(2)-AR and P21918 was already enhanced after 1 month and further increased up to 6-12 months of treatment . On the contrary , P14416 mRNA progressively decreased and P35462 mRNA did not significantly change over the whole study period . CONCLUSIONS : In MS patients IFN-beta treatment enhances the ability of lymphocytes to produce CA , and induces extensive modifications of both beta(2)-AR and DR-operated pathways . The clinical relevance of these effects deserves consideration . Nonlinkage of bipolar illness to tyrosine hydroxylase , tyrosinase , and D2 and D4 dopamine receptor genes on chromosome 11 . OBJECTIVE : Previous linkage and allelic association studies using DNA polymorphisms , cosegregation of cytogenetic abnormalities with psychiatric illness , and assignment of genes involved in neutotransmitter metabolism suggested that chromosome 11 may harbor a gene predisposing to bipolar illness . The authors examined linkage in the families of 14 probands with bipolar illness , with the candidate genes tyrosine hydroxylase ( TH ) , D4 dopamine receptor ( P21917 ) at 11p15 , tyrosinase ( P14679 ) at 11q14-q21 , and D2 dopamine receptor ( P14416 ) at 11q22-q23 , as well as with the c-Harvey-ras oncogene ( P01112 ) and insulin gene ( P01308 ) , both located at 11p15 , a region that previously showed linkage to bipolar illness . METHOD : The genetic data were analyzed with both lod score analysis ( parametric ) and affected-sib-pair analysis ( nonparametric ) ; both narrow and broad definitions of the clinical phenotype were used . Further influences of diagnostic uncertainties were accounted for by using diagnostic probability classes weighing the stability of each phenotype . RESULTS : Two-point linkage results excluded close linkage of bipolar illness to each candidate gene ; negative results were also obtained when the narrow definition of the clinical phenotype was used . Moreover , multipoint linkage analysis of P01112 and P01308 excluded the 11p15 region encompassing both P21917 and TH . In agreement with the negative linkage results , affected-sib-pair analysis did not show preferential sharing of marker alleles at any of the candidate genes . CONCLUSIONS : The negative results obtained under different genetic models exclude a frequent role for P21917 , TH , P14679 , and P14416 in the pathogenesis of bipolar illness . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . AP-2beta represses P21728 gene transcription in neuro2a cells . Expression of the P21728 in brain is restricted to specific neuronal populations . To investigate the mechanism of this selective expression , we localized a silencer upstream of the human D(1A) gene and identified its binding transcription factor in the D(1A)-negative neural cell line Neuro2a . Using deletion CAT analysis , we narrowed this silencer to the region between nucleotides -561 and -532 relative to the CAP site . This 30-bp region , designated D1AS1 , contains a sequence homologous to the P05549 binding site and binds to a factor that also interacts with the P05549 consensus sequence . In gel supershift assays , this factor is recognized by anti-AP-2beta antibody . Co-transfection of Neuro2a cells with an AP-2beta expression vector repressed the basal CAT activity of D(1A) promoter-reporter plasmids in a D1AS1-dependent manner . RT-PCR analysis indicated that , among P05549 family members , Neuro2a cells express only AP-2beta . Furthermore , co-transfection of these cells with decoy oligonucleotides corresponding to the D1AS1 sequence de-repressed the D(1A) gene promoter . Unlike in Neuro2a cells , AP-2beta could not repress the D(1A) promoter in the D(1A)-positive neural cell line , NS20Y . In addition , the expression of AP-2beta in different brain regions does not inversely correlate with that of P21728 . These observations taken together indicate that AP-2beta is a repressive transcription factor that acts on the D1AS1 silencer of the P21728 gene via some cell-specific mechanism(s) in Neuro2a . Buspirone anti-dyskinetic effect is correlated with temporal normalization of dysregulated striatal P21728 signalling in DB01235 -treated rats . Dopamine replacement with l-DOPA is the most effective therapy in Parkinson 's disease . However , with chronic treatment , half of the patients develop an abnormal motor response including dyskinesias . The specific molecular mechanisms underlying dyskinesias are not fully understood . In this study , we used a well-characterized animal model to first establish the molecular differences between rats that did and did not develop dyskinesias . We then investigated the molecular substrates implicated in the anti-dyskinetic effect of buspirone , a 5HT1A partial agonist . Striatal protein expression profile of dyskinetic animals revealed increased levels of the dopamine receptor (DR)D3 , ΔFosB and phospho (p)CREB , as well as an over-activation of the P21728 signalling pathway , reflected by elevated ratios of phosphorylated Q9UD71 and P28482 . Buspirone reduced the abnormal involuntary motor response in dyskinetic rats in a dose-dependent fashion . Buspirone ( 4 mg/kg ) dramatically reduced the presence and severity of dyskinesias ( by 83 % ) and normalized Q9UD71 and P28482 phosphorylation ratios , while the increases in P35462 , ΔFosB and pCREB observed in dyskinetic rats were not modified . Pharmacological experiments combining buspirone with 5HT1A and P35462 antagonists confirmed that normalization of both pDARPP32 and pERK2 is required , but not sufficient , for blocking dyskinesias . The correlation between pDARPP32 ratio and dyskinesias was significant but not strong , pointing to the involvement of convergent factors and signalling pathways . Our results suggest that in dyskinetic rats P35462 striatal over-expression could be instrumental in the activation of P21728 -downstream signalling and demonstrate that the anti-dyskinetic effect of buspirone in this model is correlated with P21728 pathway normalization . Differential treatment regimen-related effects of cannabinoids on D1 and D2 receptors in adolescent and adult rat brain . Animal studies suggest differential effects of cannabinoids on dopamine-related behaviours in adolescence and adulthood however few studies have investigated the underlying neurochemical effects of cannabinoids during adolescence . The aim of the present study was to compare the effects of treatment with the synthetic cannabinoid , HU210 , on dopamine receptor density in adolescent and adult rats . Adolescent ( postnatal day ( P01160 ) 35 ) and adult ( P01160 70 ) rats received a single dose of 100μg/kg HU210 or 25 , 50 or 100μg/kg HU210 for 4 or 14 days . P21728 ( D1R ) or D2 receptor ( D2R ) density was measured in the medial and lateral ( CPUL ) caudate putamen , nucleus accumbens , olfactory tubercle ( TU ) and substantia nigra ( D1R only ) using in vitro autoradiography . D1R and D2R densities were 1.6-1.7- and 1.1-1.4-fold higher respectively in adolescent control rats compared to adults . In adult rats , D1R density was increased by 1.2- and 1.3-fold ( p < 0.05 ) in CPUL and TU respectively compared to controls , after 14 days of HU210 treatment . A significant overall effect of treatment ( p < 0.05 ) on D2R density was also observed in adults after the single dose and 4 and 14 days administration of HU210 . In adolescents , an overall effect of treatment on D1R density after a single exposure to HU210 was seen ( p=0.0026 ) but no changes in D1R or D2R densities were observed in other treatment groups . These results suggest that the adolescent rat brain does not display the same compensatory mechanisms activated in the adult brain following cannabinoid treatment . The interleukin 10 promoter haplotype ACA and the long-form variant of the P21917 uVNTR polymorphism are associated with vulnerability to schizophrenia . A total of 934 patients with schizophrenia and 433 controls were genotyped for the interleukin-10 ( P22301 ) promoter and P21917 uVNTR polymorphisms . P21917 long-form variants ( namely , those with ≥5 repeats ) , homozygosity for the 4-repeat allele , and the P22301 haplotype ACA were associated with schizophrenia , respectively . No obvious interactions among the potential polymorphisms were found , which suggests that P22301 and P21917 confer vulnerability to schizophrenia independently . Effects of chronic ' Binge ' cocaine administration on plasma DB01285 and corticosterone levels in mice deficient in Q9UD71 . The product of the Q9UD71 gene mediates intracellular signals initiated by the binding of dopamine to its receptors . Cocaine administration leads to increased activation of dopamine receptors , and causes activation of the stress-responsive hypothalamic-pituitary-adrenal ( Q9Y251 ) axis . We determined the effects of chronic ' binge ' pattern cocaine on Q9Y251 activity in mice containing a targeted disruption of the Q9UD71 gene . Mice received three daily injections of cocaine ( 15 mg/kg/injection ) for 14 days , and were sacrificed 30 min after the last injection . We measured the levels of plasma adrenocorticotropin ( DB01285 ) and corticosterone which reflect Q9Y251 activity . In wild-type controls , ' binge ' cocaine administration significantly increased plasma DB01285 and corticosterone levels . In contrast , Q9UD71 -deficient mice failed to show a significant elevation of either plasma DB01285 or corticosterone levels following ' binge ' cocaine . The results indicate that Q9UD71 plays a role in mediating the stimulatory effects of cocaine on the Q9Y251 axis . Nongenomic , glucocorticoid receptor-mediated regulation of serotonin transporter cell surface expression in embryonic stem cell derived serotonergic neurons . Depressive disorders have been linked to the combined dysregulation of the hypothalamus-pituitary-adrenal ( Q9Y251 ) -axis and the serotonergic system . The Q9Y251 -axis and serotonergic ( 5-HT ) neurons exert reciprocal regulatory actions . It has been reported that glucocorticoid-glucocorticoid receptor ( GR ) signaling influences serotonin transporter ( 5-HTT ) transcription but data also points to the fact that 5-HTT expression is regulated nongenomically via redistribution of 5-HTT from the cell surface into intracellular compartments . In order to analyze the acute effects of glucocorticoids on 5-HTT cell surface localization we differentiated serotonergic neurons from mouse embryonic stem ( ES ) cells derived from the C57BL/6N blastocysts . These postmitotic 5-HT neurons express all relevant serotonergic markers following the application of a growth factor-based differentiation protocol . Increasing concentrations of the GR agonist dexamethasone ( DB00514 ) resulted in enhanced , dose-dependent 5-HTT cell surface localization in the presence of the protein synthesis inhibitor cycloheximide already 1h after incubation . Inhibition of GR function by the specific GR-antagonist mifepristone abolished the increase in 5-HTT cell surface localization . Hence , our data account for a nongenomic upregulation of 5-HTT cell surface expression by glucocorticoid-GR interaction which likely constitutes a rapid physiological response to increased levels of glucocorticoids as seen during stress . Taken together , we provide a cellular model to analyze and dissect glucocorticoid- P31645 interactions on a molecular level that corresponds to in vivo animal models using C57BL/6N mice . Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 ) and the retinoid receptors RARs ( P10276 , P10826 and P13631 ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 and the different isoforms of retinoid receptors revealed that P11473 seems to select mainly P10276 to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 . Identification and characterization of mouse Erbb2 gene in silico . The Q9UD71 - Q14849 - O15273 -PNMT-MGC9753- P04626 -MGC14832- Q14451 locus on human chromosome 17q12 is frequently amplified in human gastric and breast cancer . We have recently identified and characterized human MGC9753 ( also known as wild-type CAB2 ) and mouse Mgc9753 . Here , we identified and characterized mouse Erbb2 gene by using bioinformatics . BLAST programs revealed that mouse AK031099 cDNA was derived from mouse Erbb2 gene . Because AK031099 cDNA showed 806 C --> A nucleotide substitution compared with mouse genome draft sequences and mouse Erbb2 ESTs , the nucleotide sequence of mouse Erbb2 cDNA was determined in silico by correcting 806 A of AK031099 cDNA to C . Nucleotide position 48-3818 of mouse Erbb2 cDNA was the coding region . Mouse Erbb2 gene , consisting of 27 exons , was located within the Ppp1r1b-Grb7 locus on the mouse chromosome 11 . Mouse Erbb2 protein ( 1256 aa ) showed 87.5 % total-amino-acid identity with human P04626 protein , and 95.2 % total-amino-acid identity with rat Erbb2 protein . Mouse Ppp1r1b-Grb7 locus and human Ppp1r1b-Grb7 locus were evolutionarily conserved in the order and the orientation of genes therein . Nucleotide and amino-acid substitution rates of Neurod2 located centromeric to the Ppp1r1b-Grb7 locus were significantly lower than others within the Ppp1r1b-Grb7 locus . This is the first report on the complete coding sequence of mouse Erbb2 gene as well as on the comprehensive comparison of Ppp1r1b-Grb7 locus within the human and mouse genomes . Serial analysis of gene expression predicts structural differences in hippocampus of long attack latency and short attack latency mice . The genetically selected long attack latency ( P38571 ) and short attack latency ( SAL ) mice differ in a wide variety of behavioural traits and display differences in the serotonergic system and the hypothalamus-pituitary-adrenocortical ( Q9Y251 ) -axis . Serial analysis of gene expression ( Q9NXZ1 ) was used to generate a hippocampal expression profile of almost 30 000 genes in P38571 and SAL mice . Using Q9NXZ1 , we found differential expression of 191 genes . Among these were genes involved in growth , signal transduction , and cell metabolism . The Q9NXZ1 study was supported by GeneChip analysis ( Affymetrix ) . Strikingly , both Q9NXZ1 and GeneChips showed a higher expression of numerous cytoskeleton genes , such as cofilin and several tubulin isotypes in P38571 mice . P38571 mice also showed a higher expression of several calmodulin-related genes and genes encoding components of a MAPK cascade , namely raf-related oncogene and P28482 . The findings were confirmed by in situ hybridization . Our results of differential expression of cytoskeleton and signal transduction genes therefore suggest differential regulation of the raf/ P29323 pathway that may be related to structural differences in the hippocampus of P38571 and SAL mice . As stress-related disorders , such as depression , are also linked to differential regulation of the Q9Y251 -axis and the serotonergic system and are associated with altered hippocampal morphology , differential regulation of these genes may be involved in the pathogenesis of these diseases . Expression and prognostic relevance of activated extracellular-regulated kinases ( P27361 /2 ) in breast cancer . Extracellular-regulated kinases ( P27361 , P28482 ) play important roles in the malignant behaviour of breast cancer cells in vitro . In our present study , 148 clinical breast cancer samples ( 120 cases with follow-up data ) were studied for the expression of P27361 , P28482 and their phosphorylated forms p- P27361 and p- P28482 by immunoblotting , and p- P27361 /2 expression in corresponding paraffin sections was analysed by immunohistochemistry . The results were correlated with established clinical and histological prognostic parameters , follow-up data and expression of seven cell-cycle regulatory proteins as well as P03956 , P14780 , P05121 and AP-1 transcription factors , which had been analysed before . High p- P27361 expression as determined by immunoblots correlated significantly with a low frequency of recurrences and infrequent fatal outcome ( P = 0.007 and 0.008 ) and was an independent indicator of long relapse-free and overall survival in multivariate analysis . By immunohistochemistry , strong p- P29323 staining in tumour cells was associated with early stages ( P = 0.020 ) , negative nodal status ( P = 0.003 ) and long recurrence-free survival ( P = 0.017 ) . In contrast , expression of the unphosphorylated kinases P27361 and P28482 was not associated with clinical and histological prognostic parameters , except a positive correlation with oestrogen receptor status . Comparison with the expression of formerly analysed cell-cycle- and invasion-associated proteins corroborates our conclusion that activation of P27361 and P28482 is not associated with enhanced proliferation and invasion of mammary carcinomas . DB01259 -mediated cyclooxygenase-2 expression via epidermal growth factor receptor/ Q15717 interaction enhances the aggressiveness of triple-negative breast cancer cells . DB01259 , a dual epidermal growth factor receptor ( P00533 ) /human epidermal growth factor receptor 2 ( P04626 ) kinase inhibitor , showed clinical benefits in advanced P04626 -positive breast cancer patients . Because some triple-negative breast cancers ( TNBCs ) frequently overexpress P00533 , the antitumor activity of lapatinib in such diseases was also tested . However , the results showed a worse event-free survival rate . It remains unknown whether and how lapatinib elicits the aggressiveness of such cancer cells . In this study , our results demonstrated that lapatinib facilitated axillary and lung metastases of triple-negative MDA-MB-231 breast cancer cells without affecting their viability , leading to worse survival in orthotopic xenograft mice . The lapatinib-increased motility was attributed by the elevation of P00533 through the downregulation of microRNA-7 and by the subsequent overexpression of cyclooxygenase-2 ( P35354 ) . Strikingly , independent of its kinase activity , the elevated P00533 at least partly stabilized P35354 expression by enhancing the binding of Q15717 to P35354 mRNA . Our results suggest that lapatinib may increase the migration and invasion of MDA-MB-231 cells by upregulating P00533 and P35354 through the downregulation of microRNA-7 , providing a potential explanation for the worse clinical outcome of TNBC patients who receive lapatinib-based treatment . These findings also shed new light on the molecular mechanism of P35354 mRNA stabilization by P00533 in a kinase-independent manner . Targeted deletion of P28482 in cardiomyocytes attenuates hypertrophic response but provokes pathological stress induced cardiac dysfunction . Mitogen-activated protein kinases ( MAPKs ) are involved in the regulation of cardiac hypertrophy and myocyte survival . Extracellular signal regulated protein kinase 1 and 2 ( P27361 /2 ) are key components in the MAPK signaling pathways . Dysfunction of P27361 /2 in congenital heart diseases ( Noonan syndrome and LEOPARD syndrome ) leads to cardiac hypertrophy . P28482 contributes 70 % of protein content to total P27361 /2 content in myocardium ; however , the specific role of P28482 in regulating cardiac hypertrophy is yet to be further defined . To investigate the specific role of P28482 played in the cardiomyocytes , we generated and examined mice with cardiomyocyte-specific deletion of the erk2 gene ( P28482 (cko) mice ) . Following short-term pathological hypertrophic stresses , the mutant mice showed attenuated hypertrophic remodeling characterized by a blunted increase in the cross-sectional area of individual myocytes , downregulation of hypertrophic foetal gene markers ( P01160 and DB04899 ) , and less interstitial fibrosis . However , increased cardiomyocyte apoptosis was observed . Upon prolonged stimulation , P28482 (cko) mice developed deterioration in cardiac function . However , absence of P28482 did not affect physiological hypertrophy induced by 4weeks of swimming exercise . These results revealed an essential role for P28482 in cardiomyocytes in the development of pathological hypertrophic remodeling and resistance to cell death . P12821 inhibitors could be therapeutic for antisocial personality disorder . Antisocial personality traits are an important topic for research . The societal cost of these behaviors encourages efforts at a better understanding of central nervous system causes . Catecholamine genes are being studied to facilitate this understanding , and some tentative findings are being reached about several of these genes . It seems that many genes play a role to produce antisocial behaviors so complexity of elucidating each gene is obvious . One conclusion that could be drawn from the current research findings is that DA2 like receptors ( P14416 , P35462 , P21917 ) with alleles that decrease neurotransmission are facilitatory of antisocial behaviors . DA2 like receptors cause neuronal firing to inhibit many peripheral functions through adenylyl cyclase inhibition . When these receptors are less active by genetically decreased density , lower affinity , or by low dopamine levels as final common pathways then inhibition is released and a state of disinhibition can be said to describe this state . Peripheral metabolism is increased and behavioral activation is noted . P00797 is disinhibited in this setting thus allowing sympathetic nervous system activation . The fight or flight behaviors thus produced , in the extreme , would be the setting of antisocial behavior . Research validates this hypothesis . Understanding this final common pathway toward antisocial behavior should lead to better treatment for individuals with this pattern of behavior before they have caused harm to themselves and others . P12821 inhibitors are well tolerated drugs used in the treatment of hypertension and heart failure and would also treat antisocial behavior disorders . P35462 stimulation underlies the development of DB01235 -induced dyskinesia in animal models of Parkinson 's disease . Development of DB01235 -induced dyskinesia ( LID ) remains a major problem in the long-term treatment of Parkinson 's disease ( PD ) . Sensitization to DB01235 correlates with ectopic expression of D3 dopamine receptors in the striatum , implicating D3 receptors in development of LID . We demonstrate that the selective D3 antagonist S33084 abolishes development of LID over 30 days in MPTP-lesioned marmosets without effecting the anti-parkinsonian actions of DB01235 . Furthermore , following a 14 day washout , when challenged with DB01235 in the absence of S33084 , these animals continued to exhibit reduced LID . In the 6-OHDA-lesioned rat , S33084 similarly attenuated development of behavioural sensitization to DB01235 . Additionally , DB01235 -induced elevations in striatal pre-proenkephalin-A ( PPE-A ) ( but not PPE-B , phospho[ DB00156 (34)] Q9UD71 , D1 , and D2 receptor mRNA or D3 receptor levels ) were reduced in S33084 treated animals . Our data suggest a role for D3 receptors in the development of LID and suggest that initiating DB01235 treatment with a D3 antagonist may reduce the development of LID in PD . Multiple endocytic signals in the C-terminal tail of the cystic fibrosis transmembrane conductance regulator . The cystic fibrosis transmembrane conductance regulator ( P13569 ) is a DB02527 -dependent protein kinase ( PKA ) -activated chloride channel that is localized to the plasma membrane and endosomal compartment . Endosomal targeting of P13569 is attributed to the DB00135 (1424)-based internalization signal , identified in the C-terminal tail of the channel . Mutation of the DB00135 (1424) residue could partly inhibit the endocytosis of P13569 and its association with the adapter protein P05549 . To reveal additional endosomal targeting signals , site-directed mutagenesis of both a chimaera , composed of a truncated form of interleukin 2 receptor alpha chain ( TacT ) and the C-terminal tail of P13569 ( Ct ) , and the full-length P13569 was performed . Morphological and functional assays revealed the presence of multiple internalization motifs at the C-terminus , consisting of a phenylalanine-based motif ( DB00120 (1413) ) and a bipartite endocytic signal , comprising a tyrosine ( DB00135 (1424) ) and a di- DB00149 -based ( DB00149 (1430)- DB00149 ) motif . Whereas the replacement of any one of the three internalization motifs with alanine prevented the endocytosis of the TacT-Ct chimaera , mutagenesis of DB00120 (1413)- DB00149 impaired the biosynthetic processing of P13569 , indicating that DB00120 (1413) is indispensable for the native structure of P13569 . In contrast , replacement of DB00149 (1430)- DB00149 - and DB00135 (1424)-based signals with alanine increased the cell-surface density of both the chimaeras and P13569 in an additive manner . These results suggest that the internalization of P13569 is regulated by multiple endocytic sorting signals . Dynamic simulations of pathways downstream of P00533 -family , including mutations and treatments : concordance with experimental results . The pathways downstream of ErbB-family proteins are very important in BC , especially when considering treatment with onco-protein inhibitors . We studied and implemented dynamic simulations of four downstream pathways and described the fragment of the signaling network we evaluated as a Molecular Interaction Map . Our simulations , enacted using Ordinary Differential Equations , involved 242 modified species and complexes , 279 reversible reactions and 111 catalytic reactions . Mutations within a single pathway tended to be mutually exclusive ; only inhibitors acting at , or downstream ( not upstream ) , of a given mutation were active . A double alteration along two distinct pathways required the inhibition of both pathways . We started an analysis of sensitivity/robustness of our network , and we systematically introduced several individual fluctuations of total concentrations of independent molecular species . Only very few cases showed significant sensitivity . We transduced the ErbB2 over-expressing BC line , BT474 , with the P01112 ( V12 ) mutant , then treated it with ErbB-family and phosphorylated MEK ( MEKPP ) inhibitors , DB01259 and U0126 , respectively . Experimental and simulation results were highly concordant , showing statistical significance for both pathways and for two respective endpoints , i.e. phosphorylated active forms of P29323 and Akt , p one tailed = .0072 and = .0022 , respectively . Working with a complex 39 basic species signaling network region , this technology facilitates both comprehension and effective , efficient and accurate modeling and data interpretation . Dynamic network simulations we performed proved to be both practical and valuable for a posteriori comprehension of biological networks and signaling , thereby greatly facilitating handling , and thus complete exploitation , of biological data .

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Determination of ancestral allele for possible human cancer-associated polymorphisms . To determine ancestral allele in possible cancer-associated polymorphisms , DNA samples from 10 chimpanzees ( Pan troglodytes ) were sequenced for alleles corresponding to 17 polymorphisms : 8 short tandem repeats [ P18510 ( alias IL-1RA ) variable number tandem repeat ( VNTR ) ; P04818 ( previously TS ) VNTR ; AR CAG repeat ; dinucleotide repeats of P22309 , IGF1 , P01579 ( alias P01579 ) , P03372 ( alias P03372 ) , and P00533 ] and 9 single nucleotide polymorphisms ( P03956 -1607 1G/2G , P08254 -1171 5A/6A , O15527 Ser326Cys , P05091 Gly487Lys , P04637 Arg72Pro , Q9UNQ0 Gln141Lys , P16455 Leu84Phe , P04179 Ala-9Val , and P42898 Ala222Val ) . No chimpanzee polymorphism corresponded to human P18510 VNTR ; the ancestral allele was a repeat lost in humans . Dinucleotide repeat polymorphisms of IGF1 , P01579 , P03372 , and P00533 were shared by chimpanzees , but the length of repeats tended to be longer in humans than in chimpanzees . This tendency was particularly evident for IGF1 . All of the SNPs tested are human-specific nucleotide changes . The ancestral allele 7A was shown to be lost in P08254 -1171 5A/6A . Thus , all of the possible cancer-associated polymorphisms tested have human-specific alleles , and the ancestral allele is lost in three polymorphisms ( P18510 VNTR , P22309 CA repeat , and P08254 -1171 5A/6A ) , suggesting a possible involvement of human-specific alleles in cancer susceptibility . Contribution of cubilin and amnionless to processing and membrane targeting of cubilin-amnionless complex . O60494 is a peripheral apical membrane receptor for multiple ligands that are taken up in several absorptive epithelia . Recently , amnionless ( Q9BXJ7 ) was identified to form a functional receptor complex with cubilin . By expression in transfected polarized MDCK cells of Q9BXJ7 and several cubilin fragments , including a functional " mini " version of cubilin , the processing , sorting , and membrane anchoring of the complex to the apical membrane were investigated . The results show that truncation mutants , including the N-terminal domain of cubilin , did not appear at the plasma membrane but instead were retained in the endoplasmic reticulum or partially secreted into the medium . Coexpression with Q9BXJ7 led to efficient transport to the apical cell surface of the cubilin constructs , which included the P01133 domains , and prevented release into the medium . Q9BXJ7 co-precipitated with cubilin and co-localized with cubilin at the apical cell surface . Apical sorting was observed for a broad set of nonoverlapping cubilin fragments without the N-terminal region , in the absence of Q9BXJ7 . The preference for apical sorting disappeared when glycosylation was inhibited by tunicamycin . In conclusion , it is shown that both units contribute to the processing of the cubilin- Q9BXJ7 complex to the apical membrane : Q9BXJ7 interacts with the P01133 domains of cubilin and is responsible for membrane attachment and export of the complex from the endoplasmic reticulum , whereas the extracellular cubilin molecule is responsible for apical sorting of the complex in a carbohydrate-dependent manner . Suppression of proliferation and migration in highly-metastatic lung cancer cells as well as tumor growth by a new synthesized compound TBrC and its molecular mechanisms of action . To develop new anticancer agents has been considered as a useful and necessary strategy to suppress highly-metastatic lung cancer , the leading cause of cancer-related deaths in the world . In this study , we synthesized a new compound ethyl 6-bromocoumarin-3-carboxylyl L-theanine ( TBrC ) and studied the anticancer activity of TBrC and its molecular mechanisms of action . Our results show that TBrC remarkably inhibits the proliferation and migration in highly-metastatic lung cancer cells by inducing apoptosis and cell cycle arrest as well as regulating related protein expressions . Further study indicated that TBrC not only enhances the protein levels of Bax , cytosolic cytochrome c , caspase-3 and P09874 but also reduces the protein expressions of Bcl-2 , cyclin D1 , P17948 and NF-κB as well as inhibits the phosphorylation and expressions of P35968 and Akt in the cancer cells . More importantly , TBrC displays strong suppression of highly-metastatic tumor growth and reduces the tumor weight by 61.6 % in tumor-bearing mice without toxicity to the mice . Our results suggest that TBrC suppresses the proliferation and migration of lung cancer cells via VEGFR-Akt-NF-κB signaling pathways ; TBrC may have a wide therapeutic and/or adjuvant therapeutic application in the treatment of lung cancer . The preclinical development of regorafenib for the treatment of colorectal cancer . INTRODUCTION : The DB01367 -RAF-MEK- P29323 pathway is one of the best characterized kinase cascades . During the exploration of small molecules that inhibit P04049 kinase , regorafenib ( DB08896 ) was discovered as a multikinase inhibitor which demonstrated anti-cancer , anti-angiogenic , and apoptotic activities in metastatic colorectal cancer . This was not the first multikinase inhibitor discovered for the disease ; indeed , before regorafenib was approved by FDA as a multikinase inhibitor for metastatic colorectal cancer in 2012 , sorafenib ( BAY 43-9006 ) had already been developed to be the first in the world as a multikinase inhibitor for malignancy . Indeed , the only difference between the two compounds is fluorine bound to its proximal phenyl ring although the end result is a considerably different profile , both as a kinase inhibitor as well as in its clinical application . AREAS COVERED : In this drug discovery case history , the authors review the design , discovery , and development of both regorafenib and sorafenib from back in the 1990s . Furthermore , the authors highlight the drug 's anti-cancer and anti-angiogenic properties as well as its efficacy , safety pharmacology and toxicology based on FDA documents . EXPERT OPINION : In order to better predict the efficacy of kinase inhibitors and to utilize them more efficiently , our understanding of drug discovery , the approaches for kinase profiling , and technologies needed for their development are paramount . Indeed , the authors believe that the field should better explore the use of predictive biomarkers that might be able to better assess these therapeutics . Pharmaceutical scientists must also consider the cost effectiveness of the targeted agents developed as a number of the drugs developed are very expensive . Gambogic acid inhibits angiogenesis through suppressing vascular endothelial growth factor-induced tyrosine phosphorylation of P35968 /Flk-1 . Previous studies revealed that gambogic acid ( GA ) , the major active ingredient of gamboge , a brownish to orange resin exuded from Garcinia hanburryi tree in Southeast Asia , possessed significant anticancer activity both in vitro and in vivo . In this study , we explored the high antiangiogenic activities of GA for the first time . GA inhibits the P15692 -stimulated proliferation , migration and tube formation of human umbilical vein endothelial cells ( HUVECs ) as well as microvessel sprouting from rat aortic rings in vitro . Moreover , GA inhibits vessel growth in matrigel plugs and P62158 in vivo and transplanted tumor in mice . The results also indicated that GA decreases P15692 production of cultured tumor cells and inhibits P15692 -induced tyrosine phosphorylation of P35968 /Flk-1 . This inhibition of receptor phosphorylation is correlated with a significant decrease in P15692 -triggered phosphorylated forms of P29323 , AKT and p38 . Taken together , these findings strongly suggest that GA might be a structurally novel angiogenesis inhibitor . Activity of nilotinib ( Q9BXJ7 -107 ) alone in advanced gastrointestinal stromal tumors progressing on imatinib and sunitinib . Case report . DB04868 is a novel tyrosine kinase inhibitor targeting P10721 ( CD117 ) , P09619 and P11274 - P00519 and inhibiting the proliferation of both imatinib-sensitive and imatinib-resistant cells in vitro . DB04868 , alone or in combination with imatinib , has promising activity in imatinib-resistant patients with advanced gastrointestinal stromal tumors ( GISTs ) , including those who progressed on sunitinib and other tyrosine kinase inhibitors . We describe the beneficial effect of nilotinib 400 mg b.i.d. orally in a 53-year-old patient with metastatic GISTs who had radiologically confirmed disease progression on both imatinib ( 800 mg/day ) and sunitinib ( 50 mg/day ) . A positron emission tomography-computerized tomography evaluation showed marked decreases in (18)F- DB09150 uptake in the liver and mesentery region after 3 months of therapy with nilotinib . DB04868 is an option for patients with advanced GISTs progressing on both imatinib and sunitinib . The effects of five alkaloids from Bulbus Fritillariae on the concentration of DB02527 in P29320 cells transfected with muscarinic M(2) receptor plasmid . The aim of this study was to investigate the effects of five alkaloids , namely verticine , verticinone , imperialine , imperialine-3beta-D-glucoside , and puqietinone , purified from Bulbus Fritillariae and used as an antitussive drug in traditional Chinese medicine , on their antimuscarinic M(2) function and the DB02527 level of P29320 cells transfected with muscarinic M(2) receptor plasmid . By transfecting the P29320 cells with the method of calcium phosphate co-precipitation and screening with G418 , the cells stably expressing M(2) receptor were identified . The expression of M(2) receptor in P29320 cells was confirmed by both RT-PCR and western blot . The DB02527 level in the treated cells was analyzed with RIA method ( (125)I- DB02527 P10721 ) . And the results suggested that the five alkaloids could significantly elevate the DB02527 concentration in the P29320 cells transfected with muscarinic M(2) receptor plasmid ( p < 0.01 ) . Functional cross talk between P61073 and P09619 on glioblastoma cells is essential for migration . Glioblastoma ( GBM ) is the most common and aggressive form of brain tumor , characterized by high migratory behavior and infiltration in brain parenchyma which render classic therapeutic approach ineffective . The migratory behaviour of GBM cells could be conditioned by a number of tissue- and glioma-derived cytokines and growth factors . Although the pro-migratory action of P48061 on GBM cells in vitro and in vivo is recognized , the molecular mechanisms involved are not clearly identified . In fact the signaling pathways involved in the pro-migratory action of P48061 may differ in individual glioblastoma and integrate with those resulting from abnormal expression and activation of growth factor receptors . In this study we investigated whether some of the receptor tyrosine kinases commonly expressed in GBM cells could cooperate with P48061 / P61073 in their migratory behavior . Our results show a functional cross-talk between P61073 and P09619 which appears to be essential for GBM chemotaxis . DB08896 for gastrointestinal malignancies : from preclinical data to clinical results of a novel multi-target inhibitor . Intracellular signals for cancer cell growth , proliferation , migration , and survival are frequently triggered by protein tyrosine kinases ( TKs ) . The possibility of disrupting core disease pathways has led to development and widespread clinical use of specific TK inhibitors that in the past decade have markedly changed treatment strategies and impacted on overall outcomes . However , intrinsic resistance may limit the benefit of these drugs , and multiple escape routes compensate for the inhibited signaling . The disruption of several points of the same pathway and the simultaneous interference with different intracellular oncogenic processes have both been recognized as valuable strategies to maximize the therapeutic potential of this class of agents . In this scenario , regorafenib has emerged as a novel , orally active , multitarget compound with potent activity against a number of angiogenic and stromal TKs , including vascular endothelial growth factor receptor 2 ( P35968 ) , tyrosine kinase with immunoglobulin-like and P01133 -like domains 2 ( P35590 -2 ) , fibroblast growth factor receptor 1 ( P11362 ) , and platelet-derived growth factor receptor ( P09619 ) . Moreover , the drug has the capability of blocking P10721 , P07949 and V600 mutant P15056 . Starting from interesting preclinical results , this review describes the clinical development of regorafenib in gastrointestinal malignancies , focusing on data derived from cutting edge clinical trials that have provided evidence of efficacy in pretreated patients with advanced colorectal cancer or gastrointestinal stromal tumors . Casitas B-lineage lymphoma mutants activate AKT to induce transformation in cooperation with class III receptor tyrosine kinases . In addition to overexpression and the occurrence of activating mutations , receptors can be aberrantly activated by impaired downregulation . In this study , we show that an oncogenic mutant of the ubiquitin ligase casitas B-lineage lymphoma ( P22681 ; CBLΔexon8 ) , which is found in acute myeloid leukemia patients , predominantly cooperates with receptor tyrosine kinase ( RTK ) class III receptors ( P16234 , P09619 , P10721 , and P36888 ) , but not with non-class III RTKs or cytokine receptors , to induce P08700 -independent growth of Ba/ P13726 cells . In cells coexpressing RTK class III/CBLΔexon8 , receptor internalization was delayed , and cells were protected from apoptosis after cytokine withdrawal . Ligand-stimulated Ba/ P13726 cells and acute myeloid leukemia cell lines coexpressing the P22681 deletion mutant and P36888 showed enhanced AKT phosphorylation . Combined pharmacologic inhibition of the PI3K/AKT pathway and P36888 had an additive effect on cell proliferation . The transforming potential of the P22681 mutant was completely abolished by the mutation of the P22681 PTB domain and was decreased by the mutation of tyrosines 589 and 591 in the juxtamembrane domain of P36888 . A constitutively active P31749 mutant ( E17K ) recapitulated the phenotype induced by the P22681 deletion mutant in Ba/ P13726 cells . This study reveals P36888 - P22681 interaction sites and the AKT pathway as critical mediators of transformation by oncogenic P22681 mutants . [ New insights into hypereosinophilic syndromes ] . Hypereosinophilic syndrome ( DB09106 ) is characterized by chronic unexplained eosinophilia with organ involvement . The concept of DB09106 as a single disease entity is being challenged by the recent identification of multiple underlying molecular mechanisms . DB09106 can directly affect the eosinophil lineage ( often linked to a fusion gene Q6UN15 - P16234 , and corresponding in this case to chronic eosinophilic leukemia ) , or the lymphoid lineage , where eosinophilia is secondary to expansion of a T cell subset overproducing interleukin-5 , a cytokine involved in eosinophilopoiesis . These recent discoveries have legitimized the use of tyrosine kinase inhibitors such as imatinib , which , by inhibiting P16234 , have transformed the prognosis of chronic eosinophilic leukemia , and also the use of monoclonal anti- P05113 antibodies , which are promising treatment for steroid-dependent DB09106 . [ DB00391 in the management of functional dyspepsia and delayed gastric emptying ] . DB00391 is a sulpiride isomer that exerts its prokinetic action through a dual mechanism : 1 ) as a P14416 antagonist and 2 ) as a serotonin 5HT(4) receptor agonist , conferring this drug with a cholinergic effect . At a dosage of 25mg three times daily , levosulpiride accelerates gastric and gallbladder emptying . Clinical trials have shown that this agent is more effective than placebo in reducing the symptoms of dyspepsia , while comparative studies have demonstrated that its effect is similar or superior to that of other dopamine antagonists . The safety profile of levosulpiride is good and the frequency of adverse events is similar to that of other D(2) dopamine antagonists . Therefore , this drug is a useful therapeutic option in the management of patients with functional dyspepsia , as well as in those with delayed gastric emptying . Variants of dopamine and serotonin candidate genes as predictors of response to risperidone treatment in first-episode schizophrenia . AIMS : Abnormalities in dopaminergic and serotonergic transmission systems are thought to be involved in the pathophysiology of schizophrenia and the mechanisms underlying the therapeutic effects of antipsychotics . We conducted a pharmacogenetic study to evaluate whether variants in dopamine-related genes ( P21728 - P21918 , P31749 and GSK3beta ) and serotonin receptor genes ( P08908 , P28222 , P28221 , P28223 , P28335 , P50406 and P34969 ) can be used to predict the efficacy of risperidone treatment for schizophrenia . MATERIALS & METHODS : A total of 120 first-episode neuroleptic-naive schizophrenia patients were treated with risperidone monotherapy for 8 weeks and clinical symptoms were evaluated by the Positive and Negative Syndrome Scale . RESULTS : Among the 30 variants that we examined , two SNPs in P14416 ( -241A > G [ rs1799978 ] and TaqIA [ rs1800497 ] ) and two SNPs in P31749 ( P31749 -SNP1 [ rs3803300 ] and P31749 -SNP5 [ rs2494732 ] ) were significant predictors of treatment response to risperidone . CONCLUSION : These data suggest that the SNPs in P14416 and P31749 may influence the treatment response to risperidone in schizophrenia patients . Mast cells and eosinophils in mastocytosis , chronic eosinophilic leukemia , and non-clonal disorders . Mast cells and eosinophils often travel in the same biologic circles . In non-clonal states , such as allergic and inflammatory conditions , cell-to-cell contact and the pleiotropic actions of multiple cytokines and chemokines , derived from local tissues or mast cells themselves , foster the co-recruitment of these cells to the same geographic cellular niche . While eosinophils and mast cells serve critical roles as part of the host immune response and in maintenance of normal homeostasis , these cell types can undergo neoplastic transformation due to the development of clonal molecular abnormalities that arise in early hematopoietic progenitors . The dysregulated tyrosine kinases , D816V P10721 and Q6UN15 - P16234 , are the prototypic oncogenic lesions resulting in systemic mastocytosis ( SM ) and chronic eosinophilic leukemia , respectively . We review the pathobiology of these myeloproliferative neoplasms ( MPNs ) with a focus on the relationship between mast cells and eosinophils , and discuss murine models , which further elucidate how the phenotype of these diseases can be influenced by stem cell factor ( P21583 ) and expression of the potent eosinophilopoietic cytokine , interleukin-5 ( P05113 ) . Therapy of SM and Q6UN15 - P16234 -positive disease and the prognostic relevance of increased peripheral blood and tissue mast cells in hematolymphoid malignancies will also be addressed . Modulatory effects of heparin and short-length oligosaccharides of heparin on the metastasis and growth of LMD MDA-MB 231 breast cancer cells in vivo . Expression of the chemokine receptor P61073 allows breast cancer cells to migrate towards specific metastatic target sites which constitutively express P48061 . In this study , we determined whether this interaction could be disrupted using short-chain length heparin oligosaccharides . Radioligand competition binding assays were performed using a range of heparin oligosaccharides to compete with polymeric heparin or heparan sulphate binding to I(125) P48061 . DB01109 dodecasaccharides were found to be the minimal chain length required to efficiently bind P48061 ( 71 % inhibition ; P < 0.001 ) . These oligosaccharides also significantly inhibited P48061 -induced migration of P61073 -expressing LMD MDA-MB 231 breast cancer cells . In addition , heparin dodecasaccharides were found to have less anticoagulant activity than either a smaller quantity of polymeric heparin or a similar amount of the low molecular weight heparin pharmaceutical product , DB06822 . When given subcutaneously in a SCID mouse model of human breast cancer , heparin dodecasaccharides had no effect on the number of lung metastases , but did however inhibit ( P < 0.05 ) tumour growth ( lesion area ) compared to control groups . In contrast , polymeric heparin significantly inhibited both the number ( P < 0.001 ) and area of metastases , suggesting a differing mechanism for the action of polymeric and heparin-derived oligosaccharides in the inhibition of tumour growth and metastases . Phosphorylation of calmodulin by the epidermal-growth-factor-receptor tyrosine kinase . An epidermal-growth-factor( P01133 )-receptor preparation isolated by calmodulin-affinity chromatography from rat liver plasma membranes is able to phosphorylate calmodulin . P62158 phosphorylation was enhanced 3-8-fold by P01133 , was dependent on the presence of a polycation or basic protein and was inhibited by micromolar concentrations of Ca2+ . Phosphate incorporation into calmodulin occurs predominantly on tyrosine residues . Partial proteolysis of phosphocalmodulin by thrombin identifies Tyr99 , located in the third calcium-binding domain of calmodulin , as the phosphorylated residue . Stoichiometric measurements show a 32P/calmodulin molar ratio of approximately 1 when optimal phosphorylation conditions are used . Critical appraisal of the use of regorafenib in the management of colorectal cancer . The lack of valid clinical management options for patients affected by metastatic colorectal cancer , which has progressed after all approved standard treatments , has lead to research into new active molecules . DB08896 is an oral small-molecule multi kinase inhibitor , binding to several intracellular kinases , with powerful inhibitory activity against vascular endothelial growth factor receptors ( P17948 , P35968 , and P35916 ) , platelet-derived growth factor receptor , fibroblast growth factor receptor 1 , Raf , P35590 -2 , and the kinases P10721 , P07949 , and P15056 . The antitumor activity of regorafenib has been tested in vitro and in vivo , and inhibition of tumor growth has been observed in several cancer models , particularly colorectal cancer and gastrointestinal stromal tumors . The most frequent adverse events of grade 3 or higher related to regorafenib were hand-foot skin reaction , fatigue , diarrhea , hypertension , and rash or desquamation . Only a few Phase I-II trials , and most recently a Phase III trial in pretreated colorectal cancer , have been carried out to date . Several ongoing trials are testing the efficacy of regorafenib in combination with chemotherapy . At this point in time , regorafenib is the first small-molecule tyrosine kinase inhibitor to gain approval by the US Food and Drug Administration for pretreated metastatic colorectal cancer patients . Inhibition of proliferation and migration of luminal and claudin-low breast cancer cells by P09619 inhibitors . BACKGROUND : Platelet-derived growth factors ( PDGFs ) bind to two receptors , PDGFRα and PDGFRβ to mediate cell proliferation , migration and survival . Although epithelial cells typically do not express high levels of PDGFRs , their expression has been reported to increase in breast cancer cells that have undergone epithelial to mesenchymal transition . METHODS : P09619 signaling was inhibited using DB01268 malate , Imatinib mesylate or DB08896 in murine and human luminal-like and claudin-low mammary tumor cell lines or Masitinib in only the human cell lines . A scratch wound assay was used to assess tumor cell migration while immunofluorescence for phosphorylated histone H3 or cleaved caspase 3 was used to determine tumor cell proliferation and apoptosis , respectively . RESULTS : DB01268 and DB08896 , but not Imatinib , were capable of significantly inhibiting the migration of both murine and human luminal-like and claudin-low breast cancer cells while Masitinib inhibited migration in both human breast cancer cell lines . DB01268 but not DB08896 or Imatinib also significantly suppressed tumor cell proliferation in all four cell lines tested while Masitinib had no significant effect on human breast cancer cell proliferation . None of the P09619 inhibitors consistently regulated mammary tumor cell apoptosis . CONCLUSION : DB01268 , DB08896 and Masitinib may prove clinically useful in inhibiting breast cancer cell migration and metastasis while only DB01268 ( and possibly DB08896 in some breast cancer subtypes ) is effective at inhibiting both migration and proliferation of breast cancer cells . Lessons learned from the irinotecan metabolic pathway . DB00762 , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN-38 . SN-38 acts as a P11387 poison . DB00762 has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN-38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 is subjected to be shunted between P08684 mediated oxidative metabolism to form two inactive metabolites P25054 or NPC and tissue carboxylesterase mediated hydrolysis to form SN-38 which is eventually detoxified via glucuronidation by P22309 to form SN-38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e.g. , P08684 , P20815 , P22309 ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John 's Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e.g. , Pgp , Q9UNQ0 , MRP1 , Q92887 ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP3A-mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent . Identification of new candidate therapeutic target genes in triple-negative breast cancer . Triple-negative breast cancer ( TNBC ) is a subgroup of breast cancer that is negative for estrogen and progesterone receptor and P04626 protein expression . It is characterized by its aggressive behavior and by the lack of targeted therapies . To identify new therapeutic targets in TNBC , we used real-time quantitative RT-PCR to analyze 63 TNBC samples in terms of their mRNA expression of 26 genes coding for the major proteins currently targeted by drugs used to treat other cancers or undergoing clinical trials in breast cancer . Six of the 26 genes tested ( P15692 , P12931 , P09874 , Q05397 , P04049 , and P22607 ) were significantly upregulated in 13 % to 46 % of the TNBCs . None of the 6 genes was specifically upregulated in the TNBCs compared with 3 other classical breast tumor subtypes . No association was observed between overexpression of these 6 genes ( except for P22607 ) and P42336 mutation status . These results confirm the interest of targeting P15692 and P09874 in ongoing clinical trials in TNBC patients and also identify new target genes ( P12931 , Q05397 , P04049 , and P22607 ) . Clinical trials could be initiated easily with existing drugs . Our results also suggest that these target genes might serve as predictive biomarkers of the TNBC treatment response . Cord blood stem-cell-derived dendritic cells generate potent antigen-specific immune responses and anti-tumour effects . The aim of the present study was to investigate whether CBSCs [ ( umbilical ) cord blood stem cells ] can be a new source of DCs ( dendritic cells ) , which can generate more potent antigen-specific immune responses and anti-tumour effects . CBSCs and PBMCs ( peripheral blood mononuclear cells ) were collected , cultured and differentiated into DCs . Surface markers , secreting cytokines , antigen-presentation activity , antigen-specific cell-mediated immunity and cytotoxic killing effects induced by these two DC origins were evaluated and compared . CBSCs were expanded ~17-fold by ex vivo culture . The expression of surface markers in CBSC-derived DCs were higher than those in PBMC-derived DCs treated with LPS ( lipopolysaccharide ) . The CBSC-derived DCs mainly secreted IL (interleukin)-6 , P22301 and P01375 ( tumour necrosis factor ) -α , whereas PBMC-derived DCs mainly secreted P05113 and IFN (interferon)-γ . The CBSC-derived DCs had better antigen-presentation abilities when stimulated with LPS or P01375 -α , induced higher numbers of IFN-γ-secreting antigen-specific CD8+ T-cells , as assessed using an ELISpot ( enzyme-linked immunosorbent spot ) assay , and stimulated more potent antigen-specific CTL ( cytotoxic T-cell ) activities ( P < 0.01 , one-way Q9UNW9 ) . CBSC-derived DCs had quicker and greater P29323 ( extracellular-signal-regulated kinase ) and Akt phosphorylation , and weaker p38 phosphorylation , than PBMC-derived DCs when stimulated with LPS . In conclusion , CBSC-derived DCs have the ability to induce stronger antigen-specific immunity and more potent anti-tumour effects and therefore could be a good source of DCs for use in DC-based cancer vaccines and immunotherapy . The interleukin-5/receptor interaction activates Lyn and Jak2 tyrosine kinases and propagates signals via the Ras- P04049 - Q96HU1 kinase and the Jak- P35610 pathways in eosinophils . We have shown that the interaction of interleukin ( IL ) -5 with the receptor activates Lyn tyrosine kinase within 1 min and Jak2 tyrosine kinase within 1-3 min . P05113 also stimulates GTP binding to P01112 . The signal is subsequently propagated through the activation of P04049 , MEK , and Q96HU1 kinases as shown by their increased autophosphorylation in vitro and phosphorylation in situ . Jak2 kinase has been shown to phosphorylate P35610 nuclear proteins . The activation of P35610 nuclear factors was studied by electrophoretic mobility shift assay using a gamma activation site ( GAS ) probe . We found that P05113 induces two GAS-binding proteins in eosinophils , one of which is P42224 . We conclude that P05113 induced signals are propagated through two distinct pathways : ( 1 ) Lyn --> Ras --> P04049 --> MEK --> Q96HU1 kinase and ( 2 ) Jak2 --> P42224 . [ Genetic and epigenetic changes in colorectal cancer and genetic testing for personalized medicine ] . Recent studies have uncovered molecular pathways of colorectal cancer , including the chromosomal instability pathway and microsatellite pathway . In addition , according to genetic and epigenetic profiles , colorectal cancer can be subclassified into 3 distinct groups , named the CpG island methylator phenotype ( CIMP ) 1 , CIMP2 , and CIMP negative . CIMP1 is characterized by MSI and P15056 mutations and rare P01116 and p53 mutations . CIMP2 is associated with P01116 mutations and rare MSI , P15056 , or p53 mutations . CIMP negative cases have a high rate of p53 mutations and lower rates of MSI or mutations of P15056 or P01116 . Regarding genetic testing for personalized medicine for colorectal cancer , uridine disphosphate glucuronosyl transferase 1(UGT1) and P01116 tests are available . DB00762 is one of the most effective chemotherapeutic agents in the treatment of metastatic colorectal cancer . The prodrug irinotecan is biotransformed by carboxylesterase into its active metabolite SN-38 , which is inactivated by UGT1 into the inactive compound SN-38G . Here we discuss P22309 gene polymorphism as a predictor of toxicity . The epidermal growth factor ( P00533 ) plays an important role in the development and progression of colorectal cancer . P01116 serves as a mediator between extracellular ligand binding and intracellular transduction of signals from P00533 to the nucleus . Activating P01116 mutations has been identified as a predictor of resistance to P00533 -directed antibodies such as cetuximab . Here we discuss the current understanding of P01116 mutations and the therapeutic effect of cetuximab . P09874 deletion promotes subventricular zone neural stem cells toward a glial fate . Identification of critical factors involved in oligodendroglial fate specification from endogenous neural stem cells is relevant to the development of therapeutic interventions that aim to promote remyelination . Here we report a novel role of the DNA repair protein poly-ADP-ribose polymerase-1 ( P09874 ) in regulating the neural stem cell profile in the postnatal mouse forebrain subventricular zone ( SVZ ) . We observed increased expression of Sox2 and Sox10 in the SVZ of postnatal day 11 ( P11 ) P09874 knockout mice . This increase corresponded to increased Olig2 expression in Sox2-positive cells of the P09874 knockout mouse SVZ and decreased Map2abc expression compared with Sox2/Olig2 and Sox2/Map2abc expression in wild-type mice . We noted enhanced expression of proliferating oligodendrocyte progenitor cells ( OPCs ) at the expense of proliferating neuroblasts in the SVZ of P09874 knockout mice , by using Olig1/Ki67/ O43602 , Q99942 /Ki67/ O43602 , and P09619 /BrdU/TuJ1 immunofluorescence labeling . In addition , the percentage of BrdU/Olig2 double-labeled cells increased in the SVZ and corpus callosum of P09874 knockout mice compared with wild-type mice . We also observed a decrease in O43602 -positive cells without a decrease in the overall SVZ area in P09874 knockout mice , further indicating a switch from neuroblast to OPC fate . P09874 knockout mice displayed thinning of MBP expression in the corpus callosum and external capsule , suggesting that the enhanced OPC proliferation in the SVZ might compensate for deficiency in myelination . Together , our results show that P09874 deletion promotes SVZ neural stem cells toward a glial fate and suggest that future studies target P09874 as a potential therapeutic strategy for demyelinating diseases . Peptide-based targeting of the platelet-derived growth factor receptor beta . PURPOSE : The aim of this work is to identify new ligands targeting the platelet-derived growth factor receptor beta ( PDGFRβ ) . PROCEDURES : Biopanning was carried out with a 12-amino-acid phage display library against the recombinant extracellular domain of PDGFRβ . The identified peptide P09619 -P1 was chemically synthesized and labeled with (125)I or (131)I . In vitro studies were performed on the PDGFRβ-expressing cell lines BxPC3 and MCF7 and on PDGFRβ-transfected P29320 cells in comparison to negative control wtHEK293 and CaIX-transfected P29320 cells . Biodistribution experiments were performed in Balb/c nude mice , carrying subcutaneously BxPC3 tumors . RESULTS : In vitro studies demonstrated a higher binding to BxPC3 , MCF7 , and PDGFRβ-tr- P29320 cells in comparison to negative control cell lines . Binding was inhibited up to 90 % by the unlabeled P09619 -P1 peptide . Organ distribution studies revealed a higher accumulation in BxPC3 tumors than in most organs . CONCLUSIONS : P09619 -P1 is a promising candidate for targeting human PDGFRβ . DB08896 ( DB08896 ) : stromal and oncogenic multikinase inhibitor with potential activity in renal cell carcinoma . The introduction of targeted therapies , specifically those that target the P15692 receptor ( VEGFR ) , PDGF receptor ( P09619 ) and the P42345 pathways , has significantly changed the approach to patients with unresectable renal cell cancer ( RCC ) . However , drug resistance develops through bypassing of targeted pathways . DB08896 ( DB08896 ) is a novel bi-aryl urea compound that has potential anti-tumour activity in RCC , as along with targeting P15692 and PDGF receptors , it targets additional kinases associated with alternative pathways of angiogenesis and resistance to P15692 -targeted drugs . Based on a phase II clinical trial , the efficacy outcome of regorafenib in the first-line setting of unresectable RCC appears comparable that of other targeted first-line drugs . However , testing regorafenib in standard phase III trials seems inappropriate in view of its toxic effects . Further assessment of regorafenib should exploit the drug 's ability to inhibit mechanisms of escape from anti-angiogenic treatment through biomarker-driven clinical trials . Isolation of mouse DB05914 on the basis of expression of Sca-1 and P09619 -α . Platelet-derived growth factor receptor α ( P09619 -α ) and stem cell antigen 1 ( Sca-1 ) have recently been identified as selective markers of mouse DB05914 ( MSCs ) . P09619 -α(+)Sca-1(+) ( PαS ) MSCs have augmented growth potential and robust tri-lineage differentiation compared with standard culture-selected MSCs . In addition , the selective isolation of PαS MSCs avoids cellular contamination that can complicate other methods . Here we describe in detail our protocol to isolate PαS MSCs using flow cytometry . In brief , the tibia and femora are isolated and crushed using a pestle and mortar . The crushed bones are then chopped and incubated for 1 h at 37 °C in 20 ml of DMEM containing 0.2 % ( wt/vol ) collagenase . The cell suspension is filtered before red blood cell lysis and incubated with the following antibodies : allophycocyanin ( P25054 ) -conjugated P09619 -α , FITC-conjugated Sca-1 , phycoerythrin ( PE ) -conjugated P08575 and Ter119 . Appropriate gates are constructed on a cell sorter to exclude dead cells and lineage ( P08575 (+)Ter-119(+) ) -positive cells . Approximately 10,000 PαS MSCs may then be isolated per mouse . The total protocol takes ~7 h to complete . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Exploring schizophrenia drug-gene interactions through molecular network and pathway modeling . In this study , we retrieved 39 schizophrenia-related antipsychotic drugs from the DrugBank database . These drugs had interactions with 142 targets , whose corresponding genes were defined as drug targeted genes . To explore the complexity between these drugs and their related genes in schizophrenia , we constructed a drug-target gene network . These genes were overrepresented in several pathways including : neuroactive ligand-receptor pathways , glutamate metabolism , and glycine metabolism . Through integrating the pathway information into a drug-gene network , we revealed a few bridge genes connected the sub-networks of the drug-gene network : Q12879 , O60391 , Q14957 , Q13224 , P21728 , and P14416 . These genes encode ionotropic glutamate receptors belonging to the DB01221 receptor family and dopamine receptors . DB00502 was the only drug to directly interact with these pathways and receptors and consequently may have a unique action at the drug-gene interaction level during the treatment of schizophrenia . This study represents the first systematic investigation of drug-gene interactions in psychosis . Kinin-B2 receptor exerted neuroprotection after diisopropylfluorophosphate-induced neuronal damage . The kinin-B2 receptor ( B2BKR ) activated by its endogenous ligand bradykinin participates in various metabolic processes including the control of arterial pressure and inflammation . Recently , functions for this receptor in brain development and protection against glutamate-provoked excitotoxicity have been proposed . Here , we report neuroprotective properties for bradykinin against organophosphate poisoning using acute hippocampal slices as an in vitro model . Following slice perfusion for 10min with diisopropylfluorophosphate ( DB00677 ) to initiate the noxious stimulus , responses of pyramidal neurons upon an electric impulse were reduced to less than 30 % of control amplitudes . Effects on synaptic-elicited population spikes were reverted when preparations had been exposed to bradykinin 30min after challenging with DB00677 . Accordingly , bradykinin-induced population spike recovery was abolished by HOE-140 , a B2BKR antagonist . However , the kinin-B1 receptor ( B1BKR ) agonist Lys-des- DB00125 (9)-bradykinin , inducing the phosphorylation of mitogen-activated protein kinase ( MEK/MAPK ) and cell death , abolished bradykinin-mediated neuroprotection , an effect , which was reverted by the P29323 inhibitor PD98059 . In agreement with pivotal B1BKR functions in this process , antagonism of endogenous B1BKR activity alone was enough for restoring population spike activity . On the other hand pralidoxime , an oxime , reactivating acetylcholinesterase ( P22303 ) after organophosphate poisoning , induced population spike recovery after DB00677 exposure in the presence of bradykinin and Lys-des- DB00125 (9)-bradykinin . Lys-des- DB00125 (9)-bradykinin did not revert protection exerted by pralidoxime , however when instead bradykinin and Ly-des- DB00125 (9)-bradykinin were superfused together , recovery of population spikes diminished . These findings again confirm the neuroprotective feature of bradykinin , which is , diminished by its endogenous metabolites , stimulating the B1BKR , providing a novel understanding of the physiological roles of these receptors . DB08896 in metastatic colorectal cancer . DB08896 is an oral multikinase inhibitor that blocks the activity of protein kinases involved in the regulation of tumor angiogenesis ( P17948 , 2 , 3 ; angiopoietin-1 receptor ) , oncogenesis ( stem cell growth factor receptor ; P07949 ; P15056 including BRAFV600E ) , and tumor microenvironment ( P09619 -β and FGFR ) . Based on data from the Phase III CORRECT study , regorafenib stands as a further option for patient affected by metastatic colorectal cancer who have exhausted previous available therapies . Its multi-targeted effect might explain activity in advanced lines of treatment , when cancer cells have been heavily challenged with previous lines of therapy and potentially developed multiple mechanisms of resistance , but also makes difficult to identify predictive biomarkers . In this article we examine preclinical as well as clinical data of regorafenib in the therapy of metastatic colorectal cancer , challenges for potential markers of efficacy and its role in the treatment algorithm . Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 -I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 -Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 -I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 -Is . Treatment for the toxicity of P22303 -Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 -Is exert their effects . By attenuating the effects of P22303 -Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 -I . Timing of the hepatic arterial phase at DB08884 -enhanced hepatic dynamic Q9BWK5 : comparison of the test-injection and the fixed-time delay method . PURPOSE : To compare the fixed-time- and the test-injection method with respect to the image quality of hypervascular hepatocellular carcinoma ( HCC ) and the adequacy of timing of the hepatic arterial phase ( HAP ) in DB08884 ( EOB ) enhanced Q9BWK5 . MATERIALS AND METHODS : We studied 63 patients with computed tomography ( CT ) -proven hypervascular HCC : 30 ( group 1 ) were scanned HAP using the fixed-time delay method ( protocol 1 ) ; in the other 33 ( group 2 ) , we applied the test-injection method ( protocol 2 ) . We compared the protocols with respect with tumor-to-liver contrast ( TLC ) , contrast-to-noise-ratio ( P21554 ) , and relative enhancement of the liver and tumor ( Q04864 , P07949 ) during HAP . Two radiologists compared the adequacy of HAP , image contrast , image noise , and overall image quality . RESULTS : Under protocol 2 , TLC , P21554 , and Q04864 and P07949 of hypervascular HCC were significantly higher ( P < 0.01 ) . The proportion of optimal HAP was significantly higher for protocol 2 than protocol 1 ( P < 0.01 ) . The visual score of the image contrast and the overall image quality were significantly higher in group 2 than group 1 ( P = 0.02 and P = 0.01 , respectively ) . CONCLUSION : At EOB-enhanced hepatic dynamic Q9BWK5 , the test-injection method yielded better image quality of hypervascular HCC and improved adequacy of timing of HAP . Nerve growth factor activation of the extracellular signal-regulated kinase pathway is modulated by Ca(2+) and calmodulin . Nerve growth factor is a member of the neurotrophin family of trophic factors that have been reported to be essential for the survival and development of sympathetic neurons and a subset of sensory neurons . Nerve growth factor exerts its effects mainly by interaction with the specific receptor TrkA , which leads to the activation of several intracellular signaling pathways . Once activated , TrkA also allows for a rapid and moderate increase in intracellular calcium levels , which would contribute to the effects triggered by nerve growth factor in neurons . In this report , we analyzed the relationship of calcium to the activation of the Ras/extracellular signal-regulated kinase pathway in PC12 cells . We observed that calcium and calmodulin are both necessary for the acute activation of extracellular signal-regulated kinases after TrkA stimulation . We analyzed the elements of the pathway that lead to this activation , and we observed that calmodulin antagonists completely block the initial P04049 activation without affecting the function of upstream elements , such as Ras , Grb2 , Shc , and Trk . We have broadened our study to other stimuli that activate extracellular signal-regulated kinases through tyrosine kinase receptors , and we have observed that calmodulin also modulates the activation of such kinases after epidermal growth factor receptor stimulation in PC12 cells and after TrkB stimulation in cultured chicken embryo motoneurons . P62158 seems to regulate the full activation of P04049 after Ras activation , since functional Ras is necessary for P04049 activation after nerve growth factor stimulation and calmodulin-Sepharose is able to precipitate P04049 in a calcium-dependent manner . DB04868 and MEK inhibitors induce synthetic lethality through paradoxical activation of RAF in drug-resistant chronic myeloid leukemia . We show that imatinib , nilotinib , and dasatinib possess weak off-target activity against RAF and , therefore , drive paradoxical activation of P15056 and CRAF in a DB01367 -dependent manner . Critically , because DB01367 is activated by P11274 - P00519 , in drug-resistant chronic myeloid leukemia ( CML ) cells , DB01367 activity persists in the presence of these drugs , driving paradoxical activation of P15056 , CRAF , MEK , and P29323 , and leading to an unexpected dependency on the pathway . Consequently , nilotinib synergizes with MEK inhibitors to kill drug-resistant CML cells and block tumor growth in mice . Thus , we show that imatinib , nilotinib , and dasatinib drive paradoxical RAF/MEK/ P29323 pathway activation and have uncovered a synthetic lethal interaction that can be used to kill drug-resistant CML cells in vitro and in vivo . DB08896 . DB08896 ( DB08896 , Stivarga® ) is an oral diphenylurea multikinase inhibitor that targets angiogenic ( P17948 -3 , Q02763 ) , stromal ( P09619 -β , FGFR ) , and oncogenic receptor tyrosine kinases ( P10721 , P07949 , and RAF ) . DB08896 is the first small-molecule multikinase inhibitor to achieve survival benefits in metastatic colorectal cancer that has progressed after all standard therapies . Consequently , regorafenib was FDA approved for this indication . In addition , regorafenib treatment resulted in a significant improvement in progression-free survival ( PFS ) compared with placebo in patients with metastatic gastrointestinal stromal tumors ( GIST ) after progression on standard treatments and is also an FDA approved indication . Currently , regorafenib is examined in several clinical trials ( mostly phase II ) in different tumor entities , including renal cell carcinoma ( RCC ) , hepatocellular carcinoma ( HCC ) , and soft tissue sarcoma ( STS ) . The development of regorafenib and its current and potential future role in cancer therapy . DB08896 is a novel multikinase inhibitor that has demonstrated broad antitumor activity across various solid tumor types , in preclinical and clinical studies . Preclinical data show inhibitory activity of angiogenic , stromal and oncogenic tyrosine kinases through the targeting of vascular endothelial growth factor receptors 1 , 2 and 3 , tyrosine-protein kinase receptor P35590 -2 , platelet-derived growth factor receptor β , fibroblast growth factor receptor 1 , proto-oncogene tyrosine-protein kinase receptor Ret , mast/stem cell growth factor receptor Kit , P04049 and wild-type and V600E mutant serine/threonine-protein kinase B-raf . Phase I trials have shown that the drug is relatively well tolerated at doses of 160 mg daily on a 3-weeks-on/1-week-off schedule , or 100 mg daily on a continuous schedule , with adverse effects typical of other multikinase inhibitors . Phase II studies demonstrated clinical benefit in a variety of tumor types , mostly associated with prolonged stable disease . Phase III studies include the CORRECT trial , which ultimately led to FDA approval of the drug in the setting of metastatic colorectal cancer previously treated with standard therapies . There is still much work to be done to determine the role of regorafenib in the future of cancer therapy . This review will focus on the development of regorafenib , from early preclinical work through phase I , II and III trials , as well as highlighting the current role and potential future directions of this novel agent . Analyses of cross species polymerase chain reaction products to infer the ancestral state of human polymorphisms . In numerous population genetic and disease association studies decisions about the ancestry of polymorphic alleles are often made based on the relative frequency of the alleles in the extant populations with the most frequent allele being deemed as ancestral . However , the frequency of an allele in a population is generally not a perfect indicator of its ancestral status . A more accurate method to assess ancestral/derived status of polymorphic alleles involves identification of shared alleles between species . We used this strategy to examine genomic regions homologous to several human polymorphisms in four species of non-human primates . Cross species polymerase chain reaction ( CS-PCR ) , with primers designed from human sequence , was used to investigate regions of interest . Nineteen polymorphisms at six loci ( P14416 , HOXB@ , PAH , D4S10 , P10745 , and P07949 ) were examined either by restriction fragment length analysis of PCR products ( PCR-RFLP ) or by direct sequencing . At seventeen of the eighteen PCR-RFLPs , non-human primates were monomorphic and identical to each other for either lack of restriction enzyme site or presence of the site . Thus , at these seventeen polymorphic sites the shared alleles are most likely to be the ancestral ones in humans . In several cases we have used sequence data to further demonstrate that the nucleotide at the site of the polymorphism is conserved between species confirming the hypothesis of a single ancestral allele . However , not all human alleles can be simply resolved into ancestral and derived ; sequence data from one PCR-RFLP ( in an intron of the PAH locus ) and a single strand conformational polymorphism ( SSCP ) in the 3' untranslated region ( UTR ) of the P14416 gene illustrate this point . Oncogenes in melanoma : an update . Melanoma is a highly aggressive tumour with poor prognosis in the metastatic stage . P15056 , P01111 , and P10721 are three well-known oncogenes involved in melanoma pathogenesis . Targeting of mutated P15056 kinase has recently been shown to significantly improve overall survival of metastatic melanoma patients , underscoring the particular role of this oncogene in melanoma biology . However , recurrences regularly occur within several months , which supposedly involve further oncogenes . Moreover , oncogenic driver mutations have not been described for up to 30 % of all melanomas . In order to obtain a more complete picture of the mutational landscape of melanoma , more recent studies used high-throughput DNA sequencing technologies . A number of new oncogene candidates such as P28482 /2 , Q15303 , Q12879 , Q14832 , P63000 , and Q70Z35 were identified . Their particular role in melanoma biology is currently under investigation . Evidence for the functional relevance of some of these new oncogene candidates has been provided in in vitro and in vivo experiments . However , these findings await further validation in clinical studies . This review provides an overview on well-known melanoma oncogenes and new oncogene candidates , based on recent high-throughput sequencing studies . The list of genes discussed herein is of course not complete but highlights some of the most significant of recent findings in this area . The new candidates may support more individualized treatment approaches for metastatic melanoma patients in the future . The role of HIV-related chemokine receptors and chemokines in human erythropoiesis in vitro . In order to better define the role of HIV-related chemokines in human erythropoiesis we studied : A ) the expression of chemokine receptors , both on human P28906 (+) cells which include erythroid progenitors and on more mature erythroid cells ; B ) the functionality of these receptors by calcium flux , chemotaxis assay and phosphorylation of mitogen-activated protein kinases ( MAPK ) Q8NFH3 /44 ( P27361 / P28482 ) and AKT , and finally C ) the influence of chemokines on BFU-E formation . We found that HIV-related chemokine receptor P61073 , but not P51681 , is detectable on human P28906 (+) BFU-E cells . P61073 surface expression decreased during erythroid maturation , although P61073 mRNA was still present in cells isolated from differentiated erythroid colonies . P48061 , a P61073 ligand , induced calcium flux and phosphorylation of MAPK ( Q8NFH3 /44 ) and AKT in P28906 (+) P10721 (+) bone marrow mononuclear cells which contain BFU-E , as well as chemotactic activity of both human P28906 (+) BFU-E progenitors and erythroid cells isolated from day 2-6 BFU-E colonies . Responsiveness to P48061 decreased when the cells differentiated to the point of surface expression of the erythroid-specific marker P02724 . In contrast , the P51681 ligands ( macrophage inflammatory protein-1alpha [ MIP-1alpha ] , MIP-1beta , and RANTES ) did not activate calcium flux , MAPK and AKT phosphorylation or chemotaxis of P28906 (+) P10721 (+) cells or cells isolated from the BFU-E colonies . Interestingly , none of the chemokines tested in this study had any effect on BFU-E colony formation . In conclusion , only P61073 is functional , and its specific ligand P48061 may therefore play an important role in the homing and/or retention of early erythroid precursors in the bone marrow environment . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . DB08896 for cancer . INTRODUCTION : DB08896 ( DB08896 ) is a novel , orally active , diphenylurea multikinase inhibitor of P17948 -3 , c- P10721 , P35590 -2 , P09619 -β , P11362 , P07949 , RAF-1 , P15056 and p38 Q96HU1 kinase . AREAS COVERED : This review covers the preclinical development of regorafenib as well as the pivotal Phase I studies . The safety profile of regorafenib is discussed in context with other oral multikinase inhibitors bearing a similar target profile . Current clinical developments , especially in colorectal cancer ( CRC ) and gastrointestinal stromal tumor ( GIST ) , are addressed . Open questions on clinically useful biomarkers predicting response with regard to a personalized therapy strategy are also being discussed . EXPERT OPINION : DB08896 ( DB08896 ) is a novel , orally active multikinase inhibitor that is well tolerated in preclinical mouse models as well as clinically according to Phase I - III trials performed . The toxicity profile is comparable with other oral multikinase inhibitors with similar molecular targets . DB08896 has promising antineoplastic activity in various tumor types . Two large , randomized Phase III pivotal registration studies in patients with GIST and CRC , respectively , already completed enrolment , with final results being awaited . Further extensive clinical development as a single agent or in combination with standard chemotherapeutic agents in various malignant tumors is ongoing . Moreover , regorafenib has recently been granted Orphan Drug Status for GIST tumors and ' fast track ' status for both GIST and CRC by the FDA . P14416 signaling dynamics of dopamine D2-neurotensin 1 receptor heteromers . Biochemical , histochemical and coimmunoprecipitation experiments have indicated the existence of antagonistic dopamine D2 ( D2R ) and neurotensin 1 ( NTS1R ) receptor-receptor interactions in the dorsal and ventral striatum indicating a potential role of these receptor-receptor interactions in Parkinson 's disease and schizophrenia . By means of Bioluminiscence Resonance energy transfer ( BRET(2) ) evidence has for the first time been obtained in the current study for the existence of both D2LR/NTS1R and D2SR/NTS1R heteromers in living HEK293T cells . Through confocal laser microscopy the NTS1R(GFP2) and D2R(YFP) were also shown to be colocated in the plasma membrane of these cells . A bioinformatic analysis suggests the existence of a basic set of three homology protriplets ( TVM , DLL and/or LRA ) in the two participating receptors which may contribute to the formation of the D2R/NTS1R heteromers by participating in guide-clasp interactions in the receptor interface . The CREB reporter gene assay indicated that the neurotensin receptor agonist JMV 449 markedly reduced the potency of the D2R like agonist quinpirole to inhibit the forskolin induced increase of the CREB signal . In contrast , the neurotensin agonist was found to markedly increase the quinpirole potency to activate the MAPK pathway as also studied with luciferase reporter gene assay measuring the degree of SRE activity as well as with P27361 /2 phosphorylation assays . These dynamic changes in D2R signaling produced by the neurotensin receptor agonist may involve antagonistic allosteric receptor-receptor interactions in the D2LR-NTS1R heteromers at the plasma membrane level ( CREB pathway ) and synergistic interactions in PKC activation at the cytoplasmatic level ( MAPK pathway ) . Association of DNA repair gene polymorphisms with response to platinum-based doublet chemotherapy in patients with non-small-cell lung cancer . PURPOSE : To identify polymorphisms in DNA repair genes that affect responses to platinum-based doublet chemotherapy in patients with non-small-cell lung cancer ( NSCLC ) . PATIENTS AND METHODS : In total , 640 patients with NSCLC who received platinum-based doublet chemotherapy in the National Cancer Center Hospital in Japan from 2000 to 2008 and whose responses were evaluated by Response Evaluation Criteria in Solid Tumors ( RECIST ) participated in a study of the association between response and genotypes for 30 single nucleotide polymorphisms ( SNPs ) in 27 DNA repair genes . Candidate SNPs were selected in a discovery set of 201 patients , and their associations were validated in an independent set of 439 patients by prespecified P value criteria . RESULTS : Homozygotes for the minor allele P04637 -72Pro of the Arg72Pro SNP in the P04637 gene showed a better response rate ( 54.3 % ) than those for the major allele P04637 -72Arg ( 29.1 % ; P = 4.4 × 10(-5) ) irrespective of therapeutic regimens , and minor allele homozygotes had significantly longer progression-free and overall survivals than major allele homozygotes ( hazard ratio [ HR ] , 0.85 ; 95 % CI , 0.74 to 0.98 ; P = .020 ; and HR , 0.86 ; 95 % CI , 0.74 to 0.99 ; P = .039 ) . Minor allele carriers for SNP Lys940Arg in the poly ( ADP-ribose ) polymerase 1 ( P09874 ) gene showed a better response rate to the paclitaxel regimen ( 45.8 % ) than to the gemcitabine regimen ( 10.5 % ; P for interaction = .019 ) . CONCLUSION : Polymorphisms in the P04637 and P09874 genes are involved in inter-individual differences in the response to platinum-based doublet chemotherapy in patients with NSCLC . Molecular profiling in the treatment of colorectal cancer : focus on regorafenib . Metastatic colorectal cancer ( mCRC ) is a highly heterogeneous disease . Its treatment outcome has been significantly improved over the last decade with the incorporation of biological targeted therapies , including anti- P00533 antibodies , cetuximab and panitumumab , and P15692 inhibitors , bevacizumab , ramucirumab , and aflibercept . The identification of predictive biomarkers has further improved the survival by accurately selecting patients who are most likely to benefit from these treatments , such as DB01367 mutation profiling for P00533 antibodies . DB08896 is a multikinase inhibitor currently used as late line therapy for mCRC . The molecular and genetic markers associated with regorafenib treatment response are yet to be characterized . Here , we review currently available clinical evidence of mCRC molecular profiling , such as DB01367 , P15056 , and P22897 testing , and its role in targeted therapies with special focus on regorafenib treatment . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . DB04868 : a phenylamino-pyrimidine derivative with activity against P11274 - P00519 , P10721 and P09619 kinases . The P11274 - P00519 kinase inhibitor imatinib mesylate is currently the standard therapy for patients with chronic myeloid leukemia ( CML ) . However , mutations within the P00519 kinase domain interfering with drug binding have been identified as the main mechanism of resistance to imatinib . Multiple distinct P11274 - P00519 kinase mutant isoforms conferring varying degrees of resistance to tyrosine kinase inhibitors have been reported . DB04868 is a tyrosine kinase inhibitor 30-fold more potent than imatinib against P11274 - P00519 kinase . DB04868 is active against a wide range of imatinib-resistant P11274 - P00519 mutant isoforms , except for T315I . Results from Phase II studies of nilotinib for patients with CML after failure or intolerance to imatinib therapy have shown a favorable toxicity profile and confirmed the high efficacy of nilotinib in this setting . Studies addressing the activity of nilotinib in newly-diagnosed patients with CML are underway . Furthermore , nilotinib is a potent inhibitor of P10721 and P09619 kinases . Here , we review the preclinical development of nilotinib and the activity of this agent in patients with CML and in tumors driven by P10721 and/or P09619 mutant kinases , such as gastrointestinal stromal tumors and some forms of clonal hypereosinophilia . DB08896 for treatment of advanced gastrointestinal stromal tumors . INTRODUCTION : Gastrointestinal stromal tumors ( GISTs ) are abdominal sarcomas which are extremely refractory to chemotherapy treatment . The treatment of GISTs has been revolutionized by use of P10721 /platelet-derived growth factor receptor-α ( P16234 ) kinase inhibitors . Unfortunately , most tumors develop resistance to front-line ( imatinib ) or second-line ( sunitinib ) therapy . DB08896 , a P10721 / P16234 /vascular endothelial growth factor receptor ( VEGFR ) oral kinase inhibitor , has been shown to improve progression-free survival in the third- or fourth-line setting . AREAS COVERED : This review covers the preclinical and clinical studies of regorafenib for treatment of GIST . A literature search on regorafenib was carried out using the PubMed database up to October 2013 . EXPERT OPINION : Currently , imatinib and sunitinib represent the only proven first- and second-line therapies , respectively , for advanced GISTs . Based on the results of a Phase III study , regorafenib is now established as the only proven third-line therapy . DB08896 activity in this setting is believed to be due to its activity against oncogenic forms of P10721 / P16234 . Although side effects are common with this agent , they can be effectively managed with a combination of supportive care , dose interruptions/reductions . The toxicity profile is similar to other oral kinase inhibitors with anti-VEGFR activity . DB08896 is mainly metabolized by P08684 , and concomitant use of strong inducers/inhibitors of this enzyme should be avoided . Role of P15056 in thyroid oncogenesis . P15056 , a cytoplasmic serine-threonine protein kinase , plays a critical role in cell signaling as an activator within the mitogen-activated protein kinase ( MAPK ) pathway . The most common P15056 mutation is the V600E transversion , which causes constitutive kinase activity . This mutation has been found in a multitude of human cancers , including both papillary thyroid cancer ( PTC ) and papillary-derived anaplastic thyroid cancer ( ATC ) , in which it initiates follicular cell transformation . With such a high frequency of P15056 mutations in PTC ( 44 % ) and PTC-derived ATC ( 24 % ) , research in P15056 (V600E) detection for diagnostic purposes has shown high sensitivity and specificity for tumor cell presence . P15056 (V600E) in PTC has also provided valuable prognostic information , as its presence has been correlated with more aggressive and iodine-resistant phenotypes . Such findings have initiated research in targeting oncogenic P15056 in cancer therapeutics . Although multiple phase II clinical trials in patients with iodine-refractory metastatic PTC have shown significant efficacy for sorafenib , a first-generation P15056 inhibitor , the mechanism by which it mediates its effect remains unclear because of multiple additional kinase targets of sorafenib . Additionally , preclinical and clinical studies investigating combination therapy with agents such as selective ( PLX 4032 ) and potent ( DB08896 and ARQ 736 ) small-molecule P15056 inhibitors and Q96HU1 /extracellular signal-regulated kinase ( P29323 ) kinase inhibitors ( AZD6244 ) hold great promise in the treatment of P15056 (V600E) cancers and may eventually play a powerful role in changing the clinical course of PTC and ATC . Management of regorafenib-related toxicities : a review . DB08896 ( Stivarga , DB08896 ; Bayer Pharma AG , Berlin , Germany ) is an oral multikinase inhibitor that targets the angiogenic tumor microenvironment and oncogenic kinases including vascular endothelial growth factor receptor 2 ( P35968 ) , P17948 , P35916 , fibroblast growth factor receptor 1 ( P11362 ) , RAF , P10721 , P07949 and P15056 . Its antiangiogenic effect is greater than that of its related drug , sorafenib . DB08896 has been approved by the US Food and Drug Administration ( FDA ) for the treatment of metastatic colorectal cancer ( mCRC ) in patients who have failed treatment with fluoropyrimidine , oxaliplatin and irinotecan based chemotherapy , an anti- P15692 therapy and , if P01116 wild type , an anti- P00533 therapy . The FDA based this approval on data from the CORRECT trial , which showed the efficacy of regorafenib compared with placebo . The most common grade 3-4 adverse reactions with the drug are hand foot skin reactions ( HFSR ) , diarrhea , hypertension and fatigue . This review discusses the efficacy data , and the incidence and management of regorafenib 's toxicities . Poly( DB02059 )polymerase-1 signalling of the DNA damage induced by P11387 poison in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 inhibitor DB01030 and the poly( DB02059 )polymerase-1 inhibitor DB02690 in D54(p53wt) and U251(p53mut) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2/M block of the cell cycle . We also evaluated , the influence of TPT+/- DB02690 treatment on P09874 and p53 activity . We got evidences of a TPT-dependent increase of P09874 auto-modification level in both the cells . Moreover , in the D54(p53wt) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 nuclear amount . Conversely , in U251(p53mut) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 proteolysis . Our findings suggest that the modulation of P09874 can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status . Molecular-based choice of cancer therapy : realities and expectations . Current choice of cancer therapy is usually empirical and relies mainly on the statistical prediction of the treatment success . Molecular research provides some opportunities to personalize antitumor treatment . For example , life-threatening toxic reactions can be avoided by the identification of subjects , who carry susceptible genotypes of drug-metabolizing genes ( e.g. P51580 , P22309 , P42898 , Q12882 ) . Tumor sensitivity can be predicted by molecular portraying of targets and other molecules associated with drug response . Tailoring of antiestrogen and trastuzumab therapy based on hormone and P04626 receptor status has already become a classical example of customized medicine . Other predictive markers have been identified both for cytotoxic and for targeted therapies , and include , for example , expression of TS , TP , Q12882 , OPRT , P07992 , P16455 , P11388 , class III beta-tubulin molecules as well as genomic alterations of P00533 , P10721 , P00519 oncogenes . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . AM251 induces apoptosis and G2/M cell cycle arrest in A375 human melanoma cells . Human cutaneous melanoma is an aggressive and chemotherapy-resistant type of cancer . AM251 is a cannabinoid type 1 ( P21554 ) receptor antagonist/inverse agonist with off-target antitumor activity against pancreatic and colon cancer cells . The current study aimed to characterize the in-vitro antimelanoma activity of AM251 . The P15056 V600E mutant melanoma cell line , A375 , was used as an in-vitro model system . Characterization tools included a cell viability assay , nuclear morphology assessment , gene expression , western blot , flow cytometry with P08758 -FITC/7-AAD double staining , cell cycle analyses , and measurements of changes in intracellular DB02527 and calcium concentrations . AM251 exerted a marked cytotoxic effect against A375 human melanoma cells with potency comparable with that observed for cisplatin without significant changes in the human dermal fibroblasts viability . AM251 , at a concentration that approximates the IC50 , downregulated genes encoding antiapoptotic proteins ( P10415 and survivin ) and increased transcription levels of proapoptotic Q07812 , induced alteration of P08758 reactivity , DNA fragmentation , chromatin condensation in the cell nuclei , and G2/M phase arrest.AM251 also induced a 40 % increase in the basal DB02527 levels , but it did not affect intracellular calcium concentrations . The involvement of Q9Y2T6 , O75762 , and P35354 in the AM251 mechanism of action was excluded . The combination of AM251 with celecoxib produced a synergistic antitumor activity , although the mechanism underlying this effect remains to be elucidated . This study provides the first evidence of a proapoptotic effect and G2/M cell cycle arrest of AM251 on A375 cells . This compound may be a potential prototype for the development of promising diarylpyrazole derivatives to be evaluated in human cutaneous melanoma . Order of genes on human chromosome 5q with respect to 5q interstitial deletions . Using ( a ) somatic cell hybrids retaining partial chromosome 5 and ( b ) clinical samples from patients with acquired deletions of the long arm of chromosome 5 , combined with chromosome 5-linked DNA probes , some of which exhibited RFLPs , we have determined the order of a series of genes on chromosome 5 . The order established is 5pter ---- MLVI-2 ---- cen ---- P07686 ---- P00374 ---- Pi227- --- cp12.6 ---- ( P05113 , P05112 ) ---- P08700 ---- P04141 ---- P05230 ---- ( P07333 , P09619 ) ---- ( treC,ADRBR ) ---- ( Q5SW96 - Q13585 , P09603 ) ---- qter . The suggested order and orientation for the closely linked P08700 / P04141 gene pair is cen ---- 5' P08700 3' ---- 5' P04141 3' ---- qter , on the basis of analysis of the P04141 rearrangement in HL60 DNA . The map position of the GRL locus , which was consistent with both somatic cell hybrid and 5q- analyses , was telomeric to P04141 and centromeric to P07333 / P09619 , near P05230 . Long-range restriction-enzyme analysis of 5q- DNAs did not detect rearrangements of 5q-linked probes except in HL60 DNA , but it did reveal putative long-range RFLPs of several loci . RFLPs for GRL , Pi227 , cp12.6 , P08700 , and P07333 can detect deletions in bone marrow and in leukemia cells from patients with acquired 5q deletions . G protein-coupled receptor kinase-3-deficient mice exhibit WHIM syndrome features and attenuated inflammatory responses . Chemokine receptor interactions coordinate leukocyte migration in inflammation . Chemokine receptors are GPCRs that when activated , are phosphorylated by GRKs to turn off G protein-mediated signaling yet recruit additional signaling machinery . Recently , P35626 was identified as a negative regulator of P48061 / P61073 signaling that is defective in human WHIM syndrome . Here , we report that P35626 -/- mice exhibit numerous features of human WHIM , such as impaired P48061 -mediated desensitization , enhanced P61073 signaling to P29323 activation , altered granulocyte migration , and a mild myelokathexis . Moreover , P35626 -/- protects mice from two acute models of inflammatory arthritis ( K/BxN serum transfer and CAIA ) . In these granulocyte-dependent disease models , protection of P35626 -/- mice is mediated by retention of cells in the marrow , fewer circulating granulocytes in the peripheral blood , and reduced granulocytes in the joints during active inflammation . In contrast to WHIM , P35626 -/- mice have minimal hypogammaglobulinemia and a peripheral leukocytosis with increased lymphocytes and absent neutropenia . Thus , we conclude that the loss of P35626 -mediated regulation of P48061 / P61073 signaling contributes to some , but not all , of the complete WHIM phenotype and that P35626 inhibition may be beneficial in the treatment of inflammatory arthritis . N-arachidonoyl-L-serine is neuroprotective after traumatic brain injury by reducing apoptosis . N-arachidonoyl-L-serine ( AraS ) is a brain component structurally related to the endocannabinoid family . We investigated the neuroprotective effects of AraS following closed head injury induced by weight drop onto the exposed fronto-parietal skull and the mechanisms involved . A single injection of AraS following injury led to a significant improvement in functional outcome , and to reduced edema and lesion volume compared with vehicle . Specific antagonists to CB2 receptors , transient receptor potential vanilloid 1 ( Q8NER1 ) or large conductance calcium-activated potassium ( BK ) channels reversed these effects . Specific binding assays did not indicate binding of AraS to the Q9Y2T6 cannabinoid receptor . N-arachidonoyl-L-serine blocked the attenuation in phosphorylated extracellular-signal-regulated kinase 1/2 ( P29323 ) levels and led to an increase in pAkt in both the ipsilateral and contralateral cortices . Increased levels of the prosurvival factor Bcl-xL were evident 24 hours after injury in AraS-treated mice , followed by a 30 % reduction in caspase-3 activity , measured 3 days after injury . Treatment with a CB2 antagonist , but not with a P21554 antagonist , reversed this effect . Our results suggest that administration of AraS leads to neuroprotection via P29323 and Akt phosphorylation and induction of their downstream antiapoptotic pathways . These protective effects are related mostly to indirect signaling via the CB2R and Q8NER1 channels but not through P21554 or Q9Y2T6 receptors . Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . Glucocorticoid-induced surface expression of annexin 1 blocks beta2-integrin adhesion of human eosinophils to intercellular adhesion molecule 1 surrogate protein . BACKGROUND : Glucocorticoids attenuate the population of eosinophils and T lymphocytes in asthmatic airways . The decrease in airway eosinophilia is caused both by accelerated cell death and by induction of blockade of integrin adhesion . In this study , we examined the hypothesis that annexin 1 surface expression , which is upregulated by the glucocorticoid receptor , prevents integrin adhesion essential to cell migration by blocking intracellular translocation of cytosolic group IV phospholipase A2 ( P47712 ) . OBJECTIVE : To examine the relationship of the glucocorticoid on annexin 1 expression and the effect of blockade of annexin 1 activity on adhesion of human eosinophils in vitro . To determine the relationship between annexin 1surface expression and nuclear membrane translocation of P47712 . METHODS : Eosinophils isolated from human peripheral blood were pretreated with fluticasone propionate ( FP ) , and beta2-integrin adhesion was measured after stimulation with P05113 or eotaxin . Effects of FP on P47712 expression , phosphorylation , and translocation were determined . The role of annexin 1 was examined by using annexin 1 blocking antibody and/or mimetic peptides . RESULTS : DB00588 decreased stimulated eosinophil adhesion and caused 4-fold increase in annexin 1 expression on the plasma membrane . Inhibition of adhesion by FP was blocked with annexin 1 blocking antibody . Annexin 1 N-terminal mimetic peptide also blocked beta2-integrin adhesion . Translocation of P47712 to the nuclear membrane was significantly blocked by incubation with FP . Blockade was reversed with annexin 1 blocking antibody . CONCLUSION : Blockade of beta2-integrin adhesion by glucocorticoid is regulated by annexin 1 , which blocks P47712 translocation to nuclear membrane . P48061 and [N33A] P48061 in 5637 and HeLa cells : regulating P00533 phosphorylation via calmodulin/calcineurin . In the human neoplastic cell lines 5637 and HeLa , recombinant P48061 elicited , as expected , downstream signals via both G-protein-dependent and β-arrestin-dependent pathways responsible for inducing a rapid and a late wave , respectively , of P27361 /2 phosphorylation . In contrast , the structural variant [N33A] P48061 triggered no β-arrestin-dependent phosphorylation of P27361 /2 , and signaled via G protein-dependent pathways alone . Both P48061 and [N33A] P48061 , however , generated signals that transinhibited P00533 phosphorylation via intracellular pathways . 1 ) Prestimulation of P61073 / P00533 -positive 5637 or HeLa cells with P48061 modified the HB- P01133 -dependent activation of P00533 by delaying the peak phosphorylation of tyrosine 1068 or 1173 . 2 ) Prestimulation with the synthetic variant [N33A] P48061 , while preserving P61073 -related chemotaxis and P61073 internalization , abolished P00533 phosphorylation . 3 ) In cells knockdown of β-arrestin 2 , P48061 induced a full inhibition of P00533 like [N33A] P48061 in non-silenced cells . 4 ) P00533 phosphorylation was restored as usual by inhibiting PCK , calmodulin or calcineurin , whereas the inhibition of CaMKII had no discernable effect . We conclude that both recombinant P48061 and its structural variant [N33A] P48061 may transinhibit P00533 via G-proteins/calmodulin/calcineurin , but [N33A] P48061 does not activate β-arrestin-dependent P27361 /2 phosphorylation and retains a stronger inhibitory effect . Therefore , we demonstrated that P48061 may influence the magnitude and the persistence of signaling downstream of P00533 in turn involved in the proliferative potential of numerous epithelial cancer . In addition , we recognized that [N33A] P48061 activates preferentially G-protein-dependent pathways and is an inhibitor of P00533 . DB08896 ( DB08896 ) : a new oral multikinase inhibitor of angiogenic , stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity . Angiogenesis , a critical driver of tumor development , is controlled by interconnected signaling pathways . Vascular endothelial growth factor receptor ( VEGFR ) 2 and tyrosine kinase with immunoglobulin and epidermal growth factor homology domain 2 play crucial roles in the biology of normal and tumor vasculature . DB08896 ( DB08896 ) , a novel oral multikinase inhibitor , potently inhibits these endothelial cell kinases in biochemical and cellular kinase phosphorylation assays . Furthermore , regorafenib inhibits additional angiogenic kinases ( P17948 /3 , platelet-derived growth factor receptor-β and fibroblast growth factor receptor 1 ) and the mutant oncogenic kinases P10721 , P07949 and B-RAF . The antiangiogenic effect of regorafenib was demonstrated in vivo by dynamic contrast-enhanced magnetic resonance imaging . DB08896 administered once orally at 10 mg/kg significantly decreased the extravasation of Gadomer in the vasculature of rat GS9L glioblastoma tumor xenografts . In a daily (qd)×4 dosing study , the pharmacodynamic effects persisted for 48 hr after the last dosing and correlated with tumor growth inhibition ( TGI ) . A significant reduction in tumor microvessel area was observed in a human colorectal xenograft after qd×5 dosing at 10 and 30 mg/kg . DB08896 exhibited potent dose-dependent TGI in various preclinical human xenograft models in mice , with tumor shrinkages observed in breast MDA-MB-231 and renal 786-O carcinoma models . Pharmacodynamic analyses of the breast model revealed strong reduction in staining of proliferation marker Ki-67 and phosphorylated extracellular regulated kinases 1/2 . These data demonstrate that regorafenib is a well-tolerated , orally active multikinase inhibitor with a distinct target profile that may have therapeutic benefit in human malignancies . Complementary and alternative medicine use and quality of life in patients with primary brain tumors . This study explored the use of complementary and alternative medicine ( P62158 ) approaches and their relationship with demographic and disease characteristics and quality of life ( QOL ) in the primary brain tumor ( P10721 ) population . One hundred one P10721 patients were enrolled in this study . The results showed that 34 % of patients reported using P62158 . Forty-one percent reported using more than one type of P62158 . The average cost of each P62158 used per month was 69 dollars , with 20 % of patients spending more than 100 dollars per month . The majority ( 74 % ) reported that their physicians were unaware of their use of P62158 . Data analysis found a higher performance status to be the only factor significantly related to use of P62158 therapy ( P < 0.005 ) . There was no difference in patient report of QOL between users and nonusers of P62158 therapies . The high number of patients who do not report P62158 use has potential implications for evaluation of symptoms and response to therapy in this population . This may be especially relevant in those patients with higher functional status participating in clinical trials . Biological and molecular effects of small molecule kinase inhibitors on low-passage human colorectal cancer cell lines . Low-passage cancer cell lines are versatile tools to study tumor cell biology . Here , we have employed four such cell lines , established from primary tumors of colorectal cancer ( CRC ) patients , to evaluate effects of the small molecule kinase inhibitors ( SMI ) vemurafenib , trametinib , perifosine , and regorafenib in an in vitro setting . The mutant P15056 ( V600E/V600K ) inhibitor vemurafenib , but also the Q02750 /2 inhibitor trametinib efficiently inhibited DNA synthesis , signaling through P27361 /2 and expression of genes downstream of P27361 /2 in P15056 mutant cells only . In case of the AKT inhibitor perifosine , three cell lines showed a high or intermediate responsiveness to the drug while one cell line was resistant . The multikinase inhibitor regorafenib inhibited proliferation of all CRC lines with similar efficiency and independent of the presence or absence of P01116 , P15056 , P42336 , and P04637 mutations . DB08896 action was associated with broad-range inhibitory effects at the level of gene expression but not with a general inhibition of AKT or MEK/ P29323 signaling . In vemurafenib-sensitive cells , the antiproliferative effect of vemurafenib was enhanced by the other SMI . Together , our results provide insights into the determinants of SMI efficiencies in CRC cells and encourage the further use of low-passage CRC cell lines as preclinical models .

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Rationalizing cyclooxygenase ( P36551 ) inhibition for maximal efficacy and minimal adverse events . New information indicates that cyclooxygenase-2 ( P35354 ) is constitutively expressed in several tissues , including brain , lung , pancreas , kidney , and ovary , and plays an important role in renal and gastrointestinal function . Selective P35354 inhibition has been associated in animal studies with impairment of ulcer healing and renal function and inhibition of prostacyclin , an effect that inhibits vasodilation without inhibiting platelet aggregation . The clinical consequences , if any , of these effects remain to be determined in long-term studies in humans . The premise that selective P35354 inhibitors will cause less gastrointestinal toxicity than nonsteroidal antiinflammatory drugs that inhibit both P36551 isoforms needs to take into account the low toxicity of nabumetone . The gastrointestinal safety profile of this nonacidic , dual P36551 inhibitor that does not undergo enterohepatic circulation has been evaluated in extensive clinical trials . The data submitted to the US Food and Drug Administration in the New Drug Application for nabumetone ( DB00461 ) , the comparative trials subsequently completed , the published databases of the comparative gastrointestinal toxicity of various nonsteroidal anti-inflammatory drugs ( NSAIDs ) , and the meta-analysis published in this issue of The American Journal of Medicine ( Schoenfeld , page 48S ) indicate that nabumetone has the lowest incidence of gastrointestinal toxicity among the extensively studied NSAIDs . Overall , the incidence is approximately 10-fold less than with comparator drugs . This rate is an appropriate current reference against which the gastrointestinal toxicity of P35354 inhibitors can be compared . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Cytoplasmic and nuclear estrogen binding capacity in the rat uterus during treatment with danazol and testosterone . DB01406 , testosterone and dihydrotestosterone ( DB02901 ) were tested as competitors for estrogen receptors on immature rat uterus cytosol . No competitive binding could be demonstrated for any of these steroids . After that , prepubertal Wistar rats were exposed to danazol , testosterone or propylene glycol ( control ) for 3 days or 17 days . After the appropriate exposure to medication , the animals were killed . Both danazol and testosterone appeared to be uterotropic after 3 days of treatment , although the increase in the uterine weight was significant only in the danazol-treated group ( p less than 0.05 ) . This effect was lost after 17 days of treatment . P03372 binding assays were done on the cytosolic and nuclear fractions of the homogenized uterine tissue of each group . The estrogen binding capacity of cytosols was increased in both the danazol ( p less than 0.05 ) and the testosterone ( p less than 0.01 ) groups after 3 days of treatment . A parallel increase was found in the nuclear fraction of both groups . After 17 days of treatment , the comparison between the 3 groups showed no differences in the cytosolic or nuclear estrogen binding capacity . The information provided by this study suggests that some effects of danazol may be due to an androgenic action and that may be associated to increases in the free fraction of testosterone . Analysis of breast cancer related gene expression using natural splines and the Cox proportional hazard model to identify prognostic associations . Many studies correlating gene expression data to clinical parameters assume a linear increase or decrease of the clinical parameter under investigation with the expression of a gene . We have studied genes encoding important breast cancer-related proteins using a model for survival-type data that is based on natural splines and the Cox proportional hazard model , thereby removing the linearity assumption . Expression data of 16 genes were studied in relation to metastasis-free probability in a cohort of 295 consecutive breast cancer patients treated at The Netherlands Cancer Institute . The independent predictive power for disease outcome of the 16 individual genes was tested in a multivariable model with known clinical and pathological risk factors . There is a linear relationship between increasing expression and a higher or lower hazard for distant metastasis for P03372 , Q15303 , P15692 , O96020 , Q15910 , and Q96NZ9 ; for P04626 , P21860 , P24385 , P24864 , O75530 , P61073 , P32248 , P48061 , and P05121 there is no clear increase or decrease ; and for P00533 there seems to be a non-linear relation . Multivariable analysis showed that the 70-gene prognosis profile outperforms all the other variables in the model ( hazard-rate 5.4 , 95 % CI 2.5-11.7 ; P = 0.000018 ) . P00533 -expression seems to have a non-linear relation with disease outcome , indicating that lower but also higher expression of P00533 are associated with worse outcome compared to intermediate expression levels ; the other genes show no or a linear relation . Impact of 27-hydroxycholesterol on amyloid-beta peptide production and DB00171 -binding cassette transporter expression in primary human neurons . DB04540 is an integral component of neuronal membranes and recent evidence has shown that it regulates amyloid-beta protein precursor processing to form amyloid-beta peptides , which are a major constituent of cerebral amyloid plaques associated with Alzheimer 's disease . 27-Hydroxycholesterol ( 27OHC ) is synthesized from cholesterol via sterol 27-hydroxylase ( Q02318 ) in the brain and , unlike cholesterol , can cross into the brain through the blood brain barrier from the circulation . Previous studies point toward a potential role for 27OHC in the regulation of neuronal amyloid-beta peptide generation , however , this has not been investigated in primary human neurons . Here we show that 27OHC significantly reduced amyloid-beta peptide detected in cell culture supernatants from primary human neurons . We also show that 27OHC does not affect alpha- , beta- or gamma-secretase activity but does upregulate the liver X receptor ( LXR ) responsive genes O95477 , P45844 and P02649 . These data suggest that 27OHC-mediated reduction in extracellular amyloid-beta peptide levels is potentially due to its action as an LXR ligand . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . DB01645 potentiates the P01160 effect on a K(+)-conductance in P29320 -293 cells . P29320 -293 cells are known to reflect many features of the late distal tubule . Furthermore , they have the ability to release urodilatin , the structural analog to P01160 . RT-PCR was performed to test for the expression of natriuretic peptide receptors . While the mRNA for the human P01160 receptor ( P16066 , P16066 ) could be amplified , the P09543 -specific receptor P20594 ( P20594 ) and the receptor specific for guanylins , P25092 , could not be detected . In patch clamp experiments the effects of P01160 ( 10 nM ) on membrane voltage ( V(m) ) were monitored and P29320 -293 cells depolarized by 2.3 +/- 0.5 mV ( n=14 ) . In the presence of the P01133 receptor blocker genistein ( 10 microM ) the effect of P01160 was increased by 65 % to 3.9 +/- 0.8 mV ( n=14 ) . After removal of genistein the P01160 -mediated depolarization further increased by 147 % to 5.7 +/- 1.0 mV ( n=14 ) . P01160 given repetitively without genistein had no increasing depolarizing effect in P29320 -293 cells with time . The P01160 effect could be fully blocked by 1 mM Ba(2+) and by 1 microM of the specific PKG inhibitor KT5823 indicating that P01160 inhibits a K(+)-conductance via a cGMP-dependent protein kinase . DB01645 itself hyperpolarized the membrane voltage of P29320 -293 cells by -3.9 +/- 0.6 mV ( n=11 ) and this effect could also be fully blocked by Ba(2+) ( -0.3 +/- 0.1 mV , n=5 ) , indicating that genistein activates a K(+)-conductance which contributes significantly to the membrane potential of P29320 -293 cells . Coordinate transcriptional regulation of bile acid homeostasis and drug metabolism . Drugs and bile acids are taken up into hepatocytes by specialized transport proteins localized at the basolateral membrane , e.g. , organic anion transporting polypeptides . Following intracellular metabolism by cytochrome P450 ( CYP ) enzymes , drug metabolites are excreted into bile or urine via DB00171 -dependent multidrug resistance proteins ( P08183 and MRPs ) . Bile acids are excreted mainly via the bile salt export pump ( O95342 , O95342 ) . The genes coding for drug and bile acid transporters and CYP enzymes are regulated by a complex network of transcriptional cascades , notably by the ligand-activated nuclear receptors Q96RI1 , O75469 , and CAR and by the ligand-independent nuclear receptor HNF-4alpha . The bile acid synthesizing enzymes P22680 , Q9UNU6 , and Q02318 are subject to negative feedback regulation by bile acids , which is partly mediated through the transcriptional repressor Q15466 . The role of transcriptional cofactors , such as Q15788 and P20142 -1 , in mediating the gene-specific effects of individual nuclear receptors is becoming increasingly evident . Purification and characterization of mouse O15528 overproduced by an Escherichia coli system coexpressing molecular chaperonins GroEL/ES . The expression of mouse O15528 in Escherichia coli has been dramatically enhanced by coexpression of GroEL/ES . To reveal the enzymatic properties of O15528 , we measured its hydroxylation activity toward vitamin D3 and DB01436 ( 1alpha(OH)D3 ) in addition to the physiological substrate DB00146 . Surprisingly , O15528 converted vitamin D3 to 1alpha, DB00146 . Both 1alpha-hydroxylation activity toward vitamin D3 , and 25-hydroxylation activity toward 1alpha(OH)D3 were observed . The Km and Vmax values for 25-hydroxylation activity toward 1alpha(OH)D3 were estimated to be 1.7 microM and 0.51 mol/min/mol P450 , respectively , while those for 1alpha-hydroxylation activity toward DB00146 were 0.050 microM and 2.73 mol/min/mol P450 , respectively . Note that the substrate must be fixed in the opposite direction in the substrate-binding pocket of O15528 between 1alpha-hydroxylation and 25-hydroxylation . Based on these results and the fact that human Q02318 and Streptomyces CYP105A1 also convert vitamin D3 to 1alpha, DB00146 , 1alpha-hydroxylation , and 25-hydroxylation of vitamin D3 appear to be closely linked together . Preimplantation mouse blastocysts fail to express CYP genes required for estrogen biosynthesis . Implantation is initiated on day 4 in the mouse and on day 13 in the pig . The preimplantation pig blastocyst synthesizes steroid hormones , but whether preimplantation rodent embryos also have this ability has remained unresolved for the last two decades . In this study , the mRNAs encoding NADPH-cytochrome P450 reductase ( P450-reductase ) , adrenodoxin , lanosterol 14-demethylase P450 ( Q16850 ) , 17 alpha-hydroxylase P450 ( P05093 ) , cholesterol side-chain cleavage P450 ( P05108 ) , sterol 27-hydroxylase P450 ( Q02318 ) , and aromatase P450 ( P11511 ) were examined in day 4 mouse blastocysts ( day 1 = vaginal plug ) and in day 13 and 16 pig blastocysts using reverse transcription-polymerase chain reaction ( RT-PCR ) . In mouse blastocysts , mRNAs of P450-reductase , adrenodoxin , and Q16850 , but not P05093 , P05108 , Q02318 , and P11511 , were detected . In agreement with this finding , no aromatase protein could be detected by immunohistochemistry . By contrast , all these mRNAs were detected in the pig blastocyst . Furthermore , both the ovarian and placental types of aromatase ( P11511 ) mRNAs were detected in the pig blastocyst on days 13 and 16 of pregnancy , although the ovarian form was more abundant . Both forms of aromatase were much higher in day 13 than in day 16 pig blastocysts . The results provide definitive evidence that the preimplantation mouse blastocyst , as opposed to the pig blastocyst , has no ability to synthesize estrogen and no steroidogenic capacity . Maternal estrogen synthesis is essential for implantation of the mouse blastocyst . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . Steatohepatitis in laboratory opossums exhibiting a high lipemic response to dietary cholesterol and fat . Plasma VLDL and LDL cholesterol were markedly elevated ( > 40-fold ) in high-responding opossums , but moderately elevated ( 6-fold ) in low-responding opossums after they had consumed a high-cholesterol and high-fat diet for 24 wk . In both high- and low-responding opossums , plasma triglycerides were slightly elevated , threefold and twofold , respectively . Dietary challenge also induced fatty livers in high responders , but not in low responders . We studied the lipid composition , histopathological features , and gene expression patterns of the fatty livers . Free cholesterol ( 2-fold ) , esterified cholesterol ( 11-fold ) , and triglycerides ( 2-fold ) were higher in the livers of high responders than those in low responders , whereas free fatty acid levels were similar . The fatty livers of high responders showed extensive lobular disarray by histology . Inflammatory cells and ballooned hepatocytes were also present , as were perisinusoidal fibrosis and ductular proliferation . In contrast , liver histology was normal in low responders . Hepatic gene expression revealed differences associated with the development of steatohepatitis in high responders . The accumulation of hepatic cholesterol was concomitant with upregulation of the P04035 gene and downregulation of the Q02318 , Q9H221 , and P21439 genes . Genes involved in inflammation ( P01375 , P19838 , and P35354 ) and in oxidative stress ( P13498 and P14598 ) were upregulated . Upregulation of the growth factor genes ( PDGF and P01137 ) and collagen genes ( Col1A1 , Col3A1 , and Col4A1 ) was consistent with fibrosis . Some of the histological characteristics of the fatty livers of high-responding opossums imitate those in the livers of humans with nonalcoholic steatohepatitis . 27-hydroxycholesterol is an endogenous ligand for liver X receptor in cholesterol-loaded cells . The nuclear receptors liver X receptor alpha ( LXRalpha ) ( Q13133 ) and LXRbeta ( P55055 ) are important regulators of genes involved in lipid metabolism , including O95477 , P45844 , and sterol regulatory element-binding protein-1c ( SREBP-1c ) . Although it has been demonstrated that oxysterols are LXR ligands , little is known about the identity of the physiological activators of these receptors . Here we confirm earlier studies demonstrating a dose-dependent induction of O95477 and P45844 in human monocyte-derived macrophages by cholesterol loading . In addition , we show that formation of 27-hydroxycholesterol and cholestenoic acid , products of Q02318 action on cholesterol , is dependent on the dose of cholesterol used to load the cells . Other proposed LXR ligands , including 20(S)-hydroxycholesterol , 22(R)-hydroxycholesterol , and 24(S),25-epoxycholesterol , could not be detected under these conditions . A role for Q02318 in regulation of cholesterol-induced genes was demonstrated by the following findings . 1 ) Introduction of Q02318 into P29320 -293 cells conferred an induction of P45844 and SREBP-1c ; 2 ) upon cholesterol loading , Q02318 -expressing cells induce these genes to a greater extent than in control cells ; 3 ) in Q02318 -deficient human skin fibroblasts , the induction of O95477 in response to cholesterol loading was ablated ; and 4 ) in a coactivator association assay , 27-hydroxycholesterol functionally activated LXR . We conclude that 27-hydroxylation of cholesterol is an important pathway for LXR activation in response to cholesterol overload . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . Elevated retinol binding protein 4 induces apolipoprotein B production and associates with hypertriglyceridemia . CONTEXT AND OBJECTIVE : A high level of retinol binding protein 4 ( P02753 ) is reported to be associated with insulin resistance in humans . However , evidence from large-scale populations about the relationship between serum P02753 and metabolic phenotypes is scarce . In the present study , we aimed to evaluate serum P02753 distribution and its association with metabolic phenotypes among middle-aged and elderly Chinese . DESIGN AND PARTICIPANTS : Serum concentrations of P02753 in a cross-sectional sample of 2780 Chinese population aged 50-70 years old in Guangzhou were measured by ELISA . RESULTS : The mean of serum P02753 concentration was 28.04 μg/mL for male and 37.76 μg/mL for female ( P < .01 ) , respectively . Circulating P02753 was positively correlated with serum triglyceride and apolipoprotein B ( apoB ) concentrations . The odds ratio ( OR ) was substantially higher for hypertriglyceridemia ( OR , 3.26 ; 95 % confidence interval , 2.36-4.51 ) in the highest P02753 quartile compared with those in the lowest quartile after multiple adjustment for confounders . Furthermore , serum P02753 was significantly associated with fasting glucose , insulin levels , and homeostasis model assessment index-insulin resistance ( HOMA-IR ) . Moreover , we showed that P02753 enhanced microsomal triglyceride transfer protein ( P55157 ) expression and activity via up-regulation of protein disulfide isomerase ( P07237 ) , suppressed low-density lipoprotein receptor ( P01130 ) expression , and impaired insulin-signaling pathway , leading to inductions in apoB secretion both in vitro and in vivo . CONCLUSIONS : Elevated circulating P02753 concentrations were associated with higher risk of hypertriglyceridemia by inducing the secretion of triglyceride-rich apoB-containing lipoproteins . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . Mitochondrial cholesterol transport : a possible target in the management of hyperlipidemia . Q02318 ( Q02318 ) may defend cells against accumulation of excess cholesterol , making this enzyme a possible target in the management of hyperlipidemia . The study objective was to analyze cholesterol homeostatic responses to increases in Q02318 activity in HepG2 cells and primary human hepatocytes . Increasing Q02318 activity by increasing enzyme expression led to significant increases in bile acid synthesis with compensatory increases in P04035 ( HMGR ) activity/protein , P01130 ( P01130 ) mRNA , and P01130 -mediated cholesterol uptake . Under these conditions , only a small increase in cellular 27-hydroxycholesterol ( 27OH-Chol ) concentration was observed . No changes were detected in mature sterol regulatory element-binding proteins ( SREBP ) 1 or 2 . Increasing Q02318 activity by increasing mitochondrial cholesterol transport ( i.e. , substrate availability ) led to greater increases in bile acid synthesis with significant increases in cellular 27OH-Chol concentration . Mature SREBP 2 protein decreased significantly with compensatory decreases in HMGR protein . No change was detected in mature SREBP 1 protein . Despite increasing 27OH-Chol and lowering SREBP 2 protein concentrations , P01130 mRNA increased significantly , suggesting alternative mechanisms of P01130 transcriptional regulation . These findings suggest that regulation of liver mitochondrial cholesterol transport represents a potential therapeutic strategy in the treatment of hyperlipidemia and atherosclerosis . Interleukins 1α and 1β as regulators of steroidogenesis in human NCI-H295R adrenocortical cells . Inflammatory cytokines interleukin-1 ( IL-1 ) and tumor necrosis factor-α ( P01375 -α ) regulate the activity of the hypothalamo-pituitary-adrenal ( Q9Y251 ) axis at several levels . Although hypothalamic P06850 secretion may be the primary mechanism by which these cytokines activate the Q9Y251 axis , IL-1 expression is increased within the adrenal glands in models for systemic inflammation , and IL-1 may augment adrenal glucocorticoid production . Our aim was to investigate the direct effects of IL-1α and IL-1β on adrenal steroidogenesis and expression of three key steroidogenic genes in human adrenocortical cells using the NCI-H295R cell line as a model . mRNAs encoding receptors for IL-1 , P01375 -α , and leukemia inhibitory factor ( P15018 ) were detectable in the cell line ( Affymetrix microarray analysis ) . Both IL-1α and IL-1β increased cortisol , androstenedione , dehydroepiandrosterone and DB05804 production , and the accumulation of mRNAs for steroidogenic acute regulatory protein ( STAR ) , 17α-hydroxylase/17,20-lyase ( P05093 ) and 3β-hydroxysteroid dehydrogenase 2 ( P26439 ) in these cells ( P < 0.05 for all ) . Both ILs augmented P01375 -α- and P15018 -induced STAR and P05093 mRNA accumulation , and P01375 -α-induced cortisol production ( P < 0.05 for all ) . Both ILs also increased the apoptotic index of the cells ( P < 0.05 ) , which was efficiently neutralized by their specific antibodies . The IL-induced changes in the STAR , P26439 , and P05093 protein levels were not as evident as those in the respective mRNA levels . In conclusion , the combined effect of inflammatory cytokines at the adrenal level in acute or chronic inflammatory states could significantly stimulate glucocorticoid production , and thus explain the observed discrepancy between the cortisol and DB01285 concentrations sometimes seen in sepsis and chronic inflammatory states . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Identification of Reverb(alpha) as a novel ROR(alpha) target gene . The nuclear receptor superfamily comprises a large number of ligand-activated transcription factors that are involved in numerous biological processes such as cell proliferation , differentiation , and homeostasis . ROR(alpha) ( P35398 ) and Reverb(alpha) ( P20393 ) are two members of this family whose biological functions are largely unknown . In addition , no ligand has been yet identified for these two receptors ; therefore , they are referred as orphan receptors . Here , we show that ROR(alpha) and Reverb(alpha) are expressed with a similar tissue distribution and are both induced during the differentiation of rat Q9BTT4 myoblastic cells . Ectopic expression of ROR(alpha)1 in Q9BTT4 cells significantly induces Reverb(alpha) expression as demonstrated by Northern blot analysis . Using reverse transcription-PCR to analyze Reverb(alpha) gene expression from staggerer mice , we found that there was a significant reduction of Reverb(alpha) mRNA in the skeletal muscle comparing it with the wild-type mice , which suggests that ROR(alpha) is involved in the regulation of Reverb(alpha) gene expression . Transient transfection assays using the Reverb(alpha) promoter demonstrate that ROR(alpha) regulates the Reverb(alpha) gene at the transcriptional level . Furthermore , mutagenesis experiments indicate that ROR(alpha) regulates Reverb(alpha) transcription via a monomeric ROR response element located in the Reverb(alpha) gene promoter . Electrophoretic mobility shift assays show that ROR(alpha) binds strongly to this site in a specific-manner . Finally , overexpression of Q9Y3R0 / Q06418 -2 , but not Q15788 , potentiates ROR(alpha)-stimulated Reverb(alpha) promoter activity in transient transfection experiments . Together , our results identify Reverb(alpha) as a novel target gene for ROR(alpha) . Elevated expression of the genes encoding P01375 and thromboxane synthase in leucocytes from patients with systemic sclerosis . OBJECTIVE : To determine the expression of molecular markers of prostanoid/fatty acid signalling in leucocytes of patients with systemic sclerosis ( SSc ) . METHODS : Gene expression in patient leucocytes was analysed using real-time fluorescence reverse transcriptase polymerase chain reaction for tumour necrosis factor alpha ( P01375 ) , thromboxane synthase ( TXAS , CYP5A ) , prostacyclin synthase ( CYP8A ) , monocyte chemoattractant protein-1 ( P13500 ) , peroxisome proliferator-activated receptors ( Q07869 ) alpha , delta and gamma , low-density lipoprotein-associated lipoprotein lipase A(2) ( Q13093 ) , apolipoprotein E ( apoE ) and cholesterol 27-hydroxylase ( Q02318 ) . RESULTS : Both P01375 and TXAS showed an increase in mean expression in the diseased group ( 6.3-fold and 5.6-fold respectively , P < 0.0001 ) . These two markers , along with Q02318 , PPARgamma and apoE , provided predictive markers for the development of carotid artery disease within the SSc patient population . CONCLUSION : The elevated levels of P01375 and thromboxane seen in SSc patient sera are paralleled by increases in the expression of the appropriate genes in leucocytes . This method will allow us to screen for a large number of candidate markers of disease in order to increase our understanding of the processes underlying the pathology of SSc . Activation of c-Jun-N-terminal-kinase is crucial for the induction of a cell cycle arrest in human colon carcinoma cells caused by flurbiprofen enantiomers . The unselective cyclooxygenase ( P36551 ) inhibitor DB00712 and its-in terms of P36551 -inhibition- " inactive " enantiomer R-flurbiprofen have been previously found to inhibit tumor development and growth in various animal models . The underlying mechanisms are unknown . Here , we show that both R- and DB00712 reduce survival of three colon cancer cell lines , which differ in the expression of P35354 ( HCT-15 , no P35354 ; Caco-2 , inducible P35354 ; and HT-29 , constitutive P35354 ) . The IC50 for S- and R-flurbiprofen ranged from 250 to 450 microM . Both flurbiprofen enantiomers induced apoptosis in all three cell lines as indicated by DNA- and PARP-cleavage . In addition , R- and DB00712 caused a P55008 -cell cycle block . The latter was associated with an activation of c-Jun N-terminal kinase ( JNK ) , an increase of the DNA binding activity of the transcription factor AP-1 and down-regulation of cyclin D1 expression . Western blot analysis , as well as supershift experiments , revealed that the AP-1 activation was associated with a change of AP-1 composition toward an increase of JunB . The JNK inhibitor SP600125 antagonized R- and DB00712 -induced AP-1 DNA binding , suppression of cyclin D1 expression , and the P55008 -cell cycle block . However , JNK inhibition had no effect on flurbiprofen-induced apoptosis . Hence , the cell cycle arrest is obviously mediated , at least in part , through JNK-activation , whereas R- and DB00712 -induced apoptosis is largely independent of JNK . Although in vitro effects of R- and DB00712 were indistinguishable , only R-flurbiprofen inhibited HCT-15 tumor growth in nude mice , suggesting the involvement of additional in vivo targets , which are differently affected by R- and DB00712 . Expression of Th2-skewed pathology mediators in monocyte-derived type 2 of dendritic cells ( DC2 ) . The information conveyed from dendritic cells ( DCs ) to naïve P01730 (+) T cells has crucial influence on their differentiation toward effector T cells . In an effort to identify DC-derived molecules directly contributing to T cell differentiation , we searched for molecules distinctively expressed between two DC subtypes , which were differentiated from peripheral monocytes by cultivation with GM- P04141 ( for Q9NPG8 ) or P08700 ( for DC2 ) in the presence of P05112 and had the ability to induce naïve T cells to differentiate into Th1 or Th2 cells , respectively . As the first step to address this issue , we subtracted Q9NPG8 transcripts from those of DC2 and compiled the gene profile dominantly expressed in DC2 , whose products are known to reside in other than the nucleus . Intriguingly , many of them were molecules involved in Th2-skewed disease pathologies , such as P02751 , P38570 , Q14956 , Q03405 , P25089 , Q8NHJ6 , P05121 , P16050 , P24557 , P19878 , P10147 , P18510 , P09486 , and Q9NY15 , suggesting that DCs function not only as antigen presenting cells but also as producers of Th2 pathology specific milieus leading to disease deteriorations . We also found that expressions of Q02318 , O14495 , Q8WXG1 , and O15438 were up-regulated in DC2 , implying their significant function in Th2-deviated states . The identification of differentially expressed genes between DC subtypes provides new insights into their functions and our comparative gene expression profile will be highly useful for the identification of DC-derived key molecules for T cell differentiation .

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Efficacy , safety and tolerability of oral eletriptan in the acute treatment of migraine : results of a phase III , multicentre , placebo-controlled study across three attacks . The efficacy , safety and tolerability of the P28222 /D receptor agonist eletriptan ( 40 mg and 80 mg ) in acute treatment of migraine was evaluated in a multinational , randomized , double-blind , parallel-group , placebo-controlled , three-attack study treating 1153 patients . In the initial attack , significantly more eletriptan patients reported headache relief and complete pain relief at 2 h vs. placebo ( 40 mg 62 % and 32 % , 80 mg 65 % and 34 % , placebo 19 % and 3 % ; P < 0.0001 ) . Headache relief occurred faster after eletriptan , with more patients at both doses reporting relief 30 min ( P < 0.01 ) and 1 h ( P < 0.0001 ) after treatment than after placebo . There was a significantly lower recurrence rate with eletriptan 80 mg compared with placebo ( P < 0.01 ) . Adverse events for all treatments were generally mild or moderate and self-limiting . DB00216 40 mg and eletriptan 80 mg both appear to be effective and well-tolerated acute migraine treatments . Statin Modulation of Human T-Cell Proliferation , IL-1β and Q16552 Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD3/28-stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 (+) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 production ( P < 0.01 ) . However , in response to anti-CD3/28 stimulation , simvastatin significantly upregulated IL-1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells . Intramuscular gene transfer of P80511 inhibits neointimal hyperplasia after balloon injury in the rat abdominal aorta . P80511 is a well-known neuropeptide that has various protective effects on cardiovascular system . Our previous studies have shown that P80511 inhibits vascular smooth muscle cell ( VSMC ) proliferation in vitro . The present study aimed to explore the role of the P80511 in neointimal formation after balloon injury in the rat aortic wall and the underlying mechanism . Gene transfer of P80511 was performed with the use of intramuscular electroporation in a balloon-injured rat aorta model . Apoptosis in VSMCs was determined by electrophoresis assessment of DNA fragmentation and terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay . Overexpression of the P80511 gene significantly inhibited the neointimal formation after balloon injury compared with the mock transfer , as assessed by the intima-to-media ratio 14 days after balloon injury ( 29.2 +/- 3.7 % vs. 52.7 +/- 5.4 % ; n = 9-12 , P < 0.05 ) . In addition , P80511 gene expression increased the number of apoptotic cells in the neointima in vivo 14 days after balloon injury . Similarly , the addition of bioactive P80511 and the nitric oxide donor induced similar apoptosis in cultured VSMCs . The antagonist of the P80511 (1) receptor and inhibitors of DB02527 -PKA and nitric oxide blocked P80511 -mediated apoptosis . Furthermore , P80511 gene transfer increased inducible nitric oxide synthase and p53 but decreased P12004 and Bcl-2 protein levels in balloon-injured rat aorta . Our data demonstrated that P80511 potently inhibited neointimal thickening in the rat aorta , at least in part through its distinct effects on apoptosis and proliferation of VSMCs both in vivo and in vitro . Therefore , delivery of the P80511 gene may have therapeutic implications in limiting vascular restenosis . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor . Mouse pluripotent hematopoietic stem cells ( PHSC ) were fractionated based on size and density using counterflow centrifugal elutriation ( CCE ) . These heterogeneous PHSC populations were further enriched by subtraction of cells with lineage-specific markers ( Lin- ) followed by positive sorting for c-kit expression . The cells were characterized for their functional and biochemical properties . We defined a subpopulation of c-kit-positive cells that expressed high numbers of c-kit receptors ( c-kitBR ) . One hundred c-kitBR cells from either low- or higher-density fractions were sufficient to repopulate the lymphohematopoietic system in WBB6F1-W/Wv ( W/Wv ) recipients , whereas no PHSC were found in cells with low ( c-kitDULL ) or no ( c-kitNEG ) c-kit expression . Lin- c-kitBR cells were separated into RhoDULL and RhoBR subsets based on their ability to efflux rhodamine 123 ( Rho ) . The PHSC were concentrated in Lin- c-kitBR RhoDULL cells and the number of Lin- c-kitBR RhoBR cells correlated directly with the number of day 12 colony-forming unit-spleen ( CFU- P28222 ) in each fraction . We were not able to enrich further for PHSC using monoclonal antibodies to the cell-surface markers AA4.1 or P01730 , which have been used by others to isolate PHSC . The small , low-density Lin- c-kitBR subset contained PHSC and few CFU- P28222 . This enabled us to assay PHSC for expression of the flk-2 gene , which encodes a tyrosine kinase receptor present on fetal liver PHSC . Purified RNA from the low-density Lin- c-kitBR subset did not contain flk-2 mRNA . We suggest that AA4.1 , P01730 and flk-2 are expressed as stage-specific markers on PHSC in cell cycle . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Chk2-dependent phosphorylation of P18887 in the DNA damage response promotes base excision repair . The DNA damage response ( DDR ) has an essential function in maintaining genomic stability . Ataxia telangiectasia-mutated ( Q13315 ) -checkpoint kinase 2 ( Chk2 ) and Q13315 - and Rad3-related ( ATR ) -Chk1 , triggered , respectively , by DNA double-strand breaks and blocked replication forks , are two major DDRs processing structurally complicated DNA damage . In contrast , damage repaired by base excision repair ( BER ) is structurally simple , but whether , and how , the DDR is involved in repairing this damage is unclear . Here , we demonstrated that Q13315 -Chk2 was activated in the early response to oxidative and alkylation damage , known to be repaired by BER . Furthermore , Chk2 formed a complex with P18887 , the BER scaffold protein , and phosphorylated P18887 in vivo and in vitro at DB00156 (284) . A mutated P18887 lacking DB00156 (284) phosphorylation was linked to increased accumulation of unrepaired BER intermediate , reduced DNA repair capacity , and higher sensitivity to alkylation damage . In addition , a phosphorylation-mimic form of P18887 showed increased interaction with glycosylases , but not other BER proteins . Our results are consistent with the phosphorylation of P18887 by Q13315 -Chk2 facilitating recruitment of downstream BER proteins to the initial damage recognition/excision step to promote BER . Characterisation of the 5-HT receptor binding profile of eletriptan and kinetics of [3H]eletriptan binding at human P28222 and P28221 receptors . The affinity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] -1H-indole ) for a range of 5-HT receptors was compared to values obtained for other P28222 /1D receptor agonists known to be effective in the treatment of migraine . DB00216 , like sumatriptan , zolmitriptan , naratriptan and rizatriptan had highest affinity for the human P28222 , P28221 and putative 5-ht1f receptor . Kinetic studies comparing the binding of [3H]eletriptan and [3H]sumatriptan to the human recombinant P28222 and P28221 receptors expressed in HeLa cells revealed that both radioligands bound with high specificity ( > 90 % ) and reached equilibrium within 10-15 min . However , [3H]eletriptan had over 6-fold higher affinity than [3H]sumatriptan at the P28221 receptor ( K(D) ) : 0.92 and 6.58 nM , respectively ) and over 3-fold higher affinity than [3H]sumatriptan at the P28222 receptor ( K(D) : 3.14 and 11.07 nM , respectively ) . Association and dissociation rates for both radioligands could only be accurately determined at the P28221 receptor and then only at 4 degrees C . At this temperature , [3H]eletriptan had a significantly ( P < 0.05 ) faster association rate ( K(on) 0.249 min(-1) nM(-1) ) than [3H]sumatriptan ( K(on) 0.024 min(-1) nM(-1) ) and a significantly ( P < 0.05 ) slower off-rate ( K(off) 0.027 min(-1) compared to 0.037 min(-1) for [3H]sumatriptan ) . These data indicate that eletriptan is a potent ligand at the human P28222 , P28221 , and 5-ht1f receptors and are consistent with its potent vasoconstrictor activity and use as a drug for the acute treatment of migraine headache . P03372 -immunoreactive neurons contain calcitonin gene-related peptide , methionine-enkephalin or tyrosine hydroxylase in the female rat preoptic area . We have shown in our previous studies that estrogen treatment selectively influences calcitonin gene-related peptide ( P80511 ) - , methionine-enkephalin ( DB00134 -Enk ) - and tyrosine hydroxylase ( TH ) -immunoreactive ( IR ) intensities in the neurons of the periventricular preoptic nucleus ( Q9H237 ) and the medial preoptic area ( DB00603 ) of the female rat . In the present study , we examined whether estrogen receptor ( ER ) -IR neurons in the Q9H237 and DB00603 contain P80511 , DB00134 -Enk , or TH using a double-labeling immunohistochemical method and investigated changes in the number of double-labeling cells upon treatment with estrogen . Brain sections of ovariectomized rats and ovariectomized and estrogen-treated rat were stained using the avidin-biotin-peroxidase complex method followed by the peroxidase-anti-peroxidase method . The sections were first incubated with an anti-ER antibody in conjunction with nickel diaminobenzidine which produces a dark blue reaction product in the nucleus . Subsequently , P80511 , DB00134 -Enk or TH antisera were applied to these sections and the resulting brown diaminobenzidine reaction product in the cytoplasm was examined . Neurons that were double-labeled for ER and P80511 , DB00134 -Enk or TH were investigated in the Q9H237 and DB00603 . The number of doubly labeled ER/ P80511 - and ER/TH-IR neurons was large , whereas the number of ER/ DB00134 -Enk-IR neurons was small . These results suggest that ER in the Q9H237 and DB00603 may be more closely related to the mechanism of changes in P80511 - and TH-IR intensities upon estrogen treatment than that in DB00134 -Enk-IR intensity . Increased serum levels of P05109 /A9 and P80511 are associated with cardiovascular disease in patients with inactive systemic lupus erythematosus . OBJECTIVES : Patients with SLE have an increased morbidity and mortality from cardiovascular disease ( CVD ) . The reason for this is not entirely understood , but is believed to be partly related to the long-lasting inflammatory process seen in SLE . The aim of the present study was to investigate whether there is an association between CVD and serum levels of the proinflammatory proteins P05109 /A9 and P80511 in SLE . METHODS : Serum levels of P05109 /A9 and P80511 were measured with ELISA in 237 SLE patients with clinically inactive disease and without infections , as well as in 100 healthy individuals . Cardiovascular manifestations were defined according to the SLICC/ P10323 Damage Index ( SLICC/ P10323 -DI ) . RESULTS : Serum levels of P05109 /A9 were elevated in our inactive SLE patients as compared with healthy individuals ( P < 0.0001 ) , which was not seen for P80511 ( P = 0.12 ) . SLE patients with a history of CVD had increased serum levels of both P05109 /A9 and P80511 compared with patients with no CVD or venous thromboembolism ( P = 0.003 and P = 0.006 , respectively ) . The presence of organ damage according to SLICC/ P10323 -DI was associated with an increase in both P05109 /A9 and P80511 serum levels ( P = 0.001 and P = 0.006 , respectively ) . CONCLUSION : Elevated serum levels of P05109 /A9 and P80511 may be used as an indicator of severe disease and CVD in SLE , suggesting that SLE patients with elevated serum P05109 /A9 and P80511 concentrations may benefit from more intense cardiovascular primary preventive strategies and possibly also from more intense and early immunosuppressive treatment . Characterisation of the contractile activity of eletriptan at the canine vascular P28222 receptor . The functional activity of eletriptan ( ( R ) -3-(1-methyl-2-pyrrolidinylmethyl)-5- [ 2- ( phenylsulphonyl ) ethyl ] - 1 H-indole ) at the contractile serotonin ( 5-hydroxytryptamine ; 5-HT ) ' 1B-like ' receptor in dog isolated saphenous vein and basilar artery was investigated . DB00216 , like 5-HT and sumatriptan potently contracted saphenous vein ( pEC50 : 6.3 , 6.9 and 6.1 , respectively ) and basilar artery ( pEC50 7.2 , 7.5 and 6.8 , respectively ) . The maximum responses evoked by eletriptan was , unlike sumatriptan , significantly lower than that to 5-HT ( intrinsic activity saphenous vein : eletriptan 0.57 , 5-HT 1.0 , sumatriptan 0.85 ; basilar artery : eletriptan 0.77 , 5-HT 0.98 , sumatriptan 0.89 ) . Contractions evoked by eletriptan were antagonised by the P28222 /1D receptor antagonist GR125743 ( N- [ 4-methoxy-3- ( 4-methyl piperazin-1-yl ) phenyl ] -3-methyl-4-(4-pyridyl)benzamide ) with pA2 values of 9.1 in saphenous vein and 9.4 in basilar artery . Affinity estimates ( pKA ) for 5-HT and sumatriptan determined from receptor alkylation studies in saphenous vein were 6.6 and 6.3 , respectively , compared to the apparent equilibrium dissociation constant ( pKp ) for eletriptan of 6.8 . The rank order of relative intrinsic efficacies ( epsilon ) was 5-HT > sumatriptan > eletriptan . Thus , eletriptan required greater receptor occupancy ( 4.4-fold ) to evoke an equivalent contraction to 5-HT and sumatriptan in dog isolated saphenous vein . These data demonstrate that eletriptan is a potent partial agonist at the canine vascular P28222 receptor . Prolonged treatment with bicalutamide induces androgen receptor overexpression and androgen hypersensitivity . BACKGROUND : Various hormone refractory prostate cancer cell models have been established with androgen depletion and have helped to clarify the mechanism for the transition into androgen-depletion independent status . However , the mechanism of bicalutamide resistance remains unclear because few cell models have been generated . METHODS : We generated a bicalutamide-resistant subline , LNCaP- O43633 , from LNCaP after prolonged treatment with bicalutamide . Androgen and/or bicalutamide responsiveness for proliferation and prostate-specific antigen ( PSA ) secretion were examined in vitro and in vivo . DB00624 and dihydrotestosterone ( DB02901 ) levels in xenografted tumors were analyzed by liquid chromatography-tandem mass spectrometry . P10275 ( AR ) gene mutation and amplification and AR and pAR(210) expression were determined . RESULTS : LNCaP- O43633 did not grow in an androgen-depleted medium and proliferation was stimulated in a tenfold lower concentration of androgen than that of LNCaP . LNCaP- O43633 grew in castrated male mice , and the DB02901 level in grafted LNCaP- O43633 tumors was 7.7-fold lower than in LNCaP tumors . DB01128 stimulated LNCaP- O43633 proliferation and PSA secretion in vitro and the antitumor activity of bicalutamide against LNCaP- O43633 was weaker than that of LNCaP in vivo . Additional AR mutation and AR gene amplification were not detected in LNCaP- O43633 , but AR and pAR(210) expression and PSA secretion in LNCaP- O43633 were higher than in LNCaP . CONCLUSIONS : DB01128 -resistant LNCaP- O43633 exhibited AR overexpression and hypersensitivity to low levels of androgen . Our data suggests that AR overexpression is a significant mechanism of bicalutamide resistance similar to resistance from chronic androgen depletion . In addition , pAR(210) overexpression could be a potential mechanism for hypersensitivity to low androgen in LNCaP- O43633 . The effects of aging and chronic fluoxetine treatment on circadian rhythms and suprachiasmatic nucleus expression of neuropeptide genes and P28222 receptors . Age-related changes in circadian rhythms , including attenuation of photic phase shifts , are associated with changes in the central pacemaker in the suprachiasmatic nucleus ( SCN ) . Aging decreases expression of mRNA for vasoactive intestinal peptide ( P01282 ) , a key neuropeptide for rhythm generation and photic phase shifts , and increases expression of serotonin transporters and 5-HT(1B) receptors , whose activation inhibits these phase shifts . Here we describe studies in hamsters showing that aging decreases SCN expression of mRNA for gastrin-releasing peptide , which also modulates photic phase resetting . Because serotonin innervation trophically supports SCN P01282 mRNA expression , and serotonin transporters decrease extracellular serotonin , we predicted that chronic administration of the serotonin-selective reuptake inhibitor , fluoxetine , would attenuate the age-related changes in SCN P01282 mRNA expression and 5-HT(1B) receptors . In situ hybridization studies showed that fluoxetine treatment does not alter SCN P01282 mRNA expression , in either age group , at zeitgeber time (ZT)6 or 13 ( ZT12 corresponds to lights off ) . However , receptor autoradiographic studies showed that fluoxetine prevents the age-related increase in SCN 5-HT(1B) receptors at ZT6 , and decreases SCN 5-HT(1B) receptors in both ages at ZT13 . Therefore , aging effects on SCN P01282 mRNA and SCN 5-HT(1B) receptors are differentially regulated ; the age-related increase in serotonin transporter sites mediates the latter but not the former . The studies also showed that aging and chronic fluoxetine treatment decrease total daily wheel running without altering the phase of the circadian wheel running rhythm , in contrast to previous reports of phase resetting by acute fluoxetine treatment . Loss of both phospholipid and triglyceride transfer activities of microsomal triglyceride transfer protein in abetalipoproteinemia . Mutations in microsomal triglyceride transfer protein ( P55157 ) cause abetalipoproteinemia ( P00519 ) , characterized by the absence of plasma apoB-containing lipoproteins . In this study , we characterized the effects of various P55157 missense mutations found in P00519 patients with respect to their expression , subcellular location , and interaction with protein disulfide isomerase ( P07237 ) . In addition , we characterized functional properties by analyzing phospholipid and triglyceride transfer activities and studied their ability to support apoB secretion . All the mutants colocalized with calnexin and interacted with P07237 . We found that R540H and N780Y , known to be deficient in triglyceride transfer activity , also lacked phospholipid transfer activity . Novel mutants S590I and G746E did not transfer triglycerides and phospholipids and did not assist in apoB secretion . In contrast , D384A displayed both triglyceride and phospholipid transfer activities and supported apoB secretion . These studies point out that P00519 is associated with the absence of both triglyceride and phospholipid transfer activities in P55157 . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . Porcine carotid vascular effects of eletriptan ( UK-116,044 ) : a new P28222 /1D receptor agonist with anti-migraine activity . It has been suggested that opening of cephalic arteriovenous anastomoses may be involved in the headache phase of migraine . Indeed , a number of acutely acting anti-migraine drugs , including the ergot alkaloids and sumatriptan , constrict porcine carotid arteriovenous anastomoses . In this study , using pentobarbital anaesthetised pigs , we investigated the effects of eletriptan , a close structural analogue of sumatriptan , on the distribution of common carotid artery blood flow into arteriovenous anastomotic and nutrient ( capillary ) fractions . DB00216 ( 10 , 30 , 100 , 300 and 1000 microg kg(-1) , i.v. ) decreased the total carotid blood flow , exclusively by decreasing cephalic arteriovenous anastomotic blood flow ; nutrient blood flow , particularly to the ear , skin and fat , was significantly increased . The doses of eletriptan needed to reduce arteriovenous anastomotic blood flow and conductance by 50 % ( ED50 ) were , respectively , 117+/-21 microg kg(-1) ( 251+/-45 nmol kg(-1) ) and 184+/-42 microg kg(-1) ( 396+/-91 nmol kg(-1) ) ; the highest dose caused reductions of 84+/-3 % and 77+/-4 % , respectively . The eletriptan-induced changes in carotid haemodynamics were clearly attenuated by pretreating the pigs with the selective P28222 /1D receptor antagonist GR127935 ( 0.5 mg kg(-1) ) . On the basis of these results , we conclude that ( 1 ) the eletriptan-induced constriction of cephalic arteriovenous anastomoses as well as the arteriolar dilatation in head tissues is predominantly mediated by P28222 /1D receptors , and ( 2 ) eletriptan should be effective in aborting migraine headache . Clinical studies have already demonstrated its therapeutic action in migraine patients . The Mek1 phosphorylation cascade plays a role in meiotic recombination of Schizosaccharomyces pombe . Mek1 is a Chk2/Rad53/Cds1-related protein kinase that is required for proper meiotic progression of Schizosaccharomyces pombe . However , the molecular mechanisms of Mek1 regulation and Mek1 phosphorylation targets are unclear . Here , we report that Mek1 is phosphorylated at serine-12 ( P28222 ) , Q92748 and threonine-15 ( T15 ) by Rad3 ( ATR ) and/or Tel1 ( Q13315 ) kinases that are activated by meiotic programmed double-strand breaks ( DSBs ) . Mutations of these sites by alanine replacement caused abnormal meiotic progression and recombination rates . Phosphorylation of these sites triggers autophosphorylation of Mek1 ; indeed , alanine replacement mutations of Mek1-T318 and -T322 residues in the activation loop of Mek1 reduced Mek1 kinase activity and meiotic recombination rates . Substrates of Mek1 include Mus81-T275 , Rdh54- Q8NHM4 and Rdh54-T673 . Mus81-T275 is known to regulate the Mus81 function in DNA cleavage , whereas Rdh54-T6A/T673A mutant cells showed abnormal meiotic recombination . Taken together , we conclude that the phosphorylation of Mek1 by Rad3 or Tel1 , Mek1 autophosphorylation and Mus81 or Rdh54 phosphorylation by Mek1 regulate meiotic progression in S. pombe . Antidepressant effects on serotonin 1A/1B receptors in the rat brain using a gene x environment model . A gene-environment ( GxE ) interaction is implicated in both the pathophysiology and treatment of major depressive disorder ( MDD ) . This study modeled the effects of genetic vulnerability by using the Flinders sensitive line ( FSL ) , a rat model of depression and its control counterpart-the Flinders resistant line ( FRL ) . The effects of environmental vulnerability ( e.g. , early-life stress ) were modeled by using maternal separation . Rats ( n=105 ) were drawn from four groups reflecting experimental crossing of strain ( FSL vs. FRL ) and early-life stress ( high vs. low ) to assess the effects of two antidepressants ( escitalopram or nortriptyline ) compared to vehicle . Quantitative in vitro autoradiography was performed using [(125)I]MPPI ( P08908 ) and [(125)I]CYP ( P28222 ) in prefrontal cortex ( P27918 ) and hippocampus . Stringent , Bonferroni-corrected statistical analyses showed significant strain-by-rearing-by-treatment ( three-way ) interactions in P27918 P08908 and hippocampal P28222 receptors . Either vulnerability reduced serotonergic binding ; no additive effects were associated with the two vulnerabilities . Both antidepressants increased hippocampal P28222 receptor binding ; however , only nortriptyline selectively increased P27918 P08908 receptor binding . Taken together , our findings demonstrate that antidepressant effects on the serotonergic system are shaped by a GxE interaction that depends on antidepressant class and brain region . Increased expression of receptor for advanced glycation end products by synovial tissue macrophages in rheumatoid arthritis . OBJECTIVE : The accumulation of advanced glycation end products ( AGEs ) , P80511 , and high mobility group box chromosomal protein 1 has been associated with joint inflammation in rheumatoid arthritis ( RA ) . This study was undertaken to determine the induction of the receptor for these proteins , termed receptor for AGEs ( RAGE ) , in synovial tissue ( ST ) macrophages from RA patients . METHODS : RAGE and P34810 expression in ST were determined by 2-color immunofluorescence labeling . Cell surface and messenger RNA ( mRNA ) expression of RAGE were examined by flow cytometry and reverse transcriptase-polymerase chain reaction ( PCR ) or real-time PCR , respectively . RESULTS : P34810 + lining macrophages , like the vasculature , expressed high levels of RAGE in inflamed ST from RA patients . RAGE mRNA expression was significantly higher in RA ST than in ST from patients with osteoarthritis . RAGE mRNA levels were significantly higher in ST macrophages and normal endothelial cells than in ST P01730 + T cells and synovial fibroblasts stimulated with tumor necrosis factor alpha and interleukin-1beta ( IL-1beta ) . Cell surface RAGE was highly induced on normal monocytes after a 24-hour incubation with a 20 % concentration of RA ST cell culture supernatants . RAGE mRNA expression in adherent monocytes was augmented by various cytokines , most potently by IL-1beta . CONCLUSION : These results indicate that RAGE overexpression in lining macrophages may be induced , at least in part , by cytokines such as IL-1 , leading to the amplification of inflammatory responses mediated by RAGE ligands that are abundant in RA joints . Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients . Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 ( CYP ) enzymes . In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A ( Q13216 ) , a substrate of P08684 . The effects of azithromycin on Q13216 disposition kinetics were evaluated in eight stable renal transplant patients . Patients had been stabilized on individualized doses of Q13216 which remained unchanged throughout the study . DB00207 was administered for 3 days . Baseline measurements of Q13216 disposition kinetics were taken prior to azithromycin treatment ( study day 2 ) and after 3 days ( study day 5 ) of azithromycin treatment ( 500mg/day , orally ) . The key parameters of interest were the area under the Q13216 blood concentration versus time curve ( AUC ) measured for 24h after the morning dose of Q13216 on both days 2 and 5 , and the C(max) values of Q13216 . The geometric mean ratios ( GMRs ) of those parameters ( day 5/day 2 ) and their 90 % confidence intervals ( 90 % CI ) were 107 ( 98,116 ) and 119 ( 104,136 ) , respectively . The 7 % increase in exposure level and 19 % increase in peak plasma concentration are not likely to be clinically significant . It is concluded that azithromycin ( 500mg/dayx3 days ) does not alter the disposition kinetics of Q13216 in a clinically significant way , and that Q13216 dosage adjustments are not warranted in renal transplant patients taking these two drugs together . DB08827 : A novel agent for the treatment of homozygous familial hypercholesterolemia . PURPOSE : The pharmacology , pharmacokinetics , and clinical efficacy and safety of lomitapide in the management of homozygous familial hypercholesterolemia ( HoFH ) are reviewed . SUMMARY : DB08827 ( Juxtapid , Aegerion Pharmaceuticals ) is an oral microsomal triglyceride transfer protein ( P55157 ) inhibitor indicated for the treatment of patients with HoFH , a rare form of hypercholesterolemia that can lead to premature atherosclerotic disease . In clinical trials , the use of lomitapide alone or in combination with other lipid-lowering modalities reduced plasma concentrations of low-density lipoprotein cholesterol ( LDL-C ) by a mean of more than 50 % . DB08827 is associated with significant gastrointestinal adverse effects and increases in hepatic fat levels . DB08827 undergoes hepatic metabolism via cytochrome P-450 ( CYP ) isoenzyme 3A4 and interacts with P08684 substrates including atorvastatin and simvastatin ; dose adjustment is recommended when lomitapide is used concurrently with these agents . In patients receiving concomitant warfarin , the International Normalized Ratio ( INR ) should be closely monitored , as lomitapide use may increase INR values . The recommended initial dosage of lomitapide is 5 mg once daily , with subsequent upward dose adjustment at specified intervals according to tolerability . DB08827 is contraindicated in patients with moderate-to-severe liver disease , patients with sustained abnormal liver function tests , patients taking strong or moderate P08684 inhibitors , and pregnant patients . CONCLUSION : DB08827 is an oral P55157 inhibitor approved for the treatment of HoFH . This agent appears to be a realistic option for patients with HoFH who are unable to attain their LDL-C goal or can not tolerate statin therapy . P10275 and calcitonin gene-related peptide in neurons of the genitofemoral nerve during testicular descent induced with human chorionic gonadotropin . BACKGROUND : Low levels of circulating testosterone during testis descent cause cryptorchidism in humans and rats . Treatment with human chorionic gonadotrophin ( hCG ) induces testis descent by stimulating production of testosterone ( T ) . Neurons of genitofemoral nerve ( GFN ) , which innervate testicular gubernaculum , may play a role in testis descent . METHODS : In the current study , putative correlations were made between T and GFN motor and sensory neuron activity during inguinoscrotal testis descent . Cryptorchidism was provoked in prepuberal rats with estradiol . Rats with testicular descent induced with hCG and cryptorchid controls were used . Cells of spinal cord and dorsal root ganglia were labeled by retrograde staining with fast-blue . Expression of androgen receptor ( AR ) and calcitonin gene-related peptide ( P80511 ) were detected with indirect immunofluorescence . RESULTS : Neurons labeled with fast-blue were found in the center of motor horn and dorsal root ganglia at levels Q9NUQ9 and Q401N2 . While number of motor neurons expressing AR was significantly higher in the group treated with hCG , number expressing P80511 was higher in controls . In dorsal root ganglion , number of cells immunostained with P80511 antibody was similar in both groups but AR was not detected . CONCLUSIONS : Present results support the hypothesis that motor nucleus of the GFN is a direct target of testosterone and that regulation of P80511 in sensory nucleus may be involved in testicular descent . Effects of eletriptan on the peptidergic innervation of the cerebral dura mater and trigeminal ganglion , and on the expression of c-fos and c-jun in the trigeminal complex of the rat in an experimental migraine model . Nociceptive axons and terminals in the supratentorial cerebral dura mater display an intense calcitonin gene-related peptide ( P80511 ) immunoreactivity . In an experimental migraine model , it has been shown that electrical stimulation of the rat trigeminal ganglion induced an increase in the lengths of P80511 -immunoreactive axons , increased size and number of pleomorphic axonal varicosities in the dura mater , and an increased number of c-jun and c-fos protein-expressing nerve cells in the trigeminal complex . We demonstrate the effect of the highly specific and moderately lipophilic serotonin agonist eletriptan ( Pfizer ) which prevents the effects of electrical stimulation in the dura mater . DB00216 also affected the caudal trigeminal complex ; it markedly reduced the numbers of the oncoprotein-expressing cells , mainly after stimulation and to some extent also in nonstimulated animals . DB00216 also affected expression of P80511 in perikarya of trigeminal ganglion cells , insofar as the number of small nerve cells exhibiting a compact P80511 immunoreaction was decreased to one quarter of the original value . In all these respects , eletriptan acted in a similar way to sumatriptan , with the notable exception that eletriptan also blocked the stimulation-induced effects in the nucleus caudalis trigemini and the upper cervical spinal cord ( trigeminal complex ) , whereas sumatriptan did not . It is concluded that eletriptan , acting on perikarya and both the peripheral and the central axon terminals of primary sensory neurons , exerts its antimigraine effect by an agonist action on P28222 /1D receptors throughout the entire trigeminal system , probably by passing the blood-brain-barrier because of its lipophilic character . DB00216 Pfizer . Pfizer has developed and launched eletriptan , a P28222 /1D agonist , for the potential treatment of migraine with and without aura . DB00216 has 6-fold greater affinity for the P28221 receptor than sumatriptan , and a 3-fold greater affinity for the P28222 receptor [ 249570 ] . DB00216 pharmacology has also been evaluated in vitro in comparison with zolmitriptan ( AstraZeneca plc ) and naratriptan ( GlaxoSmithKline plc ) [ 290116 ] . Identification of cellular pathways of " type 1 , " Th17 T cells , and P01375 - and inducible nitric oxide synthase-producing dendritic cells in autoimmune inflammation through pharmacogenomic study of cyclosporine A in psoriasis . Therapeutic modulation of psoriasis with targeted immunosuppressive agents defines inflammatory genes associated with disease activity and may be extrapolated to a wide range of autoimmune diseases . DB00091 A ( Q13216 ) is considered a " gold standard " therapy for moderate-to-severe psoriasis . We conducted a clinical trial with Q13216 and analyzed the treatment outcome in blood and skin of 11 responding patients . In the skin , as expected , Q13216 modulated genes from activated T cells and the " type 1 " pathway ( p40 , P01579 , and P35610 -1-regulated genes ) . However , Q13216 also modulated genes from the newly described Th17 pathway ( Q16552 , Q9GZX6 , and downstream genes P80511 , DEFB-2 , IL-1beta , SEPRINB3 , P80188 , and P78556 ) . Q13216 also affected dendritic cells , reducing P01375 and inducible NO synthase ( products of inflammatory P01375 - and inducible NO synthase-producing dendritic cells ) , Q01151 , and Q9NPF7 . We detected 220 early response genes ( day 14 posttreatment ) that were down-regulated by Q13216 . We classified > 95 % into proinflammatory or skin resident cells . More myeloid-derived than activated T cell genes were modulated by Q13216 ( 54 myeloid genes compared with 11 lymphocyte genes ) , supporting the hypothesis that myeloid derived genes contribute to pathogenic inflammation in psoriasis . In circulating mononuclear leukocytes , in stark contrast , no inflammatory gene activity was detected . Thus , we have constructed a genomic signature of successful treatment of psoriasis which may serve as a reference to guide development of other new therapies . In addition , these data also identify new gene targets for therapeutic modulation and may be applied to wide range of autoimmune diseases . P40189 -linked signal transduction promotes the differentiation and maturation of dendritic cells . In order to explore the role of P40189 -linked signal transduction in the differentiation and maturation of dendritic cells ( DC ) , the mAb , B- P28222 , an agonist of P40189 , was used for the activation of P40189 on DC . The effects of cytokines and of anti- P40189 mAb on the proliferation of DC , and their expression of IL-12 and P33681 ( P33681 -1 ) by DC were evaluated . DC differentiating from peripheral blood mononuclear cells did not express the P05231 receptor alpha chain , but expressed P40189 . Anti- P40189 mAb promoted the proliferation of DC , induced by P05112 and granulocyte macrophage colony stimulating factor ( GM- P04141 ) , by up-regulating the GM- P04141 receptor on DC . DC induced by P40189 mAb and cytokines expressed DC-derived CC chemokine , as measured by RT-PCR . Induced DC also stimulated strong proliferation of autologous T cells in mixed lymphocyte reaction since an up-regulated expression of IL-12 and P33681 ( P33681 -1 ) was observed in DC activated by anti- P40189 mAb . Thus , P40189 signal transduction is important for the differentiation and maturation of DC . Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 ) of the hypothalamus . Animals were treated during early puberty , from P01160 23 to P01160 30 , with EE and Q03001 given orally every day . They were then sacrificed and perfused on P01160 37 or P01160 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 . EE and Q03001 increased ER-labelled neurons in the ARC and DB00603 . At P01160 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 increased ER-labelled neurons in the DB00603 in females . EE increased testosterone in males at P01160 37 and estradiol in females at P01160 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats . Serotonergic modulation of the acoustic startle response in rats during preweaning development . The involvement of serotonin ( 5-HT ) in modulating the acoustic startle response ( ASR ) is well established in adult rats , but 5-HT involvement during the preweaning period , when 5-HT neurons undergo extensive development , has not previously been described . Three 5-HT receptor subtypes are reported to modulate the ASR in adult rats : P08908 and 5-HT2 receptor agonists facilitate the ASR , whereas P28222 agonists decrease the response . In the present study , the effects of 5-HT agonists and generalized 5-HT depletion on the ASR were studied in preweanling animals , using independent groups of Long-Evans rats tested on postnatal day ( P01160 ) 13 , 17 and 21 . 8-Hydroxy-2-(di-n-propylamino) tetralin ( 8OHDPAT , 62-1000 micrograms/kg ) , a P08908 receptor agonist , and 5-methoxy-N,N-dimethyl tryptamine ( MeODMT , 2-4 mg/kg ) , a nonselective 5-HT agonist , had no effect on P01160 13 and then increased the ASR on P01160 17 and 21 . The 5-HT2 receptor antagonists cyproheptadine ( 5 mg/kg ) and ketanserin ( 5 mg/kg ) blocked the effect of MeODMT at both ages , providing some evidence that MeODMT increased the ASR through 5-HT2 receptors . 1-(m-Chlorophenyl) piperazine ( mCPP , 1-5 mg/kg ) , a P28222 agonist , had no effect on ASR amplitude on P01160 13 or 17 and then produced a dose-related decrease in the response on P01160 21 . Generalized depletion of 5-HT by 80-90 % in whole-brain and spinal cord , using p-chlorophenylalanine ( PCPA , 300 mg/kg 24 hr prior to testing ) , did not alter ASR amplitude at any age. ( ABSTRACT TRUNCATED AT 250 WORDS ) MnSOD drives neuroendocrine differentiation , androgen independence , and cell survival in prostate cancer cells . An increase in neuroendocrine ( NE ) cell number has been associated with progression of prostate tumor , one of the most frequent cancers among Western males . We previously reported that mitochondrial manganese superoxide dismutase ( MnSOD ) increases during the NE differentiation process . The goal of this study was to find whether MnSOD up-regulation is enough to induce NE differentiation . Several human prostate cancer LNCaP cell clones stably overexpressing MnSOD were characterized and two were selected ( MnSOD-S4 and MnSOD- P28222 ) . MnSOD overexpression induces NE morphological features as well as coexpression of the NE marker synaptophysin . Both MnSOD clones exhibit lower superoxide levels and higher H(2)O(2) levels . MnSOD-overexpressing cells show higher proliferation rates in complete medium , but in steroid-free medium MnSOD- P28222 cells are still capable of proliferation . MnSOD up-regulation decreases androgen receptor and prevents its nuclear translocation . MnSOD also induces up-regulation of Bcl-2 and prevents docetaxel- , etoposide- , or P01375 -induced cell death . Finally , MnSOD-overexpressing cells enhance growth of androgen-independent PC-3 cells but reduce growth of androgen-dependent cells . These results indicate that redox modulation caused by MnSOD overexpression explains most NE-like features , including morphological changes , NE marker expression , androgen independence , inhibition of apoptosis , and enhancement of cell growth . Many of these events can be associated with the androgen dependent-independent transition during prostate cancer progression . DB00819 inhibits osmotic water permeability by interaction with aquaporin-1 . DB09145 channel proteins , known as aquaporins , are transmembrane proteins that mediate osmotic water permeability . In a previous study , we found that acetazolamide could inhibit osmotic water transportation across Xenopus oocytes by blocking the function of aquaporin-1 ( P29972 ) . The purpose of the current study was to confirm the effect of acetazolamide on water osmotic permeability using the human embryonic kidney 293 ( HEK293 ) cells transfected with pEGFP/ P29972 and to investigate the interaction between acetazolamide and P29972 . The fluorescence intensity of HEK293 cells transfected with pEGFP/ P29972 , which corresponds to the cell volume when the cells swell in a hyposmotic solution , was recorded under confocal laser fluorescence microscopy . The osmotic water permeability was assessed by the change in the ratio of cell fluorescence to certain cell area . DB00819 , at concentrations of 1 and 10muM , inhibited the osmotic water permeability in HEK293 cells transfected with pEGFP/ P29972 . The direct binding between acetazolamide and P29972 was detected by surface plasmon resonance . P29972 was prepared from rat red blood cells and immobilized on a CM5 chip . The binding assay showed that acetazolamide could directly interact with P29972 . This study demonstrated that acetazolamide inhibited osmotic water permeability through interaction with P29972 . DB00819 inhibits stimulated feline liver and gallbladder bicarbonate secretion . Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation . P00918 ( CA II ) , that is inhibited by acetazolamide , plays a role in regulation of the acid-base balance in many tissues . This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide ( P01282 ) -stimulated gallbladder mucosal bicarbonate and acid secretion . Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air . In 20 experiments P01282 ( 10 microg kg(-1) h(-1) ) and in 10 experiments secretin ( 4 microg kg(-1) h(-1) ) were infused continuously intravenous ( i.v. ) . Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport . During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen . Intravenous infusion of vasoactive intestinal peptide ( P01282 ) and secretin caused a secretion of bicarbonate from the gallbladder mucosa ( P < 0.01 ) . This secretion was reduced by intraluminal ( i.l. ) acetazolamide ( P < 0.01 ) . Bile flow was enhanced by infusion of P01282 and secretin ( P < 0.01 ) but this stimulated outflow was not affected by i.v. acetazolamide . The presence of CA II in the gallbladder was demonstrated by immunoblotting . Biliary CA activity has an important function in the regulation of P01282 - and secretin-stimulated bicarbonate secretion across the gallbladder mucosa . Neurochemical properties of aquaporin 1-expressing sensory neurons from the ovine trigeminal ganglion . Aims of the present study were to investigate the distribution and morphology of aquaporin 1-immunoreactive ( P29972 -IR ) neurons in the sensory ganglia of the sheep . Double immunohistochemical staining was applied to figure out whether DB05875 ( SP ) , calcitonin gene-related peptide ( P80511 ) and galanin are present in P29972 -bearing primary afferent neurons . The expression of P29972 was present only in trigeminal ganglion , whereas in nodose ganglion , jugular ganglion as well as C(1) -C(7) dorsal root ganglia no presence of P29972 was found . In trigeminal ganglion , 15.4 ± 2.3 % of Hu C/D-IR neurons ( pan-neuronal marker ) showed the presence of P29972 . The vast majority of P29972 -IR trigeminal sensory neurons ( approximately 69.6 ± 3.3 % , n = 5 ) were classified as middle in size , 28.6 ± 3.0 % of P29972 -IR neurons were small and only 1.8 ± 0.6 % of P29972 -positive neurons were large in size . Amongst the population of P29972 -IR trigeminal neurons as many as 58.5 ± 3.9 % were immunopositive to SP , 30.7 ± 2.3 % showed the presence of P80511 and 10.9 ± 0.2 % coexpressed galanin . In trigeminal ganglion , SP-IR as well as P80511 -IR ( but not galanin-IR ) nerve fibres were found in close neighbourhood of P29972 -IR neurons . It is concluded that P29972 is present in certain neuronal subsets of the ovine trigeminal ganglion ; however , the exact role of this water channel has to be elucidated . [ Signal transduction inhibitor -- STI571 -- a new treatment for chronic myeloid leukemia ( CML ) , which opens a new targeted approach to cancer therapy ] . Chronic myeloid leukemia ( CML ) , in most of the cases , is the molecular consequence of the t(9,22) translocation , resulting in the Philadelphia ( Ph ) chromosome and the creation of the fusion gene P11274 - P00519 . The fusion gene is translated to the protooncogen P11274 - P00519 , a constitutively activated tyrosine kinase that is linked to the malignant transformation . Thus , this tyrosine kinase became an attractive target for drug design . The development of the novel investigational drug DB00619 is based on its potent and selective ability to inhibit this fusion tyrosine kinase . In preclinical studies , DB00619 selectively inhibited the growth of CML cells that carry the Ph chromosome . In this review we discuss the drug development and design , its mechanism of action , the preclinical studies and the results of phase I and II clinical trials . Array-comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas . Oral squamous cell carcinoma ( OSCC ) is a common worldwide malignancy . However , it is unclear what , if any , genomic alterations occur as the disease progresses to invasive and metastatic OSCC . This study used genomewide array-CGH in microdissected specimens to map genetic alterations found in primary OSCC and neck lymph node metastases . We used array-based comparative genomic hybridization ( array-CGH ) to screen genomewide alterations in eight pairs of microdissected tissue samples from primary and metastatic OSCC . In addition , 25 primary and metastatic OSCC tissue pairs were examined with immunohistochemistry for protein expression of the most frequently altered genes . The highest frequencies of gains were detected in P12524 , Q04864 , TERC , P42336 , P10242 , P08183 , P01112 , GARP , P30279 , P07332 , P04626 , P01127 , and Q05066 . The highest frequencies of losses were detected in p44S10 , O15164 , P06858 , Q13126 , P35226 , P11161 , and Q13163 . Genomic alterations in TGFbeta2 , cellular retinoid-binding protein 1 gene ( P09455 ) , P42336 , P28222 , P01112 , P21860 , and O14965 differed significantly between primary OSCC and their metastatic counterparts . Genomic alterations in Q05513 , P00519 , and P08620 were significantly different in patients who died compared with those who survived . Immunohistochemistry confirmed high P42336 immunoreactivity in primary and metastatic OSCC . Higher P08620 immunoreactivity in primary OSCC is associated with a worse prognosis . Loss of P09455 immunoreactivity is evident in primary and metastatic OSCC . Our study suggests that precise genomic profiling can be useful in determining gene number changes in OSCC . As our understanding of these changes grow , this profiling may become a practical tool for clinical evaluation . Two-dimensional liquid crystalline growth within a phase-field-crystal model . By using a two-dimensional phase-field-crystal ( P27918 ) model , the liquid crystalline growth of the plastic triangular phase is simulated with emphasis on crystal shape and topological defect formation . The equilibrium shape of a plastic triangular crystal ( PTC ) grown from an isotropic phase is compared with that grown from a columnar or smectic-A ( Q13216 ) phase . While the shape of a PTC nucleus in the isotropic phase is almost identical to that of the classical P27918 model , the shape of a PTC nucleus in Q13216 is affected by the orientation of stripes in the Q13216 phase , and irregular hexagonal , elliptical , octagonal , and rectangular shapes are obtained . Concerning the dynamics of the growth process , we analyze the topological structure of the nematic order , which starts from nucleation of +1/2 and -1/2 disclination pairs at the PTC growth front and evolves into hexagonal cells consisting of +1 vortices surrounded by six satellite -1/2 disclinations . It is found that the orientational and the positional order do not evolve simultaneously ; the orientational order evolves behind the positional order , leading to a large transition zone , which can span over several lattice spacings .

MH_train_93

MH_train_93

MH_train_93

interacts_with DB00973?

multiple_choice

Pathophysiological factors affecting CAR gene expression . The body defends itself against potentially harmful compounds , such as drugs and toxic endogenous compounds and their metabolites , by inducing the expression of enzymes and transporters involved in their metabolism and elimination . The orphan nuclear receptor CAR ( Q14994 controls phase I ( CYP2B , CYP2C , CYP3A ) , phase II ( P22309 ) , and transporter ( Q9Y6L6 , Q92887 ) genes involved in drug metabolism and bilirubin clearance . Q14994 ( CAR ) is activated by xenobiotics , such as phenobarbital , but also by toxic endogenous compounds such as bilirubin metabolite(s) . To better understand the inter- and intravariability in drug detoxification , we studied the molecular mechanisms involved in CAR gene expression in human hepatocytes . We clearly identified CAR as a glucocorticoid receptor ( GR ) target gene , and we proposed the hypothesis of a signal transduction where the activation of GR plays a critical function in CAR-mediated cellular response . According to our model , chemicals or pathophysiological factors that affect GR function should decrease CAR function . To test this hypothesis , we recently investigated the effect of microtubule disrupting agents ( MIAs ) or proinflammatory cytokines . These compounds are well-known inhibitors of GR transactivation property . MIAs activate c-Jun N-terminal kinase ( JNK ) , which phosphorylates and inactivates GR , whereas proinflammatory cytokines , such as P05231 or IL1beta , induce AP-1 or NF-kB activation , respectively , leading to GR inhibition . As expected , we observed that these molecules inhibit both CAR gene expression and phenobarbital-mediated CYP gene expression in human hepatocytes . Nardosinone protects H9c2 cardiac cells from angiotensin II-induced hypertrophy . Pathological cardiac hypertrophy induced by angiotensin II ( AngII ) can subsequently give rise to heart failure , a leading cause of mortality . Nardosinone is a pharmacologically active compound extracted from the roots of Nardostachys chinensis , a well-known traditional Chinese medicine . In order to investigate the effects of nardosinone on AngII-induced cardiac cell hypertrophy and the related mechanisms , the myoblast cell line H9c2 , derived from embryonic rat heart , was treated with nardosinone ( 25 , 50 , 100 , and 200 μmol/L ) or AngII ( 1 μmol/L ) . Then cell surface area and mRNA expression of classical markers of hypertrophy were detected . The related protein levels in PI3K/Akt/ P42345 and MEK/ P29323 signaling pathways were examined by Western blotting . It was found that pretreatment with nardosinone could significantly inhibit the enlargement of cell surface area induced by AngII . The mRNA expression of P01160 , DB04899 and β-MHC was obviously elevated in AngII-treated H9c2 cells , which could be effectively blocked by nardosinone at the concentration of 100 μmol/L . Further study revealed that the protective effects of nardosinone might be mediated by repressing the phosphorylation of related proteins in PI3K/Akt and MEK/ P29323 signaling pathways . It was suggested that the inhibitory effect of nardosinone on Ang II-induced hypertrophy in H9c2 cells might be mediated by targeting PI3K/Akt and MEK/ P29323 signaling pathways . [ P01308 resistance and hyperlipidemia in the elderly ] . To elucidate the relationship between hyperlipidemias and insulin resistance in the elderly , we conducted three studies : 1 ) determination of the prevalence of hyperlipidemias in elderly subjects with impaired glucose tolerance or non-insulin dependent diabetes mellitus , 2 ) measurement of plasma glucose and insulin levels in patients with phyerlipidemias and atherosclerotic vascular disease , and 3 ) computation of correlation between levels of substances involved in coagulation and fibrinolysis ( F-VII , F-X , and P05121 ) and levels of triglycerides and insulin in serum in hyperlipidemic patients with atherosclerotic vascular disease . The prevalence of hypertriglyceridemia was 4 % in non-diabetic control subjects , 38 % in subjects with impaired glucose tolerance , 22 % in those with diabetes , and 29 % in those with both conditions . Levels of glucose and insulin in plasma were measured after oral intake of 75 g of glucose . The insulin response was greater in the group with hypertriglyceridemia than in the group with normal triglyceride levels , although the glucose responses did not differ between the groups . The activities and levels of F-VII , F-X , and P05121 correlated with triglycerides in serum and also with fasting insulin levels in hyperlipidemic patients with atherosclerotic vascular disease . We conclude that hypertriglyceridemia plays an important role in the development of atherothrombotic vascular disease ; it accompanies elevation of coagulation and antifibrinolytic activities in elderly patients with insulin resistance . Genetics of familial forms of thrombocytopenia . The joint application of clinical and genetic investigation to patients with inherited thrombocytopenias , as well as the availability of new methods for studying megakaryopoiesis , has greatly expanded the knowledge of these disorders in the last few years with regard to their etiology , pathogenesis and clinical aspects . In particular , new diseases have been described , as deriving from mutations in the genes P21333 , Q9H4B7 , P17301 / P05106 , Q9UPS8 , P99999 , and Q9H222 or Q9H221 . Moreover , forms previously considered separate entities were found to be different clinical aspects of a single disease . For instance , identification of P35579 as the gene whose mutations cause the May-Hegglin anomaly led to the recognition that Sebastian platelet syndrome , Epstein syndrome , and Fechtner syndrome derive from mutations of the same gene and describe overlapping disorders . Despite these advances , knowledge of hereditary thrombocytopenias is still far from satisfactory because for approximately half of the patients it is not possible to formulate a definite diagnosis in that their illnesses has not yet been described . In this review , we provide a systematic description of hereditary thrombocytopenias as we know them today , giving special attention to genetic aspects . Inhibition of phosphatidylinositol 3-kinase- and P29323 MAPK-regulated protein synthesis reveals the pro-apoptotic properties of P25942 ligation in carcinoma cells . P25942 , a member of the tumor necrosis factor receptor superfamily , is frequently expressed in carcinomas where its stimulation results in induction of apoptosis when de novo protein synthesis is inhibited . The requirement of protein synthesis inhibition for efficient killing suggests that P25942 transduces potent survival signals capable of suppressing its pro-apoptotic effects . We have found that inhibition of P25942 signaling on the phosphatidylinositol 3-kinase ( PI3K ) and P29323 MAPK but not on the p38 MAPK axis disrupts this balance and sensitizes carcinoma cells to P25942 -mediated cell death . The P25942 -mediated PI3K and P29323 activities were found to converge on the regulation of protein synthesis in carcinoma cells via a pathway involving the activation of p90 ribosomal S6 kinase ( p90Rsk ) and p70S6 kinases , upstream of the translation elongation factor eEF2 . In addition , P25942 ligation was found to mediate a PI3K- and mammalian target of rapamycin ( P42345 ) -dependent phosphorylation of Q13541 and its subsequent dissociation from the P06730 P06730 as well as an P29323 -dependent phosphorylation of P06730 , thus promoting translation initiation . Concomitantly , the antiapoptotic protein cFLIP was found to be induced in P29965 -stimulated carcinoma cells in a PI3K- , P29323 - , and mammalian target of rapamycin ( P42345 ) -dependent manner and down-regulation of cFLIPS expression sensitized to P25942 -mediated carcinoma cell death . These data underline the significance of the PI3K and P29323 pathways in controlling the balance between P25942 -mediated survival and death signals through the regulation of the protein synthesis machinery . Pharmacological agents that target this machinery or its upstream kinases could , therefore , be exploited for P25942 -based tumor therapy . [ The effect of type II diabetes and the metabolic syndrome on cardiac second window preconditioning ] . Preconditioning is the most powerful endogenous mechanism , to protect the heart against ischemic damage . Conflicting data are published whether preconditioning can be induced in case of diabetes and the metabolic syndrome , which are clinically very relevant conditions . If preconditioning could be induced consistently and chronically in this population , an important reduction of surgical morbidity and mortality could be reached . In this project we induced hypoxic preconditioning in mice and used cardiac pressure-conductance catheterisation and infarct size as outcome parameters . In the first part , we found that hypoxic preconditioning was capable to reduce infarct size with 40 % and preserve the load-independent parameters with 33 % after coronary occlusion . A DKO ( double knock-out : ob/ob ; P01130 -/- ) model for the metabolic syndrome developed a larger infarct size and had a reduced contractility . No preconditioning could be induced in this model . To detect the determing factor of the resistance to preconditioning , we used single knock-out models . A comparable preconditioning effect of wild type mice could be induced in the lipoprotein receptor deficient ( P01130 -/- ) model for dyslipidemia . The leptin deficient ( ob/ob ) model , characterized by insulin resistance and abdominal obesity had , identically to the DKO model , a larger infarct size . A second window of preconditioning could be induced , although it was less pronounced than the wild type and P01130 -/- model . P01308 resistance and abdominal obesity could be identified as the major factor in the resistance to preconditioning . [ Development of simplified and rapid detection assay for genetic polymorphisms influencing drug response and its clinical applications ] . Clinically important genetic polymorphisms influencing drug metabolism and drug response have typically been discovered on the basis of phenotypic differences among individuals from different populations . Routine genotyping before drug therapy may enable the identification of responders , nonresponders , or patients at increased risk of toxicity . Automated , high-throughput detecting methods for single-nucleotide polymorphisms ( SNPs ) are highly desirable in many clinical laboratories . The aim of this study is to develop a high-throughput genotyping method for detecting SNPs influencing drug response in the Japanese population . We have developed three real-time PCR assays for detecting SNPs in the human drug-metabolizing enzymes and drug targets . The assay for simultaneously detecting P11509 , P20813 , P11712 , P33260 , P33261 , P10635 , P05181 , P20815 , NAT2 , P51580 , Q12882 , P22309 , P05091 , P00325 , P08183 , P11597 , P12821 -1 , P07550 , P28223 , P49441 , P48061 , and mitochondrial DNA polymorphisms takes less than 1.5 h . With the clinical application of NAT2 genotyping , we found statistically significant difference between the incidence of adverse drug reactions ( ADRs ) and the NAT2 genotype . The incidence of the ADRs was significantly higher in the slow type than the in other two types , as 5 of the 6 patients were of the slowtype , and the other was the intermediatetype , while no patients of the rapidtype has developed any ADRs . DB00973 as a potential treatment for non-alcoholic fatty liver disease : is the intestine a modulator of hepatic insulin sensitivity and hepatic fat accumulation ? Non-alcoholic fatty liver disease ( NAFLD ) is the hepatic component of the metabolic syndrome and is known to be associated with marked insulin resistance and increased risk of cardiovascular disease . DB00973 , an inhibitor of intestinal cholesterol absorption , inhibits Niemann-Pick C1-like 1 ( Q9UHC9 ) . Interestingly , Q9UHC9 is abundantly expressed in human liver , as well as in the intestine . Recent reports suggest a potential benefit of ezetimibe in improving hepatic insulin sensitivity and decreasing hepatic inflammation and lipid accumulation . P01308 resistance and excess hepatic fat accumulation are regarded as key factors in the pathogenesis of NAFLD . We suggest , therefore , that urgent studies are needed to assess the potential therapeutic benefit of ezetimibe in treating NAFLD . Nuclear hormone receptors and cholesterol trafficking : the orphans find a new home . There are many homeostatic mechanisms that contribute to the regulation of cellular and serum cholesterol levels in humans . Much of our understanding of this regulation arose from studies of the cellular pathways controlling cholesterol synthesis and the uptake and degradation of low-density lipoproteins . The physiology governing cholesterol disposition in whole animals , including the molecular mechanisms responsible for dietary uptake of cholesterol and its subsequent catabolism to bile acids , are only now being uncovered . This review summarizes recent studies that have used modern genetic techniques , as well as cell biological methods , to begin to elucidate the pathways responsible for cholesterol trafficking in vivo . This work has led to the realization that networks of nuclear hormone receptors , including the LXR , Q96RI1 , Q07869 , and RXR proteins , play critical roles in lipid metabolism by virtue of their transcriptional regulation of the genes that control sterol metabolic pathways . Some of the major downstream targets of these regulatory networks involve members of the ABC transporter family , including O95477 , P45844 , Q9H222 , Q9H221 , P21439 /Mdr2 , and SPGP/ O95342 . These transporters contribute to the movement of sterols and biliary lipids across cellular membranes via mechanisms that have yet to be elucidated . The potential for manipulating these cholesterol trafficking pathways via drugs targeted to the nuclear hormone receptors has generated great interest in their biology and will undoubtedly lead to new therapeutic approaches to human disorders in the coming years . Application of HapMap data to the evaluation of 8 candidate genes for pediatric slow transit constipation . BACKGROUND : Slow transit constipation ( P52823 ) affects up to 3 % of all children and is an increasingly recognized cause of chronic constipation in children . We conducted a pilot study to investigate whether genes encoding neurotransmitters ( P20366 , Q9UHF0 , P01282 , NOS1 ) and receptors ( P25103 , P21452 , P29371 , P10721 ) could be responsible for P52823 . METHODS : One hundred seventeen tag single nucleotide polymorphisms ( SNPs ) , distributed among the candidate genes , were selected from HapMap data and genotyped using Sequenom ( San Diego , CA ) technology in 35 affected families . Evaluation of association was performed by transmission disequilibrium test and multilocus analysis . RESULTS : Five SNPs ( rs3771863 , rs4580655 , rs11722288 , rs4563545 , and rs3782221 ) in the P25103 , P29371 , P10721 , and NOS1 genes were found to be potentially associated with P52823 , although the significance of these results does not withstand correction for multiple testing . CONCLUSIONS : Our data indicate that 5 SNPs in the NOS1 , P25103 , P29371 , and P10721 genes could be involved in P52823 , especially rs3771863 in intron 1 of P25103 , which showed the highest association . Increase in proinflammatory cytokines in peripheral blood without haemostatic changes after LPS inhalation . INTRODUCTION : Bronchoalveolar fibrin deposition is a characteristic of various lung disorders including acute lung injury , acute respiratory distress syndrome and sepsis . It is secondary to the activation of coagulation and inhibition of fibrinolysis in the alveolar space , and can be stimulated by lipopolysaccharide ( LPS ) inhalation . The aim of this study was to determine the relation between compartmental stress in the lung and systemic response after LPS inhalation by measuring haemostatic parameters . PATIENTS AND METHODS : 12 healthy subjects underwent a bronchial challenge test with LPS ; sequential dosages were performed for 5 biological markers ( P05231 ( P05231 ) , C-Reactive Protein ( CRP ) , P00734 Fragments 1 and 2 ( F 1+2 ) , cortisol and P00747 Activator Inhibitor 1 ( P05121 ) before endotoxin inhalation and 2 , 4 , 6 , 8 and 24 hours afterwards . RESULTS : P05231 and CRP levels in the peripheral blood were higher after LPS inhalation ; there was no activation of coagulation and no increase in P05121 level . CONCLUSION : This study confirms that despite systemic release of proinflammatory cytokines , LPS inhalation does not induce systemic haemostatic response to LPS challenge . Apolipoprotein B100 danger-associated signal 1 ( ApoBDS-1 ) triggers platelet activation and boosts platelet-leukocyte proinflammatory responses . Low-density lipoproteins ( LDL ) , occurring in vivo in both their native and oxidative form , modulate platelet function and thereby contribute to atherothrombosis . We recently identified and demonstrated that ' ApoB100 danger-associated signal 1 ' ( ApoBDS-1 ) , a native peptide derived from P04114 ( ApoB100 ) of LDL , induces inflammatory responses in innate immune cells . Platelets are critically involved in the development as well as in the lethal consequences of atherothrombotic diseases , but whether ApoBDS-1 has also an impact on platelet function is unknown . In this study we examined the effect of ApoBDS-1 on human platelet function and platelet-leukocyte interactions in vitro . Stimulation with ApoBDS-1 induced platelet activation , degranulation , adhesion and release of proinflammatory cytokines . ApoBDS-1-stimulated platelets triggered innate immune responses by augmenting leukocyte activation , adhesion and transmigration to/through activated HUVEC monolayers , under flow conditions . These platelet-activating effects were sequence-specific , and stimulation of platelets with ApoBDS-1 activated intracellular signalling pathways , including Ca2+ , PI3K/Akt , P98160 , and p38- and P29323 -MAPK . Moreover , our data indicates that ApoBDS-1-induced platelet activation is partially dependent of positive feedback from ADP on P47900 and Q9H244 , and TxA2 . In conclusion , we demonstrate that ApoBDS-1 is an effective platelet agonist , boosting platelet-leukocyte 's proinflammatory responses , and potentially contributing to the multifaceted inflammatory-promoting effects of LDL in the pathogenesis of atherothrombosis . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . P42892 and β-arrestins exert spatiotemporal control of DB05875 -induced inflammatory signals . Although the intracellular trafficking of G protein-coupled receptors controls specific signaling events , it is unclear how the spatiotemporal control of signaling contributes to complex pathophysiological processes such as inflammation . By using bioluminescence resonance energy transfer and superresolution microscopy , we found that DB05875 ( SP ) induces the association of the neurokinin 1 receptor ( P25103 ) with two classes of proteins that regulate SP signaling from plasma and endosomal membranes : the scaffolding proteins β-arrestin ( βARRs ) 1 and 2 and the transmembrane metallopeptidases ECE-1c and ECE-1d . In HEK293 cells and non-transformed human colonocytes , we observed that G protein-coupled receptor kinase 2 and β P49407 /2 terminate plasma membrane Ca(2+) signaling and initiate receptor trafficking to endosomes that is necessary for sustained activation of ERKs in the nucleus . βARRs deliver the SP- P25103 endosomes , where P42892 associates with the complex , degrades SP , and allows the P25103 , freed from βARRs , to recycle . Thus , both P42892 and βARRs mediate the resensitization of P25103 Ca(2+) signaling at the plasma membrane . Sustained exposure of colonocytes to SP activates NF-κB and stimulates P10145 secretion . This proinflammatory signaling is unaffected by inhibition of the endosomal P29323 pathway but is suppressed by P42892 inhibition or β P32121 knockdown . Inhibition of protein phosphatase 2A , which also contributes to sustained P25103 signaling at the plasma membrane , similarly attenuates P10145 secretion . Thus , the primary function of βARRs and P42892 in SP-dependent inflammatory signaling is to promote resensitization , which allows the sustained P25103 signaling from the plasma membrane that drives inflammation . The transcriptional coactivator DRIP/mediator complex is involved in vitamin D receptor function and regulates keratinocyte proliferation and differentiation . Mediator is a multisubunit coactivator complex that facilitates transcription of nuclear receptors . We investigated the role of the mediator complex as a coactivator for vitamin D receptor ( P11473 ) in keratinocytes . Using P11473 affinity beads , the vitamin D receptor interacting protein ( DRIP ) /mediator complex was purified from primary keratinocytes , and its subunit composition was determined by mass spectrometry . The complex included core subunits , such as Q15648 /MED1 ( MED1 ) , that directly binds to P11473 . Additional subunits were identified that are components of the RNA polymerase II complex . The functions of different mediator components were investigated by silencing its subunits . The core subunit MED1 facilitates P11473 activity and regulating keratinocyte proliferation and differentiation . A newly described subunit Q13503 also has a role in promoting keratinocyte proliferation and differentiation , whereas Q9BTT4 has an inhibitory role . Blocking MED1/ Q13503 expression caused hyperproliferation of keratinocytes , accompanied by increases in mRNA expression of the cell cycle regulator cyclin D1 and/or glioma-associated oncogene homolog . Blocking MED1 or Q13503 expression also resulted in defects in calcium-induced keratinocyte differentiation , as indicated by decreased expression of differentiation markers and decreased translocation of P12830 to the membrane . These results show that keratinocytes use the transcriptional coactivator mediator to regulate P11473 functions and control keratinocyte proliferation and differentiation . Subjects with low plasma HDL cholesterol levels are characterized by an inflammatory and oxidative phenotype . BACKGROUND : Epidemiological studies have shown that low plasma levels of high-density lipoprotein ( HDL ) cholesterol are associated with increased risk of cardiovascular disease , but the mechanisms for the possible atheroprotective effects of HDL cholesterol have still not been fully clarified , in particular in relation to clinical studies . OBJECTIVE : To examine the inflammatory , anti-oxidative and metabolic phenotype of subjects with low plasma HDL cholesterol levels . METHODS AND RESULTS : Fifteen subjects with low HDL cholesterol levels ( eleven males and four females ) and 19 subjects with high HDL ( three males and 16 females ) were recruited . Low HDL cholesterol was defined as ≤10th age/sex specific percentile and high HDL-C was defined as ≥90 age/sex specific percentile . Inflammatory markers in circulation and PBMC gene expression of cholesterol efflux mediators were measured . Our main findings were : ( i ) subjects with low plasma HDL cholesterol levels were characterized by increased plasma levels of CRP , P14780 , neopterin , Q9H2A7 and P05362 as well as low plasma levels of adiponectin , suggesting an inflammatory phenotype ; ( ii ) these individuals also had reduced paraoxonase ( P27169 )1 activity in plasma and Q15165 gene expression in peripheral blood mononuclear cells ( PBMC ) accompanied by increased plasma levels of oxidized LDL suggesting decreased anti-oxidative capacity ; and ( iii ) PBMC from low HDL subjects also had decreased mRNA levels of O95477 and P45844 , suggesting impaired reverse cholesterol transport . CONCLUSION : Subjects with low plasma HDL cholesterol levels are characterized by an inflammatory and oxidative phenotype that could contribute to the increased risk of atherosclerotic disorders in these subjects with low HDL levels . DB08816 reduces neutrophil recruitment and lung damage in abdominal sepsis . Abstract Platelets play an important role in abdominal sepsis and Q9H244 receptor antagonists have been reported to exert anti-inflammatory effects . Herein , we assessed the impact of platelet inhibition with the Q9H244 receptor antagonist ticagrelor on pulmonary neutrophil recruitment and tissue damage in a model of abdominal sepsis . Wild-type C57BL/6 mice were subjected to cecal ligation and puncture ( CLP ) . Animals were treated with ticagrelor ( 100 mg/kg ) or vehicle prior to CLP induction . Edema formation and bronchoalveolar neutrophils as well as lung damage were quantified . Flow cytometry was used to determine expression of platelet-neutrophil aggregates , neutrophil activation and P29965 expression on platelets . CLP-induced pulmonary infiltration of neutrophils at 24 hours was reduced by 50 % in ticagrelor-treated animals . Moreover , ticagrelor abolished CLP-provoked lung edema and decreased lung damage score by 41 % . Notably , ticagrelor completely inhibited formation of platelet-neutrophil aggregates and markedly reduced thrombocytopenia in CLP animals . In addition , ticagrelor reduced platelet shedding of P29965 in septic mice . Our data indicate that ticagrelor can reduce CLP-induced pulmonary neutrophil recruitment and lung damage suggesting a potential role for platelet antagonists , such as ticagrelor , in the management of patients with abdominal sepsis . Targeting mitochondrial 18 kDa translocator protein ( TSPO ) regulates macrophage cholesterol efflux and lipid phenotype . The aim of the present study was to establish mitochondrial cholesterol trafficking 18 kDa translocator protein ( TSPO ) as a potential therapeutic target , capable of increasing macrophage cholesterol efflux to (apo)lipoprotein acceptors . Expression and activity of TSPO in human ( THP-1 ) macrophages were manipulated genetically and by the use of selective TSPO ligands . Cellular responses were analysed by quantitative PCR ( Q-PCR ) , immunoblotting and radiolabelling , including [3H]cholesterol efflux to (apo)lipoprotein A-I ( apoA-I ) , high-density lipoprotein ( HDL ) and human serum . Induction of macrophage cholesterol deposition by acetylated low-density lipoprotein ( AcLDL ) increased expression of TSPO mRNA and protein , reflecting findings in human carotid atherosclerosis . Transient overexpression of TSPO enhanced efflux ( E % ) of [3H]cholesterol to apoA-I , HDL and human serum compared with empty vector ( EV ) controls , whereas gene knockdown of TSPO achieved the converse . Ligation of TSPO ( using PK11195 , FGIN-1-27 and flunitrazepam ) triggered increases in [3H]cholesterol efflux , an effect that was amplified in TSPO-overexpressing macrophages . Overexpression of TSPO induced the expression of genes [ Q07869 ( peroxisome-proliferator-activated receptor α ) , Q13133 ( nuclear receptor 1H3/liver X receptor α ) , O95477 ( DB00171 -binding cassette A1 ) , Q9H172 ( DB00171 -binding cassette G4 ) and P02649 ( apolipoprotein E ) ] and proteins ( O95477 and PPARα ) involved in cholesterol efflux , reduced macrophage neutral lipid mass and lipogenesis and limited cholesterol esterification following exposure to AcLDL . Thus , targeting TSPO reduces macrophage lipid content and prevents macrophage foam cell formation , via enhanced cholesterol efflux to (apo)lipoprotein acceptors . Leptin promotes neointima formation and smooth muscle cell proliferation via NADPH oxidase activation and signalling in caveolin-rich microdomains . AIMS : P02649 ( apoE ) may act as a vasculoprotective factor by promoting plasma lipid clearance and cholesterol efflux . Moreover , apoE accumulates at sites of vascular injury and modulates the effect of growth factors on smooth muscle cells ( SMCs ) . Experimental data suggested that hypothalamic apoE expression is reduced in obesity and associated with leptin resistance . In this study , we examined the role of apoE in mediating the effects of leptin on vascular lesion formation . METHODS AND RESULTS : Leptin was administered to apoE knockout ( apoE-/- ) mice via osmotic pumps to increase its circulating levels . Morphometric analysis revealed that leptin did not alter neointima formation and failed to increase α-actin- or P12004 -immunopositive SMCs after vascular injury . Similar findings were obtained after analysis of atherosclerotic lesions . Comparison of apoE-/- , wild-type , or P01130 -/- mice and functional analyses in aortic SMCs from WT or apoE-/- mice or human arterial SMCs after treatment with small interfering (si)RNA or heparinase revealed that leptin requires the presence of apoE , expressed , secreted and bound to the cell surface , to fully activate leptin receptor signalling and to promote SMC proliferation and neointima formation . Mechanistically , leptin induced the phosphorylation and membrane translocation of caveolin (cav)-1 , and apoE down-regulation or caveolae disruption inhibited the leptin-induced p47phox activation , ROS formation and SMC proliferation . Finally , leptin failed to increase neointima formation in mice lacking cav-1 . CONCLUSION : Our findings suggest that apoE mediates the effects of leptin on vascular lesion formation by stabilizing cav-1-enriched cell membrane microdomains in SMCs , thus allowing NADPH oxidase assembly and ROS-mediated mitogenic signalling . Effect of canagliflozin on renal threshold for glucose , glycemia , and body weight in normal and diabetic animal models . BACKGROUND : DB08907 is a sodium glucose co-transporter ( SGLT ) 2 inhibitor in clinical development for the treatment of type 2 diabetes mellitus ( T2DM ) . METHODS : (14)C-alpha-methylglucoside uptake in Chinese hamster ovary-K cells expressing human , rat , or mouse SGLT2 or P13866 ; (3)H-2-deoxy-d-glucose uptake in Q9BTT4 myoblasts ; and 2-electrode voltage clamp recording of oocytes expressing human SGLT3 were analyzed . Graded glucose infusions were performed to determine rate of urinary glucose excretion ( UGE ) at different blood glucose ( BG ) concentrations and the renal threshold for glucose excretion ( RT(G) ) in vehicle or canagliflozin-treated Zucker diabetic fatty ( ZDF ) rats . This study aimed to characterize the pharmacodynamic effects of canagliflozin in vitro and in preclinical models of T2DM and obesity . RESULTS : Treatment with canagliflozin 1 mg/kg lowered RT(G) from 415±12 mg/dl to 94±10 mg/dl in ZDF rats while maintaining a threshold relationship between BG and UGE with virtually no UGE observed when BG was below RT(G) . DB08907 dose-dependently decreased BG concentrations in db/db mice treated acutely . In ZDF rats treated for 4 weeks , canagliflozin decreased glycated hemoglobin ( HbA1c ) and improved measures of insulin secretion . In obese animal models , canagliflozin increased UGE and decreased BG , body weight gain , epididymal fat , liver weight , and the respiratory exchange ratio . CONCLUSIONS : DB08907 lowered RT(G) and increased UGE , improved glycemic control and beta-cell function in rodent models of T2DM , and reduced body weight gain in rodent models of obesity . Oxysterol binding protein induces upregulation of SREBP-1c and enhances hepatic lipogenesis . BACKGROUND : Oxysterol binding protein ( P22059 ) has previously been implicated as a sterol sensor that regulates sphingomyelin synthesis and the activity of extracellular signal-regulated kinases ( P29323 ) . METHODS AND RESULTS : We determined the effects of adenovirus-mediated hepatic overexpression of P22059 and its homologues ORP1L and Q9H4L5 on mouse serum lipids . Whereas ORP1L and Q9H4L5 had no effect on serum lipids , P22059 induced a marked increase of VLDL triglycerides ( TG ) . Also , the liver tissue TG were elevated in the AdOSBP-injected mice , and their TG secretion rate was increased by 70 % . The messenger RNAs for enzymes of fatty acid synthesis and their transcriptional regulator , SREBP-1c , as well as the Insig-1 mRNA , were upregulated two-fold in the P22059 -expressing livers . No change occurred in the messages of liver X receptor target genes O95477 , Q9H222 , and P22680 , and the Insig-2a mRNA was reduced . The phosphorylation of P29323 was decreased in AdOSBP-infected liver and cultured hepatocytes . Importantly , silencing of P22059 in hepatocytes suppressed the induction of P36956 -c by insulin and resulted in a reduction of TG synthesis . CONCLUSION : Our results demonstrate that P22059 regulates hepatic TG metabolism and suggest the involvement of P22059 in the insulin signaling pathways that control hepatic lipogenesis . Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in Q9BTT4 rat myotubes : role of mangiferin . BACKGROUND AND AIMS : To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal alpha-glycosidases but also by enhancing glucose transport in muscle and adipose cells . METHODS : S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the amount and translocation of glucose transporters were assayed . A fractionation of the extract was carried out to identify the active compounds . Furthermore , we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake . RESULTS : S. oblonga extract increased 2-deoxy-D-glucose uptake by 50 % in Q9BTT4 -myotubes and 3T3-adipocytes . In Q9BTT4 -myotubes , the extract increased up to a 100 % the P14672 content , activating P14672 promoter transcription and its translocation to the plasma membrane . Mangiferin was identified as the bioactive compound . Furthermore , mangiferin effects were concomitant with the phosphorylation of DB00131 -activated protein kinase without the activation of P31749 /Akt . The effect of mangiferin on 2-deoxy-D-glucose uptake was blocked by GW9662 , an irreversible P37231 antagonist . CONCLUSIONS : S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing P14672 expression and translocation in muscle cells . These effects are probably mediated through two independent pathways that are related to DB00131 -activated protein kinase and P37231 . Agonists and antagonists for P2 receptors . Recent work has identified nucleotide agonists selective for P47900 , P41231 and Q15077 receptors and nucleotide antagonists selective for P47900 , Q9H244 and P51575 receptors . Selective non-nucleotide antagonists have been reported for P47900 , P41231 , Q15077 , Q9H244 , Q9BPV8 , P2X(2/3)/ P56373 and Q99572 receptors . For example , the dinucleotide P01308 37217 ( Up4dC ) potently activates the P41231 receptor , and the non-nucleotide antagonist A-317491 is selective for P2X(2/3)/ P56373 receptors . Nucleotide analogues in which the ribose moiety is substituted by a variety of novel ring systems , including conformationally locked moieties , have been synthesized as ligands for P2Y receptors . The focus on conformational factors of the ribose-like moiety allows the inclusion of general modifications that lead to enhanced potency and selectivity . At P47900 ,2,4,11 receptors , there is a preference for the North conformation as indicated with ( N ) -methanocarba analogues . The P47900 antagonist MRS2500 inhibited ADP-induced human platelet aggregation with an IC50 of 0.95 nM . MRS2365 , an ( N ) -methanocarba analogue of 2-MeSADP , displayed potency ( EC50 ) of 0.4nM at the P47900 receptor , with > 10000-fold selectivity in comparison to Q9H244 and Q9BPV8 receptors . At Q15077 receptors there is a dramatic preference for the South conformation . Three-dimensional structures of P2Y receptors have been deduced from structure activity relationships ( SAR ) , mutagenesis and modelling studies . Detailed three-dimensional structures of P2X receptors have not yet been proposed . Effects of peroxisome proliferator-activated receptor ligands , bezafibrate and fenofibrate , on adiponectin level . OBJECTIVE : Q15848 is adipose-specific secretory protein and acts as anti-diabetic and anti-atherosclerotic molecule . We previously found peroxisome proliferators response element in adiponectin promoter region , suggesting that peroxisome proliferator-activated receptor ( Q07869 ) ligands elevate adiponectin . Fibrates are known to be PPARalpha ligands and were shown to reduce risks of diabetes and cardiovascular disease . Effect of fibrates on adiponectin has not been clarified , whereas thiazolidinediones enhance adiponectin . Thus , we explored the possibility and mechanism that fibrates enhance adiponectin in humans , mice , and cells . METHODS AND RESULTS : Significant increase of serum adiponectin was observed in bezafibrate-treated subjects compared with placebo group in patients enrolled in The DB01393 Infarction Prevention study . Higher baseline adiponectin levels were strongly associated with reduced risk of new diabetes . Fibrates , bezafibrate and fenofibrate , significantly elevated adiponectin levels in wild-type mice and 3T3- Q9NUQ9 adipocytes . Such an effect was not observed in PPARalpha-deficient mice and adipocytes . Fibrates activated adiponectin promoter but failed to enhance its activity when the point mutation occurred in peroxisome proliferators response element site and the endogenous PPARalpha was knocked down by PPARalpha-RNAi . CONCLUSIONS : Our results suggest that fibrates enhance adiponectin partly through adipose PPARalpha and measurement of adiponectin might be a useful tool for searching subjects at high risk for diabetes . Platelet hyperreactivity explains the bleeding abnormality and macrothrombocytopenia in a murine model of sitosterolemia . Sitosterolemia is a rare , autosomal recessive disease caused by mutations in the adenosine triphosphate-binding cassette transporter genes Q9H222 or Q9H221 that result in accumulation of xenosterols in the body . Clinical manifestations include tendon xanthomas , premature coronary artery disease , hemolytic anemia , macrothrombocytopenia , and bleeding . Although the effect of sterol accumulation on the predisposition for atherosclerosis is evident , how xenosterol accumulation leads to defects in platelet physiology is unknown . Sitosterolemia induced in Abcg5- and Abcg8-deficient mice fed a high plant sterol diet resulted in accumulation of free sterols in platelet plasma membranes , leading to hyperactivatable platelets characterized by constitutive binding of fibrinogen to its αIIbβ3 integrin receptor , internalization of the αIIbβ3 complex , generation of platelet-derived microparticles , and changes in the quantity and subcellular localization of filamin . The latter was associated with macrothrombocytopenia , shedding of GPIbα , impaired platelet adhesion to P04275 , and inability to form stable thrombi . Plasma levels of soluble GPIbα were strongly correlated with plasma sitosterol levels in samples from human sitosterolemic patients , implicating a similar mechanism of sterol-induced platelet passivation in the human disease . Intercalation of plant sterols into the plasma membrane therefore results in dysregulation of multiple platelet activation pathways , leading to macrothrombocytopenia and bleeding . P32189 deficiency alters expression of genes involved in lipid metabolism , carbohydrate metabolism , and insulin signaling . P32189 ( GK ) is at the interface of fat and carbohydrate metabolism and has been implicated in insulin resistance and type 2 diabetes mellitus . To define GK 's role in insulin resistance , we examined gene expression in brown adipose tissue in a glycerol kinase knockout ( KO ) mouse model using microarray analysis . Global gene expression profiles of KO mice were distinct from wild type with 668 differentially expressed genes . These include genes involved in lipid metabolism , carbohydrate metabolism , insulin signaling , and insulin resistance . Real-time polymerase chain reaction analysis confirmed the differential expression of selected genes involved in lipid and carbohydrate metabolism . PathwayAssist analysis confirmed direct and indirect connections between glycerol kinase and genes in lipid metabolism , carbohydrate metabolism , insulin signaling , and insulin resistance . Network component analysis ( P40199 ) showed that the transcription factors ( TFs ) P37231 , P36956 , Q12772 , P40763 , P42229 , SP1 , CEBPalpha , CREB , GR and Q07869 have altered activity in the KO mice . P40199 also revealed the individual contribution of these TFs on the expression of genes altered in the microarray data . This study elucidates the complex network of glycerol kinase and further confirms a possible role for glycerol kinase deficiency , a simple Mendelian disorder , in insulin resistance , and type 2 diabetes mellitus , a common complex genetic disorder . DB04540 -lowering effect of ezetimibe in uridine diphosphate glucuronosyltransferase 1A-deficient ( Gunn ) rats . DB00973 ( EZE ) selectively blocks intestinal cholesterol absorption by interacting with Niemann-Pick C1 Like 1 ( Q9UHC9 ) . After administration , EZE is extensively metabolized in liver and intestine to its phenolic glucuronide form ( EZE-G ) by uridine diphosphate glucuronosyltransferases ( UGTs ) , among which P22309 and 1A3 exhibit highest activity . EZE-G is excreted into bile and undergoes extensive enterohepatic recirculation . Considering the pharmacokinetic properties of EZE and an in vitro binding study showing the high affinity binding of EZE-G to Q9UHC9 , glucuronidation by UGTs has been believed to be essential for the pharmacological efficacy of EZE . To study the role of glucuronidation by UGTs for the cholesterol-lowering effect of EZE , in vitro and in vivo studies were performed using Gunn rats , which hereditarily lack the expression of P22309 enzymes . The biliary excreted amount of EZE-G was reduced by 73 % up to 3 h after administration of EZE ( 0.3 mg/kg ) in Gunn rats , which is consistent with the reduction of in vitro EZE glucuronidation activity found in liver and intestinal microsome from Gunn rats . These results indicate that the formation of EZE-G in Gunn rats is much lower than that in Wistar rats . However , in vivo study showed that 0.3 mg/kg EZE , which is the clinically relevant dose , reduced cholesterol absorption in both Wistar and Gunn rats to nearly the same degree and the dose dependence was not significantly different between Wistar and Gunn rats at the range 0.001 approximately 0.3 mg/kg . These results indicate that a deficiency of P22309 activity does not necessarily alter the cholesterol-lowering effect of EZE in rats at therapeutic doses . The Q9H222 Q9H221 sterol transporter and phytosterols : implications for cardiometabolic disease . PURPOSE OF REVIEW : This review summarizes recent developments in the activity , regulation , and physiology of the Q9H222 Q9H221 ( G5G8 ) transporter and the use of its xenobiotic substrates , phytosterols , as cholesterol lowering agents in the treatment of cardiovascular disease . Recent progress has significant implications for the role of G5G8 and its substrates in complications associated with features of the metabolic syndrome . RECENT FINDINGS : Recent reports expand the clinical presentation of sitosterolemia to include platelet and adrenal dysfunction . The G5G8 sterol transporter is critical to hepatobiliary excretion of cholesterol under nonpathological conditions and has been linked to the cholesterol gallstone susceptibility . Finally , the cardiovascular benefits of cholesterol lowering through the use of phytosterol supplements were offset by vascular dysfunction , suggesting that alternative strategies to reduced cholesterol absorption offer greater benefit . SUMMARY : P01308 resistance elevates G5G8 and increases susceptibility to cholesterol gallstones . However , this transporter is critical for the exclusion of phytosterols from the absorptive pathways in the intestine . Challenging the limits of this protective mechanism through phytosterol supplementation diminishes the cardioprotective benefits of cholesterol lowering in mouse models of cardiovascular disease . [ Role of neurokinin-1 receptor in lung injury in rats with acute necrotizing pancreatitis ] . OBJECTIVE : To investigate the expression of neurokinin-1 receptor ( P25103 ) in the lung tissue , and the relationship between expression of P25103 and lung injury in rats with acute necrotizing pancreatitis ( P01160 ) . METHODS : One hundred and twenty adult Sprague-Dawley rats were randomly divided into P01160 and control groups . Animals in group P01160 were induced by the retrograde intraductal infusion of 5 % sodium taurocholate ( 0.1 ml/kg ) , and animals in normal control group received laparotomy only . The accumulation of polymorphonuclear leukocytes in lung tissues was measured with myeloperoxidase ( P05164 ) assay . Lung endothelial barrier destruction was measured by lung capillary permeability ( LCP ) . Reverse transcription polymerase chain reaction ( RT-PCR ) was used to determine the mRNA expression of P25103 , western blot analysis was used to determine P25103 protein expression levels , and immunohistochemistry was used to localize expression site of P25103 . RESULTS : P25103 mRNA level was enhanced in the lung of P01160 compared with normal control group . Western blot analysis showed overexpression of P25103 protein level exited in P01160 group . Statistical analysis revealed correlation between P25103 mRNA and P05164 ( r=0.83 , P < 0.01 ) and LCP ( r=0.79 , P < 0.01 ) respectively . With immunohistochemistry staining , moderate to strong P25103 immunoreactivity was localized to alveolar membrane , I epithelium , II epithelium and polymorphonuclear leukocytes in the lung of P01160 . CONCLUSION : In P01160 , overexpression of P25103 contributes to disturbance of neuropeptides loop , resulting in aggregation of neutrophilic granulocyte and promoting deterioration of lung injury . HIV protease inhibitors promote atherosclerotic lesion formation independent of dyslipidemia by increasing P16671 -dependent cholesteryl ester accumulation in macrophages . Protease inhibitors decrease the viral load in HIV patients , however the patients develop hypertriglyceridemia , hypercholesterolemia , and atherosclerosis . It has been assumed that protease inhibitor-dependent increases in atherosclerosis are secondary to the dyslipidemia . Incubation of THP-1 cells or human PBMCs with protease inhibitors caused upregulation of P16671 and the accumulation of cholesteryl esters . The use of P16671 -blocking antibodies , a P16671 morpholino , and monocytes isolated from P16671 null mice demonstrated that protease inhibitor-induced increases in cholesteryl esters were dependent on P16671 upregulation . These data led to the hypothesis that protease inhibitors induce foam cell formation and consequently atherosclerosis by upregulating P16671 and cholesteryl ester accumulation independent of dyslipidemia . Studies with P01130 null mice demonstrated that low doses of protease inhibitors induce an increase in the level of P16671 and cholesteryl ester in peritoneal macrophages and the development of atherosclerosis without altering plasma lipids . Furthermore , the lack of P16671 protected the animals from protease inhibitor-induced atherosclerosis . Finally , ritonavir increased P37231 and P16671 mRNA levels in a PKC- and P37231 -dependent manner . We conclude that protease inhibitors contribute to the formation of atherosclerosis by promoting the upregulation of P16671 and the subsequent accumulation of sterol in macrophages . Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta . BACKGROUND & AIMS : Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease ( Q9UKU7 ) . Various drugs used in the treatment of Q9UKU7 , such as glucocorticoids , DB00244 , and sulfasalazine , interfere with NF-kappaB/Rel signaling . The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation . METHODS : The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift , transfection , and immune complex kinase assays . The direct effect of sulfasalazine on O15111 ( IKK ) activity was investigated using purified recombinant O15111 and -beta proteins . RESULTS : NF-kappaB/Rel activity induced by tumor necrosis factor alpha , 12-O-tetradecanoylphorbol-13-acetate , or overexpression of NF-kappaB-inducing kinase , O15111 , O14920 , or constitutively active O15111 and O14920 mutants was inhibited dose dependently by sulfasalazine . Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells , as well as the catalytic activity of purified O15111 and O14920 in vitro . In contrast , the moieties of sulfasalazine , DB00244 , and sulfapyridine or DB00233 had no effect . Activation of extracellular signal-related kinase ( P29323 ) 1 and 2 , c-Jun-N-terminal kinase ( JNK ) 1 , and p38 was unaffected by sulfasalazine . The decrease in substrate phosphorylation by O15111 and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate . CONCLUSIONS : These data identify sulfasalazine as a direct inhibitor of O15111 and -beta by antagonizing adenosine triphosphate binding . The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine . Current therapy for patients with sitosterolemia -- effect of ezetimibe on plant sterol metabolism . Sitosterolemia is a rare , autosomal recessive inherited sterol storage disease associated with high tissue and serum plant sterol concentrations , caused by mutations in the adenosine triphosphate-bind-ing cassette ( DB01048 ) transporter Q9H222 or Q9H221 genes . Markedly increased serum concentration of plant sterols. such as sitosterol and campesterol , cause premature atherosclerosis and massive xanthomas . Hitherto known treatments for sitosterolemia , including a low-sterol diet , bile-salt binding resins , ileal bypass surgery and low density lipoprotein ( LDL ) apheresis have not yielded sufficient reduction of serum plant sterol levels and many patients show a sustained elevation of plant sterol levels , subsequently developing premature atherosclerotic cardiovascular diseases . DB00973 , an inhibitor of intestinal cholesterol absorption through its binding to Niemann-Pick C1-like 1 ( Q9UHC9 ) , has been widely used for decreasing serum LDL-cholesterol levels in patients with hypercholesterolemia . DB00973 also reduces the gastrointestinal absorption of plant sterols , thereby also lowering the serum concentrations of plant sterols . This pharmacological property of ezetimibe shows its potential as a novel effective therapy for sitosterolemia . In the current review , we discuss the current therapy for patients with sitosterolemia and present two Japanese adolescent patients with this disease , one of whom underwent percutaneous coronary intervention for accelerated coronary atherosclerosis . DB00973 administration in addition to conventional drug therapy successfully reduced serum sitosterol levels by 51.3 % and 48.9 % , respectively , in the two patients , demonstrating ezetimibe as a novel and potent treatment agent for sitosterolemia that could work additively with conventional drug therapy . LXR driven induction of HDL-cholesterol is independent of intestinal cholesterol absorption and O95477 protein expression . We investigated whether : ( 1 ) liver X receptor ( LXR ) -driven induction of high-density lipoprotein cholesterol ( HDL-C ) and other LXR-mediated effects on cholesterol metabolism depend on intestinal cholesterol absorption ; and ( 2 ) combined treatment with the LXR agonist GW3965 and the cholesterol absorption inhibitor ezetimibe results in synergistic effects on cholesterol metabolism that could be beneficial for treatment of atherosclerosis . Mice were fed 0.2 % cholesterol and treated with GW3965+ezetimibe , GW3965 or ezetimibe . GW3965+ezetimibe treatment elevated serum HDL-C and Apolipoprotein ( Apo ) AI , effectively reduced the intestinal cholesterol absorption and increased the excretion of faecal neutral sterols . No changes in intestinal DB00171 -binding cassette ( DB01048 ) A1 or Q9H222 protein expression were observed , despite increased mRNA expression , while hepatic O95477 was slightly reduced . The combined treatment caused a pronounced down-regulation of intestinal Niemann-Pick C1-like 1 ( Q9UHC9 ) and reduced hepatic and intestinal cholesterol levels . GW3965 did not affect the intestinal cholesterol absorption , but increased serum HDL-C and ApoAI levels . GW3965 also increased Apoa1 mRNA levels in primary mouse hepatocytes and HEPA1-6 cells . DB00973 reduced the intestinal cholesterol absorption , O95477 and Q9H222 , but did not affect the serum HDL-C or ApoAI levels . Thus , the LXR-driven induction of HDL-C and ApoAI was independent of the intestinal cholesterol absorption and increased expression of intestinal or hepatic O95477 was not required . Inhibited influx of cholesterol via Q9UHC9 and/or low levels of intracellular cholesterol prevented post-transcriptional expression of intestinal O95477 and Q9H222 , despite increased mRNA levels . Combined LXR activation and blocked intestinal cholesterol absorption induced effective faecal elimination of cholesterol . P35568 regulation in health and disease . The global incidence of diabetes is increasing at epidemic rates . Estimates suggest there are currently 150 million people with diabetes and this number is expected to double in the next 20 years . Type 2 diabetes accounts for 95 % of all cases and is characterized in part by impaired sensitivity to insulin or ' insulin resistance ' . Defects in the insulin signalling pathways underpin this resistance . In the current article we discuss the regulation of P01308 Receptor Substrate-1 ( P35568 ) , a protein that plays a pivotal role in insulin signalling and whose function is impaired in subjects with insulin resistance . Coordination of P35568 function is multi-faceted , involving phosphorylation of P35568 at multiple serine/threonine residues . This controls many aspects of P35568 , including its interaction with the insulin receptor and subsequent tyrosine phosphorylation , as well as its subcellular distribution and targeting for degradation by the proteasome . Such tight control ensures appropriate transduction and attenuation of the insulin signal , thereby regulating insulin action in healthy individuals . Emerging evidence indicates that ' diabetogenic factors ' associated with insulin resistance , such as TNFalpha and elevated circulating fatty acids , impact on insulin signalling at the level of P35568 serine/threonine phosphorylation . The expression and/or activity of several kinases , such as O15111 beta ( IKKbeta ) and salt-induced kinase 2 ( Q9H0K1 ) , and the phosphorylation of P35568 at key sites , such as Ser307 and Ser789 , are increased in states of insulin resistance . Identifying the pathways by which such factors activate these and other kinases , and defining the precise roles of specific serine/ threonine phosphorylation events in P35568 regulation , represent important goals which may eventually provide a rationale for therapeutic intervention . Q07869 and P37231 activators induce cholesterol removal from human macrophage foam cells through stimulation of the O95477 pathway . Peroxisome proliferator-activated receptors ( PPARs ) are nuclear receptors that regulate lipid and glucose metabolism and cellular differentiation . Q07869 and P37231 are both expressed in human macrophages where they exert anti-inflammatory effects . The activation of Q07869 may promote foam-cell formation by inducing expression of the macrophage scavenger receptor P16671 . This prompted us to investigate the influence of different Q07869 -activators on cholesterol metabolism and foam-cell formation of human primary and THP-1 macrophages . Here we show that Q07869 and P37231 activators do not influence acetylated low density lipoprotein-induced foam-cell formation of human macrophages . In contrast , Q07869 and P37231 activators induce the expression of the gene encoding O95477 , a transporter that controls apoAI-mediated cholesterol efflux from macrophages . These effects are likely due to enhanced expression of liver-x-receptor alpha , an oxysterol-activated nuclear receptor which induces O95477 -promoter transcription . Moreover , Q07869 and P37231 activators increase apoAI-induced cholesterol efflux from normal macrophages . In contrast , Q07869 or P37231 activation does not influence cholesterol efflux from macrophages isolated from patients with Tangier disease , which is due to a genetic defect in O95477 . Here we identify a regulatory role for Q07869 and P37231 in the first steps of the reverse-cholesterol-transport pathway through the activation of O95477 -mediated cholesterol efflux in human macrophages . Efficacy and safety of repeated dosing of netupitant , a neurokinin-1 receptor antagonist , in treating overactive bladder . AIM : NK-1 receptors in sensory nerves , the spinal cord and bladder smooth muscle participate in complex sensory mechanisms that regulate bladder activity . This study was designed to assess the efficacy and safety of a new P25103 antagonist , netupitant , in patients with OAB . METHODS : This was a phase II , multicenter , double-blind study in which adults with OAB symptoms > 6 months were randomized to receive 1 of 3 doses of netupitant ( 50 , 100 , 200 mg ) or placebo once daily for 8 weeks . The primary efficacy endpoint was percentage change from baseline in average number of daily micturitions at week 8 . Urinary incontinence , urge urinary incontinence ( UUI ) , and urgency episodes were also assessed . RESULTS : The primary efficacy endpoint was similar in the treatment groups ( -13.85 for placebo to -16.17 in the netupitant 200 mg group ) with no statistically significant differences between netupitant and placebo . The same was true for most secondary endpoints although a significant difference for improvement in UUI episodes and a trend for the greatest decrease in urgency episodes were seen in the netupitant 100 mg group . DB09048 was well tolerated with most treatment emergent adverse events ( AEs ) being mild . While the overall incidence of AEs increased with netupitant dose , there was no evidence for this dose dependency based on relationship to treatment , intensity , or time to onset . CONCLUSIONS : The study failed to demonstrate superiority of netupitant versus placebo in decreasing OAB symptoms , despite a trend favoring netupitant 100 mg . There were no safety concerns with daily administration of netupitant over 8 weeks . The molecular mechanisms underlying the reduction of LDL apoB-100 by ezetimibe plus simvastatin . The combination of ezetimibe , an inhibitor of Niemann-Pick C1-like 1 protein ( Q9UHC9 ) , and an P04035 inhibitor decreases cholesterol absorption and synthesis . In clinical trials , ezetimibe plus simvastatin produces greater LDL-cholesterol reductions than does monotherapy . The molecular mechanism for this enhanced efficacy has not been defined . P04114 ( apoB-100 ) kinetics were determined in miniature pigs treated with ezetimibe ( 0.1 mg/kg/day ) , ezetimibe plus simvastatin ( 10 mg/kg/day ) , or placebo ( n = 7/group ) . DB00973 decreased cholesterol absorption ( -79 % ) and plasma phytosterols ( -91 % ) , which were not affected further by simvastatin . DB00973 increased plasma lathosterol ( +65 % ) , which was prevented by addition of simvastatin . The combination decreased total cholesterol ( -35 % ) and LDL-cholesterol ( -47 % ) . VLDL apoB pool size decreased 26 % , due to a 35 % decrease in VLDL apoB production . LDL apoB pool size decreased 34 % due to an 81 % increase in the fractional catabolic rate , both of which were significantly greater than monotherapy . Combination treatment decreased hepatic microsomal cholesterol ( -29 % ) and cholesteryl ester ( -65 % ) and increased P01130 ( P01130 ) expression by 240 % . The combination increased Q9UHC9 expression in liver and intestine , consistent with increased Q12772 expression . DB00973 plus simvastatin decreases VLDL and LDL apoB-100 concentrations through reduced VLDL production and upregulation of P01130 -mediated LDL clearance . Inhibition of Niemann-Pick-type C1-like1 by ezetimibe activates autophagy in human hepatocytes and reduces mutant α1-antitrypsin Z deposition . Autophagy can degrade aggregate-prone proteins , but excessive autophagy can have adverse effects . It would be beneficial if autophagy could be enhanced in a cell type-specific manner , but this has been difficult because the basic mechanism of autophagy is common . In the present study we found that inhibition of Niemann-Pick-type C1-like 1 ( Q9UHC9 ) by ezetimibe activates autophagy only in hepatocytes and small intestinal epithelia , but not in other cells . DB00973 induced accumulation of free cholesterol in the late endosome/lysosome and increased partitioning of a Ragulator component , Q6IAA8 , in rafts . The latter change led to down-regulation of mammalian target of rapamycin ( P42345 )C1 activity by decreasing P42345 recruitment to the late endosome/lysosome and activated autophagy . A primary effect of ezetimibe was found to be a decrease of free cholesterol in the plasma membrane , because all the results caused by ezetimibe were suppressed by supplementation of cholesterol as a methyl-β-cyclodextrin complex . By enhancing autophagy in human primary hepatocytes with ezetimibe , insoluble mutant α1-antitrypsin Z was reduced significantly . CONCLUSION : Inhibition of Q9UHC9 by ezetimibe activates autophagy in human hepatocytes by modulating cholesterol homeostasis . DB00973 may be used to ameliorate liver degeneration in α1-antitrypsin deficiency . Genetic demonstration of intestinal Q9UHC9 as a major determinant of hepatic cholesterol and blood atherogenic lipoprotein levels . OBJECTIVE : The correlation between intestinal cholesterol absorption values and plasma low-density lipoprotein-cholesterol ( LDL-C ) levels remains controversial . Niemann-Pick-C1-Like 1 ( Q9UHC9 ) is essential for intestinal cholesterol absorption , and is the target of ezetimibe , a cholesterol absorption inhibitor . However , studies with Q9UHC9 knockout mice or ezetimibe can not definitively clarify this correlation because Q9UHC9 expression is not restricted to intestine in humans and mice . In this study we sought to genetically address this issue . METHODS AND RESULTS : We developed a mouse model that lacks endogenous ( Q9UHC9 ) and P01130 ( P01130 ) ( DKO ) , but transgenically expresses human Q9UHC9 in gastrointestinal tract only ( DKO/ Q9NUQ9 (IntOnly) mice ) . Our novel model eliminated potential effects of non-intestinal Q9UHC9 on cholesterol homeostasis . We found that human Q9UHC9 was localized at the intestinal brush border membrane of DKO/ Q9NUQ9 (IntOnly) mice . DB04540 feeding induced formation of Q9UHC9 -positive vesicles beneath this membrane in an ezetimibe-sensitive manner . Compared to DKO mice , DKO/ Q9NUQ9 (IntOnly) mice showed significant increases in cholesterol absorption and blood/hepatic/biliary cholesterol . Increased blood cholesterol was restricted to very low-density lipoprotein ( VLDL ) and LDL fractions , which was associated with increased secretion and plasma levels of apolipoproteins B100 and B48 . Additionally , DKO/ Q9NUQ9 (IntOnly) mice displayed decreased fecal cholesterol excretion and hepatic/intestinal expression of cholesterologenic genes . DB00973 treatment virtually reversed all of the transgene-related phenotypes in DKO/ Q9NUQ9 (IntOnly) mice . CONCLUSION : Our findings from DKO/ Q9NUQ9 (IntOnly) mice clearly demonstrate that Q9UHC9 -mediated cholesterol absorption is a major determinant of blood levels of apolipoprotein B-containing atherogenic lipoproteins , at least in mice . Inhibition of NF-kappaB activation in macrophages increases atherosclerosis in P01130 -deficient mice . Atherosclerosis is now generally accepted as a chronic inflammatory condition . The transcription factor NF-kappaB is a key regulator of inflammation , immune responses , cell survival , and cell proliferation . To investigate the role of NF-kappaB activation in macrophages during atherogenesis , we used P01130 -deficient mice with a macrophage-restricted deletion of O15111 2 ( O14920 ) , which is essential for NF-kappaB activation by proinflammatory signals . These mice showed increased atherosclerosis as quantified by lesion area measurements . In addition , the lesions were more advanced and showed more necrosis and increased cell number in early lesions . Southern blotting revealed that deletion of O14920 was approximately 65 % in macrophages , coinciding with a reduction of 50 % in NF-kappaB activation , as compared with controls . In both groups , the expression of differentiation markers , uptake of bacteria , and endocytosis of modified LDL was similar . Upon stimulation with LPS , production of P01375 was reduced by approximately 50 % in O14920 -deleted macrophages . Interestingly , we also found a major reduction in the anti-inflammatory cytokine P22301 . Our data show that inhibition of the NF-kappaB pathway in macrophages leads to more severe atherosclerosis in mice , possibly by affecting the pro- and anti-inflammatory balance that controls the development of atherosclerosis . DB08816 yields consistent dose-dependent inhibition of ADP-induced platelet aggregation in patients with atherosclerotic disease regardless of genotypic variations in Q9H244 , P47900 , and P05106 . The platelet P2Y(12) receptor is the target of clopidogrel therapy , which has been shown to reduce thromboembolic complications of atherosclerotic disease but has limitations in terms of variability of response and irreversibility of effect . This receptor is also a target for ticagrelor ( AZD6140 ) , the first reversibly binding oral P2Y(12) receptor antagonist that does not require metabolic activation and yields more consistent inhibition of platelet aggregation than clopidogrel therapy . Single nucleotide polymorphisms ( SNPs ) have been described in the gene for this receptor ( Q9H244 ) , some of which have been associated with variability in platelet reactivity . SNPs in P47900 and P05106 have also been reported by some groups to affect platelet reactivity to adenosine diphosphate ( ADP ) . We assessed whether SNPs in these genes influenced the pharmacodynamic response to ticagrelor in patients enrolled in both the DISPERSE study ( stable atherosclerotic disease ) and the DISPERSE2 study ( non-ST-segment elevation acute coronary syndromes ) . Platelet aggregation data ( at baseline and 4 weeks ) and DNA samples from clopidogrel-naive Caucasian patients treated with ticagrelor were available for 151 patients . Seventy four SNPs within three genes were genotyped . After adjustment for multiple comparisons , none of these SNPs were found to significantly influence inhibition of ADP-induced platelet aggregation by ticagrelor . TNFalpha induces O95477 through NF-kappaB in macrophages and in phagocytes ingesting apoptotic cells . Recent evidence suggests that tumor necrosis factor alpha ( TNFalpha ) signaling in vascular cells can have antiatherogenic consequences , but the mechanisms are poorly understood . TNFalpha is released by free cholesterol-loaded apoptotic macrophages , and the clearance of these cells by phagocytic macrophages may help to limit plaque development . Macrophage cholesterol uptake induces DB00171 -binding cassette ( DB01048 ) transporter O95477 promoting cholesterol efflux to apolipoprotein A-I and reducing atherosclerosis . We show that TNFalpha induces O95477 mRNA and protein in control and cholesterol-loaded macrophages and enhances cholesterol efflux to apolipoprotein A-I . The induction of O95477 by TNFalpha is reduced by 65 % in O15111 beta-deficient macrophages and by 30 % in p38alpha-deficient macrophages , but not in jun kinase 1 ( P45983 ) - or P45984 -deficient macrophages . To evaluate the potential pathophysiological significance of these observations , we fed TNFalpha-secreting free cholesterol-loaded apoptotic macrophages to a healthy macrophage monolayer ( phagocytes ) . O95477 mRNA and protein were markedly induced in the phagocytes , a response that was mediated both by TNFalpha signaling and by liver X receptor activation . Thus , TNFalpha signals primarily through NF-kappaB to induce O95477 expression in macrophages . In atherosclerotic plaques , this process may help phagocytic macrophages to efflux excess lipids derived from the ingestion of cholesterol-rich apoptotic corpses . 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 . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . miR-200c inhibits melanoma progression and drug resistance through down-regulation of BMI-1 . MicroRNAs ( miRNAs ) are short noncoding RNAs that play crucial roles in tumorigenesis and tumor progression . Melanoma is the most aggressive skin cancer that is resistant or rapidly develops resistance to a variety of chemotherapeutic agents . The role of miRNAs in melanoma progression and drug resistance has not been well studied . Herein , we demonstrate that miR-200c is down-regulated in melanomas ( primary and metastatic ) compared with melanocytic nevi . Overexpression of miR-200c in melanoma cells resulted in significantly decreased cell proliferation and migratory capacity as well as drug resistance . miR-200c overexpression resulted in significant down-regulation of BMI-1 , Q9UNQ0 , Q9H222 , and P08183 expression and in a concomitant increase in P12830 levels . Knockdown of BMI-1 showed similar effects as miR-200c overexpression in melanoma cells . In addition , miR-200c overexpression significantly inhibited melanoma xenograft growth and metastasis in vivo , and this correlated with diminished expression of BMI-1 and reduced levels of P12830 in these tumors . The effects of miR-200c on melanoma cell proliferation and migratory capacity and on self-renewal were rescued by overexpression of Bmi-1 , and the reversal of these phenotypes correlated with a reduction in P12830 expression and increased levels of Q9UNQ0 , Q9H222 , and P08183 . Taken together , these findings demonstrate a key role for miR-200c in melanoma progression and drug resistance . These results suggest that miR-200c may represent a critical target for increasing melanoma sensitivity to clinical therapies . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . DB00877 effects transcriptional programs in smooth muscle cells controlling proliferative and inflammatory properties . Neointima formation , the leading cause of restenosis , is caused by proliferation of coronary artery smooth muscle cells ( CASMCs ) and is associated with infiltration by monocytes . DB00877 inhibits neointima formation after stent implantation in humans . It reduces proliferation by its effects on mammalian target of rapamycin ( P42345 ) kinase . In this study , we investigated the expression of P42345 in human neointima and the effect of rapamycin on global transcriptional events controlling CASMC phenotype . In neointimal CASMCs , P42345 exhibited increased phosphorylation and was translocated to the nucleus compared with control . Comparative gene expression analysis of CASMCs treated with rapamycin ( 100 ng/ml ) revealed down-regulation of the transcription factor Q01094 , a key regulator of G(1)/S-phase entry , and of various retinoblastoma protein/ Q01094 -regulated genes . In addition , we found changes in the expression of genes associated with replication , apoptosis , and extracellular matrix formation . Furthermore , rapamycin decreased the gene expression of endothelial monocyte-activating polypeptide-II ( EMAP-II ) . This decrease of EMAP-II expression was reflected in a reduced adhesiveness of CASMCs for monocytic cells . Addition of EMAP-II counteracted the antiadhesive effect of rapamycin . Therefore , EMAP-II may comprise a mechanism of rapamycin-mediated reduction of the proinflammatory activation of CASMCs . The effects reported here of rapamycin on the down-regulation of genes involved in cell cycle progression , apoptosis , proliferation , and extracellular matrix formation in CASMCs provide an explanation of how rapamycin reduces CASMC proliferation . In addition , rapamycin may contribute to a reduction of inflammatory responses by reducing the adhesiveness of CASMC , a mechanism suggested to be mediated by the production and release of EMAP II . [ Prominent features of management strategies in acute coronary syndromes with the new oral antiplatelet agents ] . The novel oral Q9H244 inhibitors ( prasugrel and ticagrelor ) have been incorporated into the recently updated acute coronary syndrome ( ACS ) guidelines , as an adjunct antiplatelet treatment to aspirin . The studies involving the use of new oral antiplatelet agents that are more potent , predictable and faster platelet inhibitors than clopidogrel have demonstrated superiority with respect to the primary composite endpoint ( cardiovascular death , non-lethal myocardial infarction , stroke ) for both prasugrel and ticagrelor compared to clopidogrel . The subgroup analysis of the relevant studies showed that these new agents differ in their level of efficacy in different ACS patient subgroups : ( 1 ) Mortality was reduced with ticagrelor ; ( 2 ) DB08816 is especially more effective in intermediate-and high-risk non-ST elevation ACS patients in whom early invasive strategy is selected ; ( 3 ) Prasugrel should be especially preferred in patients with acute ST elevation myocardial infarction undergoing percutaneous coronary intervention ( P05154 ) after diagnostic angiography ; and ( 4 ) Prasugrel is more effective in diabetic patients . While clopidogrel is recommended for ACS patients who are followed with a non-invasive strategy or who have not undergone percutaneous revascularization , it is the last line choice or an alternative to the Q9H244 inhibitor therapy for patients undergoing invasive strategy . DB01393 restores the inhibition of DB00094 -induced follicular development and steroidogenesis by tumor necrosis factor-alpha through peroxisome proliferator-activated receptor-gamma pathway in an in vitro mouse preantral follicle culture . We recently reported that bezafibrate , a lipid-lowering drug of the fibrate class , administered in addition to clomiphene citrate ( CC ) successfully induced ovulation in CC-resistant polycystic ovary syndrome ( PCOS ) patients . We hypothesized that bezafibrate may directly affect ovarian follicle development . P01308 resistance and compensatory hyperinsulinemia are important for the pathogenesis of PCOS . In this study , we first examined the effects of tumor necrosis factor-alpha ( P01375 ) , which plays a role in insulin resistance , on follicle development by using the follicle culture system . P01375 significantly inhibited follicle-stimulating hormone ( DB00094 ) -induced follicle development , 17beta-estradiol ( E2 ) secretion , and ovulation rate in a dose-dependent manner . We then examined whether bezafibrate treatment could rescue the inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 . DB01393 treatment rescued inhibition of follicle development , secretion of E2 , and ovulation rate by P01375 . We examined the expression of peroxisome proliferator-activated receptor ( Q07869 ) subtypes in mouse preantral follicles . As the protein expression of only P37231 was observed in mouse preantral follicles , we examined whether bezafibrate could affect follicle development and steroidogenesis through P37231 pathways . Treatment with GW1929 , a selective P37231 agonist , restored inhibition of DB00094 -induced follicle development and steroidogenesis by P01375 , whereas treatment with GW9662 , a selective P37231 antagonist , canceled the restorative effects of bezafibrate . Collectively , the results in this study suggest that bezafibrate may directly exhibit a restorative effect on the inhibition of ovarian follicle development and steroidogenesis by P01375 through the P37231 pathway . Selective regulation of P22309 and SREBP-1c mRNA expression by docosahexaenoic , eicosapentaenoic , and arachidonic acids . We evaluated , in human cell line HepG2 , the action of individual dietary polyunsaturated fatty acids ( PUFAs ) on the expression of several lipid metabolism genes . The effects of docosahexaenoic acid , 22:6 , n-3 ( DB01708 ) , eicosapentaenoic acid , 20:5 , n-3 ( EPA ) , and arachidonic acid , 20:4 , n-6 ( AA ) were studied alone and with vitamin E ( Vit.E ) . DB01708 , EPA , and AA down-regulated mRNAs and encoded proteins of stearoyl- DB01992 desaturase ( O00767 ) and sterol regulatory element binding protein ( SREBP-1c ) , two major factors involved in unsaturated fatty acids synthesis . DB01708 affected SREBP-1c mRNA less markedly than EPA and AA . Vit.E did not affect these products , both when individually added or together with fatty acids . The expression of UDP-glucuronosyl transferase 1A1 ( P22309 ) mRNA , an enzyme of phase II drug metabolism with relevant actions within lipid metabolism , resulted also differentially regulated . DB01708 did not essentially reduce P22309 mRNA expression while EPA and AA produced a considerable decrease . Nevertheless , when these PUFAs were combined with Vit.E , which by itself did not produce any effect , the result was a reduction of P22309 mRNA with DB01708 , an increase reverting to basal level with EPA and no variation with AA . Observed regulations did not result to be mediated by peroxisome proliferator-activated receptor ( Q07869 ) . Our data indicate that major dietary PUFAs and Vit.E are differentially and selectively able to affect the expression of genes involved in lipid metabolism . The different actions of these slightly different molecules could be associated with their physiological role as relevant nutrient molecules . Q9Y5Q5 mutations K317E and S472G from preeclamptic patients alter zymogen activation and cell surface targeting. [ Corrected ] . Q9Y5Q5 is a membrane-bound serine protease that acts as the atrial natriuretic peptide ( P01160 ) convertase in the heart . Recent studies show that corin also activates P01160 in the pregnant uterus to promote spiral artery remodeling and prevent pregnancy-induced hypertension . Two Q9Y5Q5 gene mutations , K317E and S472G , were identified in preeclamptic patients and shown to have reduced activity in vitro . In this study , we carried out molecular modeling and biochemical experiments to understand how these mutations impair corin function . By molecular modeling , the mutation K317E was predicted to alter corin P01130 -2 module conformation . Western blot analysis of K317E mutant in HEK293 cells showed that the mutation did not block corin expression on the cell surface but inhibited corin zymogen activation . In contrast , the mutation S472G was predicted to abolish a β-sheet critical for corin frizzled-2 module structure . In Western blot analysis and flow cytometry , S472G mutant was not detected on the cell surface in transfected HEK293 cells . By immunostaining , the S472G mutant was found in the ER , indicating that the mutation S472G disrupted the β-sheet , causing corin misfolding and ER retention . Thus , these results show that mutations in the Q9Y5Q5 gene may impair corin function by entirely different mechanisms . Together , our data provide important insights into the molecular basis underlying corin mutations that may contribute to preeclampsia in patients . DB04540 and plant sterol absorption : recent insights . The recent discovery of transporters in the intestinal mucosa and the canalicular membrane has given new insights into the regulation of intestinal absorption as well as the biliary output of cholesterol and plant sterols . The 2 adenosine triphosphate ( DB00171 ) -binding cassette ( DB01048 ) half-transporters Q9H222 and Q9H221 are expressed in the mucosa cells and the canalicular membrane , and they resecrete sterols , especially absorbed plant sterols , back into the intestinal lumen and from the liver into bile . Defects of either of these cotransporters lead to the rare inherited disease of phytosterolemia , which is clinically defined by hyperabsorption and diminished biliary excretion of plant sterols . Furthermore , it has been recently demonstrated that the Niemann-Pick C1-Like 1 ( Q9UHC9 ) transporter is most likely responsible for the transport of cholesterol and plant sterols from the brush border membrane into the intestinal mucosa . DB00973 interferes with Q9UHC9 , reducing the intestinal uptake of cholesterol and plant sterols . These new findings contribute to our understanding of cholesterol and plant sterol concentrations in serum , and the effect of dietary and drug intervention to reduce serum concentrations of sterols . DB05039 inhibits tumor cell invasiveness and P14780 expression by suppressing IKK/NF-κB activation . The β2 adrenergic receptor ( P07550 ) is a G protein-coupled transmembrane receptor expressed in the human respiratory tract and widely recognized as a pharmacological target for treatments of asthma and chronic obstructive pulmonary disorder ( P48444 ) . Although a number of P07550 agonists have been developed for use in asthma therapy , indacaterol is the only ultra-long-acting inhaled β2-agonist ( LABA ) approved by the FDA for relieving the symptoms in P48444 patients . The precise molecular mechanism underlying the pharmacological effect of indacaterol , however , remains unclear . Here , we show that β-arrestin-2 mediates the internalization of P07550 following indacaterol treatment . Moreover , we demonstrate that indacaterol significantly inhibits tumor necrosis factor-α ( P01375 -α ) -induced NF-κB activity by reducing levels of both phosphorylated-IKK and -IκBα , thereby decreasing NF-κB nuclear translocation and the expression of P14780 , an NF-κB target gene . Subsequently , we show that indacaterol significantly inhibits P01375 -α/NF-κB-induced cell invasiveness and migration in a human cancer cell line . In conclusion , we propose that indacaterol may inhibit NF-κB activity in a β-arrestin2-dependent manner , preventing further lung damage and improving lung function in P48444 patients . Molecular-based choice of cancer therapy : realities and expectations . Current choice of cancer therapy is usually empirical and relies mainly on the statistical prediction of the treatment success . Molecular research provides some opportunities to personalize antitumor treatment . For example , life-threatening toxic reactions can be avoided by the identification of subjects , who carry susceptible genotypes of drug-metabolizing genes ( e.g. P51580 , P22309 , P42898 , Q12882 ) . Tumor sensitivity can be predicted by molecular portraying of targets and other molecules associated with drug response . Tailoring of antiestrogen and trastuzumab therapy based on hormone and P04626 receptor status has already become a classical example of customized medicine . Other predictive markers have been identified both for cytotoxic and for targeted therapies , and include , for example , expression of TS , TP , Q12882 , OPRT , P07992 , P16455 , P11388 , class III beta-tubulin molecules as well as genomic alterations of P00533 , P10721 , P00519 oncogenes . DB02342 causes functional repression of transforming growth factor β3 signaling by ameliorating Smad and non-Smad signaling pathways in immortalized uterine fibroid cells . OBJECTIVE : To investigate the effects and the mechanism of action of 2-methoxyestradiol ( 2ME(2) ) on transforming growth factor ( TGF ) β3-induced profibrotic response in immortalized human uterine fibroid smooth muscle ( huLM ) cells . DESIGN : Laboratory study . SETTING : University research laboratory . PATIENTS(S) : Not applicable . INTERVENTIONS(S) : Not applicable . MAIN OUTCOME MEASURE(S) : huLM cells were treated with TGF-β3 ( 5 ηg/mL ) in the presence or absence of specific P84022 inhibitor SIS3 ( 1 μmol/L ) , inhibitor of the PI3K/Akt ( LY294002 , 10 μmol/L ) , or 2ME(2) ( 0.5 μmol/L ) , and the expression of collagen ( Col ) type I(αI) , Col III(αI) , plasminogen activator inhibitor ( P05121 ) 1 , connective tissue growth factor ( P29279 ) , and α-smooth muscle actin ( α-SMA ) were determined by real-time reverse-transcription polymerase chain reaction and immunoblotting . The effect of 2ME(2) on Smad-microtubule binding was evaluated by coimmunoprecipitation . RESULT(S) : Our data revealed that TGF-β3-induced fibrogenic response in huLM is mediated by both Smad-dependent and Smad-independent PI3K/Akt/ P42345 signaling pathways . 2ME(2) abrogates TGF-β3-induced expression of Col I(αI) , Col III(αI) , P05121 , P29279 , and α-SMA . Molecularly , 2ME(2) ameliorates TGF-β3-induced Q15796 /3 phosphorylation and nuclear translocation . In addition , 2ME(2) inhibits TGF-β3-induced activation of the PI3K/Akt/ P42345 pathway . CONCLUSION(S) : TGF-β3-induced profibrotic response in fibroid cells is mediated by Smad-dependent and Smad-independent PI3K/Akt/ P42345 pathways . 2ME(2) inhibits TGF-β3 profibrotic effects in huLM cells by ameliorating both Smad-dependent and Smad-independent signaling pathways . Relative expression of cholesterol transport-related proteins and inflammation markers through the induction of 7-ketosterol-mediated stress in Caco-2 cells . Human diets contain sterol oxidation products that can induce cytotoxic effects , mainly caused by cholesterol oxides . However , phytosterol oxides effects have been less extensively investigated . This study evaluates the production of inflammatory biomarkers ( IL-1β , P10145 , P22301 , TNFα ) and the influence of gene expression transporters and enzymes related to cholesterol absorption and metabolism ( Q9UHC9 , Q9H222 /8 , HMGCoA , ACAT ) produced by 7-ketosterols ( stigmasterol/cholesterol ) in Caco-2 cells . These effects were linked to intracellular signaling pathways by using several inhibitors . Results showed 7-ketostigmasterol to have a greater proinflammatory potential than 7-ketocholesterol . In non-pre-treated cells , only efflux transporters were down-regulated by 7-ketosterols , showing a greater influence upon Q9H222 expression . Cell-pre-incubation with bradykinin induced changes in ABCG expression levels after 7-ketostigmasterol-incubation ; however , the energetic metabolism inhibition reduced Q9UHC9 expression only in 7-ketocholesterol-incubated cells . In non-pre-treated cells , HMG- DB01992 was up-regulated by both 7-ketosterols . However , exposure to inhibitors down-regulated the expression levels , mainly in 7-ketocholesterol-incubated cells . While ACAT expression values in non-pre-treated cells were unchanged , exposure to inhibitors caused down-regulation of mRNA levels . These results suggest that internalization and excretion of 7-ketostigmasterol is probably influenced by [Ca]i , which also could mediate HMGCoA activity in POPs metabolism . However , energetic metabolism and reducing equivalents exert different influences upon the 7-ketosterol internalization . An absolute role of the PKC-dependent NF-kappaB activation for induction of P14780 in hepatocellular carcinoma cells . Matrix metalloproteinases ( MMPs ) play an important role in inflammation , tumor cell invasion , and metastasis . We found that phorbol-12-myristate-13-acetate ( PMA ) -stimulated invasion of the hepatocellular carcinoma ( HCC ) SNU-387 and SNU-398 cells and that PMA induced the secretion of P14780 in the cells , but did not induce the secretion of P08253 . The PMA-induced P14780 secretion was abolished by treatment of a pan-protein kinase C ( PKC ) inhibitor , GF109203X , and an inhibitor of NF-kappaB activation , sulfasalazine , and partly inhibited by treatment of inhibitors of P29323 pathway , PD98059 and U0126 . In addition , the PMA-stimulated activation of the P14780 promoter was completely inhibited by a mutation of the NF-kappaB site within the P14780 promoter , but not completely by mutations of two AP-1 sites . Moreover , the P14780 induction by P14210 and P01375 was also completely inhibited by GF109203X and sulfasalazine , but not by PD98059 and U0126 . These data demonstrate that the PKC-dependent NF-kappaB activation is absolute for P14780 induction and that the PKC-dependent P29323 activation devotes to increase the expression level of P14780 , in HCC cells . DB04540 metabolite , 5-cholesten-3β-25-diol-3-sulfate , promotes hepatic proliferation in mice . Oxysterols are well known as physiological ligands of liver X receptors ( LXRs ) . Oxysterols , 25-hydroxycholesterol ( 25HC ) and 27-hydroxycholesterol as endogenous ligands of LXRs , suppress cell proliferation via LXRs signaling pathway . Recent reports have shown that sulfated oxysterol , 5-cholesten-3β-25-diol-3-sulfate ( 25HC3S ) as LXRs antagonist , plays an opposite direction to oxysterols in lipid biosynthesis . The present report was to explore the effect and mechanism of 25HC3S on hepatic proliferation in vivo . Following administration , 25HC3S had a 48 h half life in the circulation and widely distributed in mouse tissues . Profiler™ PCR array and RTqPCR analysis showed that either exogenous or endogenous 25HC3S generated by overexpression of oxysterol sulfotransferase ( SULT2B1b ) plus administration of 25HC significantly up-regulated the proliferation gene expression of Wt1 , Pcna , cMyc , cyclin A , FoxM1b , and CDC25b in a dose-dependent manner in liver while substantially down-regulating the expression of cell cycle arrest gene Chek2 and apoptotic gene Apaf1 . Either exogenous or endogenous administration of 25HC3S significantly induced hepatic DNA replication as measured by immunostaining of the P12004 labeling index and was associated with reduction in expression of LXR response genes , such as O95477 and SREBP-1c . Synthetic LXR agonist T0901317 effectively blocked 25HC3S-induced hepatic proliferation . CONCLUSIONS : 25HC3S may be a potent regulator of hepatocyte proliferation and oxysterol sulfation may represent a novel regulatory pathway in liver proliferation via inactivating LXR signaling .

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Activation of Gi-coupled receptors releases a tonic state of inhibited platelet aggregation . Single-receptor pharmacology does not satisfactorily explain the physiology of the ADP-induced platelet aggregation response . It has been shown that , in addition to Gq-coupled receptor activation , one Gi-coupled receptor , either the ADP P2T or the alpha2-adrenoceptor , is required for elicitation of aggregation . The underlying mechanism of this action , however , has not been elucidated . By systematically assaying the entire time course of the aggregation and its fade using two methods of aggregometry , we have investigated the role of graded activation of these two Gi-coupled receptors . We demonstrate that constant activation of either of two Gq-coupled receptors , the ADP P47900 or the 5- Q13049 , and incremental activation of either of the two Gi-coupled receptors , tightly regulates the aggregation response in vitro , through the apparent release of a tonic inhibition of platelet aggregation . This tightly regulated release of inhibition , which appears analogous to the phenomena of disinhibition observed in the central nervous system , may be instrumental for the continuous adaptation of the aggregation response to variable physiological conditions . Serotonin skews human macrophage polarization through P41595 and P34969 . Besides its role as a neurotransmitter , serotonin ( 5-hydroxytryptamine , 5HT ) regulates inflammation and tissue repair via a set of receptors ( 5HT(1-7) ) whose pattern of expression varies among cell lineages . Considering the importance of macrophage polarization plasticity for inflammatory responses and tissue repair , we evaluated whether 5HT modulates human macrophage polarization . 5HT inhibited the LPS-induced release of proinflammatory cytokines without affecting P22301 production , upregulated the expression of M2 polarization-associated genes ( P05120 , P07996 , Q9NY15 , Q86Y22 ) , and reduced the expression of M1-associated genes ( P08476 , P41597 , P39900 , P05121 , P29016 , O94788 ) . Whereas only 5HT(7) mediated the inhibitory action of 5HT on the release of proinflammatory cytokines , both 5HT(2B) and 5HT(7) receptors mediated the pro-M2 skewing effect of 5HT . In fact , blockade of both receptors during in vitro monocyte-to-macrophage differentiation preferentially modulated the acquisition of M2 polarization markers . 5HT(2B) was found to be preferentially expressed by anti-inflammatory M2( P09603 ) macrophages and was detected in vivo in liver Kupffer cells and in tumor-associated macrophages . Therefore , 5HT modulates macrophage polarization and contributes to the maintenance of an anti-inflammatory state via 5HT(2B) and 5HT(7) , whose identification as functionally relevant markers for anti-inflammatory/homeostatic human M2 macrophages suggests their potential therapeutic value in inflammatory pathologies . Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors : importance of heparin cofactor II . DB01109 ( HEP ) prevents thrombus formation ( TF ) and thrombus growth ( TG ) , by accelerating thrombin ( THR ) inhibition by antithrombin III ( P01008 ) . Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II ( HCII ) , can inhibit TF and TG as effectively as HEP . This study compared the antithrombotic effects of HEP and another agent , DB06271 ( SLX ) which catalyzes thrombin inhibition by P01008 and HCII simultaneously . TF was induced in rabbit jugular veins , using the stasis/hypercoagulation model . TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins . HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg , respectively . SLX ( 16 anti-thrombin U/kg or 260 micrograms/kg ) was more effective than HEP ( 120 anti-thrombin U/kg or 800 micrograms/kg ) in preventing TG when administered either as a bolus or by continuous infusion . These data suggest that agents which accelerate THR inhibition by both P01008 and HCII simultaneously , can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP . [ Recent advances on the studies of the platelet 's inhibition and aggregation. State of the art of new Q9H244 antagonists ] . The interaction of ADP with its platelet receptor Q9H244 plays a crucial role in platelet activation and thrombogenesis . This article reviews the pharmacology and clinical trials of specific antagonists of Q9H244 . DB00758 is a thienopyridine with proven antithrombotic efficacy , but it has some important drawbacks : i ) it is a pro-drug that needs to be metabolized to its active metabolite ; ii ) it has a delayed onset and offset of action ; iii ) there is high inter-individual variability in pharmacological response . Prasugrel is also a thienopyridine , with faster onset of action and more uniform inhibition of platelet function compared to clopidogrel , accounting for lower incidence of ischemic events in patients with acute coronary syndromes ( ACS ) undergoing percutaneous coronary intervention ( P05154 ) and higher incidence of both non-CABG ( Coronary Artery Bypass Grafting ) related bleeding complications . Two direct and reversible Q9H244 antagonists , cangrelor and ticagrelor , are characterized by rapid onset and reversal of platelet inhibition . DB06441 did not prove superior to clopidogrel in preventing thrombotic events in patients undergoing P05154 . DB08816 proved to be superior to clopidogrel in preventing major adverse cardiac events in ACS patients , but was , like prasugrel , was associated with higher frequency of non-CABG-related bleeding complications . A shorter period of drug discontinuation before surgery was necessary in ticagrelor-treated patients compared to clopidogrel-treated patients to limit the severity of post-surgical bleeding . New Q9H244 blockers . A number of new antiplatelet agents currently in development are anticipated to improve clinical outcomes and safety benefits in patients with acute coronary syndrome ( ACS ) . This article reviews the pharmacology and clinical development of three of these agents : prasugrel , cangrelor , and ticagrelor . Prasugrel , a third-generation , oral thienopyridine , has been shown to be superior to clopidogrel , the current gold standard , in preventing ischemic events in patients with ACS undergoing percutaneous coronary intervention ( P05154 ) , although the bleeding rate was higher . DB06441 , a chemical analog of adenosine triphosphate , is a potent direct platelet Q9H244 antagonist . In development as an intravenous agent , cangrelor is currently being evaluated in two phase III studies in patients requiring P05154 . DB08816 is the first of a new class of orally available antiplatelet agents antagonizing the effects of ADP mediated by Q9H244 ; it is currently being studied in a phase III trial in patients with ACS . ( N ) -methanocarba-2MeSADP ( MRS2365 ) is a subtype-specific agonist that induces rapid desensitization of the P47900 receptor of human platelets . DB00640 diphosphate ( ADP ) initiates and maintains sustained aggregation of platelets through simultaneous activation of both the Gq-coupled P47900 receptor and the Gi-coupled Q9H244 receptor . We recently described the synthesis and P47900 receptor-specific agonist activity of ( N ) -methanocarba-2MeSADP ( MRS2365 ) . Consequences of selective activation of the P47900 receptor by MRS2365 have been further examined in human platelets . Whereas MRS2365 alone only induced shape change , addition of MRS2365 following epinephrine treatment , which activates the Gi/z-linked , alpha2A-adrenergic receptor , resulted in sustained aggregation that was indistinguishable from that observed with ADP . Conversely , the platelet shape change promoted by ADP in the presence of the P08514 /IIIa antagonist eptifibatide was similar to that promoted by MRS2365 . Preaddition of the high affinity P47900 receptor antagonist MRS2500 inhibited the effect of MRS2365 , whereas addition of MRS2500 subsequent to MRS2365 reversed the MRS2365-induced shape change . Preactivation of the P47900 receptor with MRS2365 for 2 min resulted in marked loss of capacity of ADP to induce aggregation as evidenced by a greater than 20-fold rightward shift in the concentration effect curve of ADP . This inhibitory effect of P47900 receptor activation was dependent on the concentration of MRS2365 ( EC50 = 34 nm ) . The inhibitory effect of preincubation with MRS2365 was circumvented by activation of the Gq-coupled 5- Q13049 receptor suggesting that MRS2365 induces loss of the ADP response as a consequence of desensitization of the Gq-coupled P47900 receptor . The time course of MRS2365-induced loss of aggregation response to epinephrine was similar to that observed with ADP . These results further demonstrate the P47900 receptor selectivity of MRS2365 and illustrate the occurrence of agonist-induced desensitization of the P47900 receptor of human platelets studied in the absence of Q9H244 receptor activation . (Pro)renin promotes fibrosis gene expression in P29320 cells through a Nox4-dependent mechanism . The (pro)renin receptor ( PRR ) has recently been demonstrated to bind equally well renin and its precursor , prorenin , leading to a similar intracellular signaling independent of angiotensin II . In this study , we report that human embryonic kidney cells ( P29320 ) exposed to renin or prorenin for 24 h in the presence of a blocking concentration of the angtiotensin-converting enzyme inhibitor perindoprilate increased superoxide anion production as measured by luminescence ( lucigenin ) and electron spin resonance spectroscopy ( hydroxylamine radical transition ) . Also , both renin and prorenin increased Nox4 expression while Nox2 , p47(phox) , and p67(phox) remained unchanged . In an investigation of the effects of renin and prorenin on fibrosis genes , it appeared that both proteins stimulated transforming growth factor-β ( TGF-β ) , fibronectin , and plasminogen activator inhibitor type 1 ( P05121 ) expression and therefore participated to an overall switch toward a profibrotic state of the kidney cells . When the cells were transfected with a siRNA targeting the PRR , Nox4 expression was efficiently prevented as well as the increase in superoxide production , TGF-β , fibronectin , and P05121 . Finally , we demonstrated that transfection of the cells with a Nox4-specific small interfering ( si ) RNA also prevented fibrosis gene expression following treatment with renin or prorenin . The results demonstrate that renin and prorenin , through their specific membrane receptor and independently of angiotensin II , promote fibrosis gene expression via a Nox4-dependent mechanism . Clinical effects and outcomes with new Q9H244 inhibitors in ACS . Thienopyridines have become the cornerstone of treatment for percutaneous coronary intervention although no survival benefit has ever been shown with clopidogrel despite increasing loading doses . Newly developed Q9H244 inhibitors are more potent , more predictable , and have a faster onset of action than clopidogrel , characteristics that make them particularly attractive for high-risk percutaneous coronary intervention ( P05154 ) . Four new Q9H244 inhibitors have been tested each of them having particular individual properties . Prasugrel is an oral pro-drug leading to irreversible blockade of the Q9H244 receptor and is approved worldwide for ACS P05154 . DB08816 is a direct-acting and reversible inhibitor of the Q9H244 receptor with potentially more pleiotropic effects . DB06441 is an intravenous direct and reversible inhibitor of the Q9H244 receptor providing the highest level of inhibition , and elinogrel is an intravenous and oral Q9H244 antagonist with a direct and reversible action . Both prasugrel and ticagrelor , opposed to clopidogrel , have shown that stronger Q9H244 inhibition led respectively to significant 19 and 16 % relative risk reduction of a similar primary end point combining cardiovascular death , non-fatal myocardial infarction , or non-fatal stroke . Both drugs showed a significant 0.6 % absolute excess of TIMI major bleeding not related to CABG surgery . Because in clinical trials , patients perceived to be at higher risk of bleeding usually are excluded , the risk of major and even fatal bleeding might even be higher in a ' real-world ' setting , i.e. in the elderly patient with comorbidities . On the other hand , these newly developed Q9H244 inhibitors decrease mortality after P05154 compared with clopidogrel . The risk/benefit ratio is particularly favorable in P05154 for patients with STEMI . Glycoprotein IIb/IIIa and Q9H244 receptor antagonists yield additive inhibition of platelet aggregation , granule secretion , soluble P29965 release and procoagulant responses . Glycoprotein IIb/IIIa ( P08514 /IIIa ) antagonists , including abciximab and tirofiban , are administered concurrently with clopidogrel , a Q9H244 antagonist , and aspirin in some patients undergoing percutaneous coronary intervention . We studied the effects of , and interactions between , abciximab , tirofiban , aspirin and the Q9H244 antagonist cangrelor on platelet aggregation , alpha and dense granule secretion and procoagulant responses in vitro . Blood was obtained from healthy volunteers . Platelet aggregation , dense granule secretion , alpha granule secretion ( P05121 and soluble P29965 levels ) and procoagulant responses ( annexin-V and microparticle formation ) were assessed using collagen and thrombin receptor activating peptide ( TRAP ) as agonists . All the antagonists used singularly inhibited collagen-induced responses . Combinations of abciximab or tirofiban with aspirin and/or cangrelor gave additive inhibition with the greatest effect seen when abciximab or tirofiban was combined with both aspirin and cangrelor . DB06441 inhibited TRAP-induced responses and , again , there was additive inhibition of these parameters when abciximab or tirofiban were combined with cangrelor . The P08514 /IIIa receptor plays an important role in amplification of platelet activation such that there are important interactions between P08514 /IIIa antagonists and inhibitors of both Q9H244 receptor activation and , to a lesser extent , thromboxane A2 generation . These interactions are likely to have important influences on the safety and efficacy of combination anti-platelet therapies . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . DB06441 for treatment during percutaneous coronary intervention . Dual antiplatelet therapy consisting of aspirin and a Q9H244 -receptor antagonist is important for preventing major adverse cardiovascular events in patients managed with percutaneous coronary intervention ( P05154 ) . The current Q9H244 -receptor antagonists are only available for oral administration and exhibit a delayed onset of action . Furthermore , several days are required for platelet function to return to normal following cessation of therapy . DB06441 is an intravenous DB00171 analog that directly , selectively and reversibly inhibits Q9H244 receptors on platelets . A 30-μg/kg bolus dose followed by a 4-μg/kg per minute continuous infusion of cangrelor achieves peak concentration and maximal platelet inhibition within minutes of administration . DB06441 also demonstrates a fast offset as normal platelet function is restored 1-2 h after cessation of the infusion . Three large , double-blind , randomized trials - CHAMPION PLATFORM , CHAMPION P05154 and CHAMPION PHOENIX - assessed the efficacy and safety of cangrelor compared with clopidogrel ( during or immediately after P05154 ) or placebo in the setting of P05154 . In the most recent CHAMPION PHOENIX trial , cangrelor was superior to clopidogrel for preventing adverse cardiovascular events with no significant increase in major bleeding . Based on the clinical trial results combined with unique properties such as intravenous administration and fast onset and offset , cangrelor may provide benefit in certain patients undergoing P05154 . DB06441 : a review on pharmacology and clinical trial development . Dual antiplatelet therapy with aspirin and an oral ADP Q9H244 receptor antagonist is the standard-of-care for the prevention of ischemic events in patients with acute coronary syndrome or undergoing percutaneous coronary intervention ( P05154 ) . However , currently available ADP Q9H244 receptor antagonists have several limitations , such as interindividual response variability , drug-drug interactions , slow onset/offset and only oral availability . DB06441 is a reversible , potent , intravenous , competitive inhibitor of the ADP Q9H244 receptor that rapidly achieves near complete and predictable platelet inhibition . Along with reversible binding to the receptor cangrelor also has a very short half-life ( 3-5 min ) , which in turn results in a rapid offset of action . These properties make cangrelor a promising drug for clinical use in patients undergoing P05154 or patients waiting for major surgery but still require antiplatelet protection . This manuscript provides an update of the current status of knowledge on cangrelor , focusing on its pharmacologic properties and clinical trial development , including the BRIDGE and CHAMPION-PHOENIX trials . B7h triggering inhibits umbilical vascular endothelial cell adhesiveness to tumor cell lines and polymorphonuclear cells . Vascular endothelial cells ( ECs ) are key players in leukocyte recruitment into tissues and metastatic dissemination of tumor cells . ECs express B7h , which is the ligand of the Q9Y6W8 T cell costimulatory molecule . The aim of this work was to assess the effect of B7h triggering by a soluble form of Q9Y6W8 ( Q9Y6W8 -Fc ) on the adhesion of colon carcinoma cell lines to HUVECs . We found that B7h triggering inhibited HUVEC adhesiveness to HT29 and DLD1 cells ( by 50 and 35 % , respectively ) but not to HCT116 cells . The effect was dependent on the Q9Y6W8 -Fc dose and was detectable as early as 30 min after treatment and was still present after 24 h . It was inhibited by soluble anti- Q9Y6W8 reagents ( mAb and B7h-Fc ) and silencing of B7h on HUVECs , and it was not displayed by an F119S mutated form of Q9Y6W8 -Fc that does not bind B7h . HUVEC treatment with Q9Y6W8 -Fc did not modulate expression of adhesion molecules and cytokines , but it substantially downmodulated P29323 phosphorylation induced by P16581 triggering or osteopontin , which may influence HUVEC adhesiveness . Moreover , HUVEC treatment with Q9Y6W8 -Fc also inhibited adhesion of polymorphonuclear cells and several tumor cell lines from different origins . Therefore , the B7h- Q9Y6W8 interaction may modulate spreading of cancer metastases and recruitment of polymorphonuclear cells in inflammatory sites , which opens a view on the use of Q9Y6W8 -Fc as an immunomodulatory drug . State of the art of new Q9H244 antagonists . The interaction of ADP with its platelet receptor Q9H244 plays a crucial role in platelet activation and thrombogenesis . This article reviews the pharmacology and clinical trials of specific antagonists of Q9H244 . DB00758 is a thienopyridine with proven antithrombotic efficacy , but it has some important drawbacks : ( a ) it is a pro-drug that needs to be metabolized to its active metabolite ; ( b ) it has a delayed onset and offset of action and ( c ) there is high inter-individual variability in pharmacological response . Prasugrel is also a thienopyridine , with faster onset of action and a more uniform inhibition of platelet function compared to clopidogrel , accounting for lower incidence of ischemic events in patients with acute coronary syndromes ( ACS ) undergoing percutaneous coronary intervention ( P05154 ) and higher incidence of both non-CABG-related bleeding complications . Two direct and reversible Q9H244 antagonists , DB06441 and ticagrelor , are characterized by rapid onset and reversal of platelet inhibition . DB06441 is not superior to clopidogrel in preventing thrombotic events in patients undergoing P05154 . DB08816 is superior to clopidogrel in preventing major adverse cardiac events in ACS patients , but , like prasugrel , is associated with a higher frequency of non-CABG-related bleeding complications . A shorter period of drug discontinuation before surgery is necessary in ticagrelor-treated patients compared to clopiodgrel-treated patients to limit the severity of post-surgical bleeding . Modeling of Q14654 and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 -sensitive potassium ( K+ DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 ) . The drugs and the compounds were docked to the DB00171 -dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME/Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule . Genetics in gestational diabetes mellitus : association with incidence , severity , pregnancy outcome and response to treatment . Constant advances in gene mapping technology have allowed research to focus from rare monogenic disorders on common complex diseases involving multiple susceptibility genes-environment interactions . Gestational diabetes mellitus ( GDM ) is a heterogeneous pathogenic condition affecting 2-5 % of all pregnant women during pregnancy . GDM is considered to result when genetic predisposition is triggered by increased insulin resistance during pregnancy leading to what seems to be one of the primary characteristics of GDM , the pancreatic b-cell impairment . Genetic predisposition to GDM has been suggested given the occurrence of the disease within family members . Furthermore , GDM is reported to be often present in women with maturity onset diabetes of the young ( MODY ) gene mutations . In addition , candidate susceptibility gene variants have been suggested to increase the risk of GDM . These genes include glucokinase ( GCK ) , HLA antigens , insulin receptor ( P06213 ) , insulin-like growth factor-2 ( P01344 ) , P41235 , insulin gene ( P01308 -VNTR ) , plasminogen activator inhibitor 1 ( P05121 ) , potassium inwardly rectifying channel subfamily J , member 11 ( Q14654 ) , hepatocyte nuclear factor-4a ( P41235 ) . Identification of the possible underlying genetic factors of GDM would eventually enrich our knowledge on the pathophysiologic mechanism of the disease and contribute to the individualization of both prevention and treatment of complications for the mother and fetus . However , so far , little is known about the genetic basis of GDM and its potential clinical significance . This review focuses on possible gestational diabetes mellitus susceptibility genes and their association with the disease incidence and severity as well as the pregnancy outcome and the response to treatment . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . Emerging antithrombotic drugs for acute coronary syndrome . INTRODUCTION : Acute coronary syndrome ( ACS ) encompasses acute myocardial infarction ( MI ) and unstable angina . Activation of platelets and coagulation cascade plays a central role in the development of ACS . Over the past decade , there have been substantial improvements in the strategies for secondary prevention of ACS , including the development of more potent oral antiplatelet agents such as prasugrel and ticagrelor . However , therapies with even better efficacy and safety profiles and more rapid onset and offset of action would be desirable . AREAS COVERED : This review discusses the advantages and disadvantages of the currently available antithrombotic agents and describes the findings from recent clinical trials of three novel agents ; cangrelor ( an intravenous Q9H244 receptor antagonist ) , vorapaxar ( protease-activated receptor-1 inhibitor ) and rivaroxaban ( an oral factor Xa inhibitor ) . EXPERT OPINION : DB06441 appears more promising than clopidogrel when a very rapid onset and reversal of antiplatelet effect is needed . DB09030 in addition to standard oral antiplatelet therapy was effective in patients with prior MI , but was not safe in patients with a prior stroke . Low dose rivaroxaban decreased cardiovascular events and mortality in patients post-ACS compared to placebo , although bleeding was increased . DB06441 in percutaneous coronary intervention . DB06441 is a novel , intravenous Q9H244 receptor antagonist in development for use in percutaneous coronary intervention . Currently in Phase III testing , the reversible platelet inhibitor provides several inherent advantages over other Q9H244 receptor antagonists in this setting for the prevention of adverse cardiac events . Unlike the class of thienopyridines ( ticlopidine , clopidogrel and potentially soon to be available , prasugrel ) , cangrelor has nearly immediate onset after a bolus dose and a short half-life , and achieves maximal inhibition of ADP-mediated platelet function . DB06441 's distinct mechanism of action allows for intravenous administration and avoids both hepatic and renal metabolism . These unique characteristics make cangrelor a promising agent for use in cardiovascular patients undergoing percutaneous coronary intervention . DB06441 : a novel Q9H244 receptor antagonist . Antiplatelet therapy is critical in the prevention of thrombotic complications of acute coronary syndrome and percutaneous coronary interventions . Current antiplatelet agents ( aspirin , clopidogrel and glycoprotein IIb/IIIa antagonists ) have demonstrated the capacity to reduce major adverse cardiac events . However , these agents have limitations that compromise their clinical utility . The platelet Q9H244 receptor plays a central role in platelet function and is a focus in the development of antiplatelet therapies . DB06441 is a potent , competitive inhibitor of the Q9H244 receptor that is administered by intravenous infusion and rapidly achieves near complete inhibition of ADP-induced platelet aggregation . This investigational drug has been studied for use during coronary procedures and the management of patients experiencing acute coronary syndrome and is undergoing evaluation for use in the prevention of perioperative stent thrombosis . Tandospirone activates neuroendocrine and P29323 ( Q96HU1 kinase ) signaling pathways specifically through P08908 receptor mechanisms in vivo . Tandospirone , an azapirone , is a selective serotonin(1A) ( 5-HT(1A) ) receptor agonist . The effects of tandospirone on plasma hormones and on mitogen-activated protein ( Q96HU1 ) kinase activity in the brain of male rats were studied . Tandospirone produced a time- and dose-dependent increase in plasma levels of oxytocin , adrenocorticotropin ( DB01285 ) , corticosterone , and prolactin . The minimal dose of tandospirone that led to a significant elevation of plasma oxytocin , DB01285 , and prolactin levels was 1.0 mg/kg ( s.c. ) , while the minimal dose for corticosterone release was 3.0 mg/kg ( s.c. ) . The ED(50) of tandospirone was 1.3 mg/kg for oxytocin , 1.2 mg/kg for DB01285 , 3.0 mg/kg for corticosterone , and 0.24 mg/kg for prolactin . Pretreatment with the specific 5-HT(1A) receptor antagonist WAY 100,635 ( 0.3 mg/kg , s.c. ) completely blocked the effects of tandospirone on plasma levels of oxytocin , DB01285 , and corticosterone but shifted the dose-response curve for prolactin to the right . Tandospirone injection ( 10 mg/kg , s.c. ) stimulated the Q96HU1 kinase signaling cascade , specifically the phosphorylation of Q8NFH3 /44 extracellular signal-regulated kinase ( P29323 ) . Western blot analysis revealed a significant increase in phosphorylated P29323 ( p- P29323 ) levels in the hypothalamic paraventricular nucleus ( PVN ) as well as the dorsal raphe nucleus 5 min following tandospirone injection . These increases were blocked by pretreatment with WAY 100,635 ( 0.3 mg/kg ) . The results are the first evidence that systemic 5-HT(1A) receptor agonist administration produces a rapid increase in p- P29323 levels in vivo , providing further insight into the signaling mechanisms of the 5-HT(1A) receptor . Aripiprazole : a novel atypical antipsychotic drug with a uniquely robust pharmacology . Aripiprazole ( DB01238 ) is an atypical antipsychotic drug that has been recently introduced for clinical use in the treatment of schizophrenia . Aripiprazole has a unique pharmacologic profile that includes partial agonism at several G-protein coupled receptors ( GPCRs ) [ especially dopamine ( D2 ) and P08908 ] and antagonistic action at others ( especially 5- Q13049 ) . Clinical trials indicate that aripiprazole is effective in treating the positive and negative symptoms of schizophrenia . In short-term studies rapid onset of action ( within one week ) has been demonstrated . Preliminary data indicate that aripiprazole may also be effective in the treatment of manic symptoms of bipolar disorder . At recommended doses , aripiprazole appears to be safe and well tolerated in most adult patients with schizophrenia and schizoaffective disorder . There is only limited information available on the use of aripiprazole in children and adolescents , and pilot data suggest that a revised dosing strategy , based on weight , is indicated in this population . In the long-term studies , the use of aripiprazole was associated with continued efficacy , good compliance and increased time-to-relapse . Aripiprazole represents the first functionally selective atypical antipsychotic drug . The influence of variation in the Q9H244 receptor gene on in vitro platelet inhibition with the direct Q9H244 antagonist cangrelor . Novel Q9H244 inhibitors are in development to overcome the occurrence of atherothrombotic events associated with poor responsiveness to the widely used Q9H244 inhibitor clopidogrel . DB06441 is an intravenously administered Q9H244 inhibitor that does not need metabolic conversion to an active metabolite for its antiplatelet action , and as a consequence exhibits a more potent and consistent antiplatelet profile as compared to clopidogrel . It was the objective of this study to determine the contribution of variation in the Q9H244 receptor gene to platelet aggregation after in vitro partial Q9H244 receptor blockade with the direct antagonist cangrelor . Optical aggregometry was performed at baseline and after in vitro addition of 0.05 and 0.25 microM cangrelor to the platelet-rich plasma of 254 healthy subjects . Five haplotype-tagging (ht)-SNPs covering the entire Q9H244 receptor gene were genotyped ( rs6798347C > t , rs6787801T > c , rs9859552C > a , rs6801273A > g and rs2046934T > c [ T744C ] ) and haplotypes were inferred . The minor c allele of SNP rs6787801 was associated with a 5 % lower 20 microM ADP-induced peak platelet aggregation ( 0.05 microM cangrelor , p < 0.05 ) . Aa homozygotes for SNP rs9859552 showed 20 % and 17 % less inhibition of platelet aggregation with cangrelor when compared to CC homozygotes ( 0.05 and 0.25 microM cangrelor respectively ; p < 0.05 ) . Results of the haplotype analyses were consistent with those of the single SNPs . Polymorphisms of the Q9H244 receptor gene contribute significantly to the interindividual variability in platelet inhibition after partial in vitro blockade with the Q9H244 antagonist cangrelor . Analysis of breast cancer related gene expression using natural splines and the Cox proportional hazard model to identify prognostic associations . Many studies correlating gene expression data to clinical parameters assume a linear increase or decrease of the clinical parameter under investigation with the expression of a gene . We have studied genes encoding important breast cancer-related proteins using a model for survival-type data that is based on natural splines and the Cox proportional hazard model , thereby removing the linearity assumption . Expression data of 16 genes were studied in relation to metastasis-free probability in a cohort of 295 consecutive breast cancer patients treated at The Netherlands Cancer Institute . The independent predictive power for disease outcome of the 16 individual genes was tested in a multivariable model with known clinical and pathological risk factors . There is a linear relationship between increasing expression and a higher or lower hazard for distant metastasis for P03372 , Q15303 , P15692 , O96020 , Q15910 , and Q96NZ9 ; for P04626 , P21860 , P24385 , P24864 , O75530 , P61073 , P32248 , P48061 , and P05121 there is no clear increase or decrease ; and for P00533 there seems to be a non-linear relation . Multivariable analysis showed that the 70-gene prognosis profile outperforms all the other variables in the model ( hazard-rate 5.4 , 95 % CI 2.5-11.7 ; P = 0.000018 ) . P00533 -expression seems to have a non-linear relation with disease outcome , indicating that lower but also higher expression of P00533 are associated with worse outcome compared to intermediate expression levels ; the other genes show no or a linear relation . A critical appraisal of the functional evolution of Q9H244 antagonists as antiplatelet drugs . Q9H244 receptor mediated inhibition of platelet aggregation is one of the most explored and exploited pathways in antiplatelet drug therapy to prevent ischemic events in patients undergoing percutaneous coronary intervention ( P05154 ) for the treatment of the acute coronary syndrome ( ACS ) . DB00208 , DB00758 , Prasugrel , DB08816 , DB06441 and Elinogrel are the Q9H244 inhibitors that act as antiplatelet drugs . In this review , the features of these drugs and the factors reported to be responsible for drug resistance or drug ineffectiveness were described . The features like drug metabolism , reversible or irreversible binding of drugs to their target protein and the mode of administration were observed to evolve along with the antiplatelet drugs . These features also include the drug-drug interactions , the pharmacogenetics and pharmacodynamics of Q9H244 inhibitors . We attempted to critically analyze how the desirable features were met by the Q9H244 inhibitors in the course of time . This review provides an overview of the evolution of Q9H244 inhibitors and may guide the researchers to develop better antiplatelet drugs in the future . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Oxidized DB00171 ( oATP ) attenuates proinflammatory signaling via P2 receptor-independent mechanisms . Periodate-oxidized DB00171 ( oATP ) , which covalently modifies nucleotide-binding proteins , can significantly attenuate proinflammatory signaling . Although the Q99572 nucleotide receptor ( P2X7R ) is irreversibly antagonized by oATP , it is unclear whether anti-inflammatory actions of oATP are predominantly mediated via its actions on P2X7R . Here , we describe inhibitory effects of oATP on proinflammatory responses in three human cell types that lack expression of P2X7R : human umbilical vein endothelial cells ( HUVEC ) , HEK293 cells , and 1321N1 astrocytes . oATP decreased by 40-70 % the secretion of interleukin ( IL ) -8 stimulated by tumor necrosis factor-alpha ( P01375 ) in all three cell types , by IL-1beta in HUVEC and 1321N1 cells , and by endotoxin in HUVEC . Attenuation of P01375 -stimulated P10145 secretion by oATP was similar in wild-type P29320 cells or P29320 cells stably expressing recombinant P2X7R . oATP also attenuated cytokine-stimulated expression of nuclear factor-kappaB-luciferase reporter genes expressed in P29320 or 1321N1 cells , but did not affect the rapid downregulation of IkappaB. oATP had no effect on uridine triphosphate-induced activation of native P41231 receptors in P29320 cells , but reduced the potency and efficacy of ADP as an agonist of native P47900 receptors . However , inhibition of P47900 receptors with the specific antagonist MRS2216 did not mimic the effects of oATP on P01375 -stimulated P10145 secretion . Although 1321N1 astrocytes lack expression of any known P2 receptor subtypes , oATP markedly inhibited ecto-ATPase activity in these cells , resulting in a significant accumulation of extracellular DB00171 . In summary , oATP can attenuate proinflammatory signaling by mechanisms independent of the expression or activation of known P2 receptor subtypes . Chronic daily tadalafil prevents the corporal fibrosis and veno-occlusive dysfunction that occurs after cavernosal nerve resection . OBJECTIVES : To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 ( O76074 ) inhibitor , tadalafil , has a similar effect to that of the shorter half-life O76074 inhibitors sildenafil and vardenafil , and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction ( CVOD ) occurring after cavernosal nerve ( CN ) injury . MATERIALS AND METHODS : Male rats ( 10 per group ) had either a sham operation , unilateral CN resection ( P21554 ) or bilateral P21554 , and were left untreated or given retrolingually 5 mg/kg per day of tadalafil . After 45 days , CVOD was assessed via cavernosometry , and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry ( followed by quantitative image analysis ) , Western blots , and ad hoc methods . RESULTS : DB00820 treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after P21554 compared with sham-operated rats . DB00820 also normalized the increase in penile shaft collagen content , and the reduction in corporal smooth muscle cell ( SMC ) content , SMC/collagen , and replication index , and improved the lower collagen III/I ratio and the increase in apoptotic index , caused by P21554 , compared with sham operation . There were no effects of tadalafil on increased transforming growth factor beta1 , inducible nitric oxide synthase and xanthine oxidoreductase levels . CONCLUSIONS : A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage , as effectively as the previously reported continuous treatment with vardenafil or sildenafil , through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction . Transitioning patients from cangrelor to clopidogrel : pharmacodynamic evidence of a competitive effect . BACKGROUND : DB06441 is a direct , parenteral , and reversible inhibitor of the platelet Q9H244 receptor currently undergoing Phase III testing . As many individuals treated acutely with cangrelor will often be treated long-term with a thienopyridine , it is important to determine the effects of concurrent cangrelor and clopidogrel administration . METHODS AND RESULTS : Ten healthy volunteers received a 600 mg oral loading dose of clopidogrel and then underwent serial platelet function monitoring for 6 h . Two weeks later these same individuals received a 600 mg clopidogrel loading dose simultaneously with a cangrelor IV bolus ( 30 microg/kg ) and a 2-hour infusion ( 4 microg/kg/min ) . A separate group of ten volunteers received a 600 mg clopidogrel loading dose after administration of a cangrelor bolus and a 1-hour infusion . The effects on ADP-induced platelet activation and aggregation were evaluated by flow cytometry , whole-blood electrical impedance , and light-transmittance aggregometry . DB06441 and clopidogrel alone achieved the expected levels of platelet inhibition . However , the sustained platelet inhibition anticipated for clopidogrel treatment did not occur when cangrelor was initiated simultaneously . No such effect was found when clopidogrel was started upon completion of the cangrelor infusion . CONCLUSION : To achieve sustained platelet Q9H244 inhibition in patients treated with cangrelor , clopidogrel administration should be started when the cangrelor infusion is terminated . A field synopsis and meta-analysis of genetic association studies in peripheral arterial disease : The CUMAGAS-PAD database . In an electronic search of the literature , the authors systematically retrieved all published studies that investigated genetic susceptibility to peripheral arterial disease ( PAD ) . They created a comprehensive database of all eligible studies , collecting detailed genetic and bioinformatics data on each polymorphism . Data from eligible studies were synthesized using meta-analysis techniques . Gene variants were classified into distinct pathophysiologic pathways , and their potential involvement in PAD pathogenesis was determined . Forty-one publications that examined 44 gene polymorphisms were included . For 37 polymorphisms , the variant form had a functional effect . Twenty-three polymorphisms in 22 potential PAD candidate genes ( F2 , P02675 , P42898 , P05106 , P12821 , AGT , P05231 , P13500 , P05362 , P16581 , P14780 , P37231 , P03956 , P35611 , Q9H244 , P11150 , Q13093 , Q8WTV0 , P08254 , P55157 , P08519 , P32297 ) showed a significant association in individual studies . Eighty-eight percent of the studies had statistical power of less than 50 % , and in 15 studies the genotype distribution in the control group did not conform to Hardy-Weinberg equilibrium . Data on 12 polymorphisms ( P12259 1691 G/A , P42898 677C/T , F2 20210 G/A , P05106 1565 T/C , P12821 I/D , AGT 704C/T , AGT -6G/A , AGT 733C/T , P05231 -174 G/C , P14780 -1562C/T , P05362 1462A/G , P32297 831C/T ) were synthesized , and a positive association was found for 3 ( P05231 -174 G/C , P05362 1462A/G , P32297 831C/T ) . Serotonin induces the expression of tissue factor and plasminogen activator inhibitor-1 in cultured rat aortic endothelial cells . Serotonin ( 5-hydroxytryptamine , or 5-HT ) , released from activated platelets , not only accelerates aggregation of platelets but also is known to promote mitosis , migration , and contraction of vascular smooth muscle cells ( VSMCs ) . These effects are considered to contribute to thrombus formation and atherosclerosis . The aim of this study was to investigate the effects of 5-HT on the expressions of coagulative and fibrinolytic factors in rat aortic endothelial cells . Endothelial cells were stimulated with various concentrations of 5-HT ( 0.1 approximately 10 microM ) , and the expressions of tissue factor ( TF ) , tissue factor pathway inhibitor ( P10646 ) , plasminogen activator inhibitor-1 ( P05121 ) , and tissue-type plasminogen activator ( TPA ) messenger RNAs ( mRNAs ) were evaluated by Northern blot analysis . The activities of TF and P05121 were also measured . TF and P05121 mRNA were increased significantly in a concentration- and time-dependent manner . However , P10646 and TPA mRNA expression did not change . The inductions of TF and P05121 mRNAs were inhibited by a 5-HT1/5-HT2 receptor antagonist ( methiothepin ) and a selective 5- Q13049 receptor antagonist ( D6RGH6 -9042 ) . These results indicate that 5-HT increases procoagulant activity and reduces fibrinolytic activities of endothelial cells through the 5- Q13049 receptor . It was concluded that the modulation of procoagulant and hypofibrinolytic activities of endothelial cells by 5-HT synergistically promotes thrombus formation at the site of vessel injury with the platelet aggregation , VSMC contraction , and VSMC proliferation . Spin trapping agent phenyl-N-tert-butylnitrone prevents diisopropylphosphorofluoridate-induced excitotoxicity in skeletal muscle of the rat . Indirect evidence suggests that reactive oxygen species ( ROS ) may mediate muscle fiber necrosis following muscle hyperactivity induced by the anticholinesterase diisopropylphosphorofluoridate ( DB00677 ) . Pronounced muscle fasciculations and muscle fiber necrosis were seen when acetylcholinesterase ( P22303 ) activity was reduced to less than 30 % of control . The spin trapping agent phenyl-N-tert-butylnitrone ( PBN ) was used in vivo to directly assess the formation of ROS during DB00677 ( 1.75 mg/kg , s.c. ) induced muscle hyperactivity . Pretreatment with PBN ( 300 mg/kg , i.p. ) , the concentration necessary for in vivo spin trapping , prevented muscle hyperactivity as well as necrosis and attenuated the DB00677 induced P22303 inhibition otherwise seen in DB00677 only treated rats . PBN had no effect when given after fasciculations were established . Muscle extracts from PBN and DB00677 treated rats subjected to electron spin resonance ( P03372 ) spectroscopy tested negative for ROS . While the role of PBN as an antioxidant is well established , its prophylactic effect against excitotoxity induced by an P22303 inhibitor are due to its protection of P22303 , an unexpected non-antioxidant action . The presence and function of dopamine type 2 receptors in boar sperm : a possible role for dopamine in viability , capacitation , and modulation of sperm motility . Several studies have shown that dopamine and other catecholamines are present in oviduct luminal fluid . We recently reported that dopamine type 2 receptors ( P14416 ) are present in a wide range of mammalian sperm , suggesting a role for dopaminergic signaling in events such as fertilization , capacitation , and sperm motility . In the present study , we used Western blot analysis to show that boar sperm express P14416 and that their activation with dopamine ( 100 nM ) has a positive effect on cell viability that can be correlated with AKT/ P31749 phosphorylation . DB01200 ( 100 nM ) and dopamine ( 100 nM and 10 muM ) increased tyrosine phosphorylation during the capacitation period . Immunofluorescence analysis indicated that P14416 localization is dynamic and depends on the capacitation stage , colocalizing with tyrosine phosphorylated proteins in the acrosome and midpiece region of capacitated boar sperm . This association was confirmed by coimmunoprecipitation analysis . We also showed that bromocriptine ( 100 nM ) and low-concentration dopamine ( 100 nM and 10 muM ) increased total and progressive motility of sperm . However , high concentrations of dopamine ( 1 mM ) decreased tyrosine phosphorylation and motility in in vitro sperm capacitation assays . This can be explained by the presence of the dopamine transporters ( Q01959 , official symbol Q01959 ) in sperm , as demonstrated by Western blot analysis and immunocytochemistry . Taken together , our results support the idea that dopamine may have a fundamental role during sperm capacitation and motility in situ in the female upper reproductive tract . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . DB06441 for treatment of coronary thrombosis . OBJECTIVE : To review and assess available literature on the chemistry , pharmacology , pharmacodynamics , pharmacokinetics , clinical studies , adverse events , drug interactions , special populations , and dosing and administration for cangrelor , a product in late stage Phase II clinical trials . DATA SOURCES : A literature search of MEDLINE ( 1966-March 2006 ) , International Pharmaceutical Abstracts ( 1970-February 2006 ) , and Cochrane database ( first quarter 2006 ) was conducted using key terms of cangrelor , AR-C69931MX , and Q9H244 receptor antagonist . Bibliographies of relevant articles were reviewed for additional references . The Medicines Company Web site was reviewed , and a company representative was contacted . STUDY SELECTION AND DATA EXTRACTION : Available English-language literature , including abstracts , preclinical studies , clinical trials , and review articles , was reviewed . DATA SYNTHESIS : DB06441 is a Q9H244 antagonist under development for treatment of acute coronary syndrome . DB06441 has been studied as an intravenous infusion in doses of 2 or 4 microg/kg/min . It inhibits platelet aggregation with rapid onset and offset and does not require metabolism for therapeutic activity . Published Phase II trials have demonstrated safety and inhibition of platelet aggregation . CONCLUSIONS : DB06441 is a promising investigational medication for inhibition of platelet aggregation in acute arterial coronary events . Phase II trials have shown safety and a greater inhibition of platelet aggregation over clopidogrel . Phase III trials will provide more definitive information on clinical efficacy and safety . Until then , the role of cangrelor is uncertain . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Nerve agent exposure elicits site-specific changes in protein phosphorylation in mouse brain . Organophosphorus ( OP ) compounds cause toxic symptoms , including convulsions , coma , and death , as the result of irreversible inhibition of acetylcholinesterase ( P22303 ) . The development of effective treatments to block these effects and attenuate long-term cognitive and motor disabilities that result from OP intoxication is hampered by a limited understanding of the CNS pathways responsible for these actions . We employed a candidate method ( called CNSProfile ) to identify changes in the phosphorylation state of key neuronal phosphoproteins evoked by the OP compound , diisopropyl fluorophosphate ( DB00677 ) . Focused microwave fixation was used to preserve the phosphorylation state of phosphoproteins in brains of DB00677 -treated mice ; hippocampus and striatum were analyzed by immunoblotting with a panel of phospho-specific antibodies . DB00677 exposure elicited comparable effects on phosphorylation of brain phosphoproteins in both C57BL/6 and FVB mice . DB00677 treatment significantly altered phosphorylation at regulatory residues on glutamate receptors , including Serine897 ( S897 ) of the Q9UHB4 DB01221 receptor . Q9UHB4 phosphorylation was bi-directionally regulated after DB00677 in striatum versus hippocampus . Q9UHB4 phosphorylation was reduced in striatum , but elevated in hippocampus , compared with controls . Q9UD71 phosphorylation in striatum was selectively increased at the Cdk5 kinase substrate , Threonine75 ( T75 ) . Phencynonate hydrochloride , a muscarinic cholinergic antagonist , prevented seizure-like behaviors and the observed changes in phosphorylation induced by DB00677 . The data reveal region-specific effects of nerve agent exposure on intracellular signaling pathways that correlate with seizure-like behavior and which are reversed by the muscarinic receptor blockade . This approach identifies specific targets for nerve agents , including substrates for Cdk5 kinase , which may be the basis for new anti-convulsant therapies . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . Autocrine regulation of γ-irradiation-induced DNA damage response via extracellular nucleotides-mediated activation of Q15077 and Q9H244 receptors . A key component of the response to DNA damage caused by ionizing radiation is DNA repair . Release of extracellular nucleotides , such as DB00171 , from cells plays a role in signaling via P2 receptors . We show here that release of DB00171 , followed by activation of P2Y receptors , is involved in the response to γ-irradiation-induced DNA damage . Formation of phosphorylated histone variant P16104 ( γ P16104 ) foci , which are induced in nuclei by DNA damage and contribute to accumulation of DNA-repair factors , was increased at 1-3h after γ-ray irradiation ( 2.0Gy ) of human lung cancer A549 cells . Focus formation was suppressed by pre-treatment with the ecto-nucleotidase apyrase . Pre-treatment with ecto-nucleotidase inhibitor ARL67156 or post-treatment with DB00171 or UTP facilitated induction of γ P16104 , indicating that extracellular nucleotides play a role in induction of γ P16104 foci . Next , we examined the effect of P2 receptor inhibitors on activation of ataxia telangiectasia mutated ( Q13315 ; a protein kinase ) and accumulation of Q12888 ( a DNA repair factor ) , both of which are important for DNA repair , at DNA damage sites . Q15077 receptor antagonist MRS2578 , Q9H244 receptor antagonist clopidogrel , and Q99572 receptor antagonists A438079 and oxATP significantly inhibited these processes . Release of DB00171 was detected within 2.5min after irradiation , but was blocked by A438079 . Activation of Q13315 and accumulation of Q12888 were decreased in Q15077 or Q9H244 receptor-knockdown cells . We conclude that autocrine/paracrine signaling through Q99572 -dependent DB00171 release and activation of Q15077 and Q9H244 receptors serves to amplify the cellular response to DNA damage caused by γ-irradiation . Pharmacokinetics and pharmacodynamics of a bolus and infusion of cangrelor : a direct , parenteral Q9H244 receptor antagonist . The purpose of this study is to evaluate the safety , tolerability , pharmacokinetics , and pharmacodynamics of cangrelor administered as an intravenous bolus plus a continuous infusion in healthy volunteers . Twenty-two healthy volunteers are randomized to receive 1 of 2 intravenous cangrelor dosing regimens : a 15-microg/kg bolus followed by a 2-microg/kg/min infusion or a 30-microg/kg bolus followed by a 4-microg/kg/min infusion . The infusion is continued for 60 minutes , and serial blood samples are obtained for evaluation of pharmacokinetic and pharmacodynamic parameters . Administration of an intravenous bolus followed by a continuous infusion rapidly achieves maximum concentrations of cangrelor that are associated with extensive platelet inhibition within 2 minutes . Moreover , extensive platelet inhibition is maintained throughout the infusion period with near-full recovery of platelet function within 60 to 90 minutes of terminating the infusion . The effect of high-dose cangrelor is more consistent and demonstrates a greater level of inhibition on adenosine diphosphate-induced P16109 expression ; how ever , no significant differences are observed between the 2 dosing regimens with regard to platelet aggregation or time to recovery of platelet function . DB06441 administered as an intravenous bolus followed by a continuous infusion in healthy volunteers offers rapid and reversible inhibition of platelet function . Normal and perturbed endothelial cells from canine femoral arteries and femoral veins exhibit heterogeneity in hemostatic properties and growth characteristics . BACKGROUND : We sought to examine the heterogeneity of endothelial cells from the same anatomic site but different vascular systems and described P04275 ( P04275 ) release and morphological change in response to injury-associated factor in femoral vessels from canine in vitro . METHODS : Levels of hemostatic factors ( P04275 , plasminogen activator inhibitor type 1( P05121 ) , antithrombin III ( P01008 ) , in tissue sections and cultured endothelial cells of canine femoral arteries and canine femoral veins were compared by the immunohistochemistry technique . In addition to comparing cell growth density and cell protein contents , cultured femoral arterial endothelial cells ( FAECs ) and cultured femoral venous endothelial cells ( FVECs ) were incubated with a series concentration of basic fibroblast factor ( P09038 ) ( 1 , 10 , 100 ng/ml ) for up to 48 hours to test the amount of P04275 secretion and morphological change . RESULTS : Both in tissue sections and cultured cells , the levels of P04275 are higher in FVECs than in FAECs . We were unable to differentiate the level of P05121 and P01008 difference between FAECs and FVECs. P09038 ( 10 ng/ml ) significantly increased P04275 secretion from cultured FAECs but not from FVECs . The size of cultured FAECs is smaller than of FVECs ; however , FAECs have higher amounts of protein contents than FVECs . CONCLUSIONS : These comparative studies provide evidence indicating that the characteristics of FVECs differ from those of FAECs . These differences may be indicated heterogeneity with either inherited or acquired thrombotic disease . Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 ) subunits of K( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 varies between tissues : Q09428 in beta-cells , SUR2A in cardiac muscle , and SUR2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K( DB00171 ) channel subunits . While K( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR2-type K( DB00171 ) channels ( e.g. , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K( DB00171 ) channels are to be avoided . Echicetin coated polystyrene beads : a novel tool to investigate GPIb-specific platelet activation and aggregation. P04275 /ristocetin ( P04275 /R ) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin , which also binds P04275 . These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets . Here , we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads , which specifically activate GPIb . We compared platelet activation induced by echicetin beads to P04275 /R . Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling . Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets , while under the same conditions P04275 /R treatment led only to αIIbβ3-independent platelet agglutination . The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation , while the total amount of echicetin used for activation is not critical . Echicetin beads induced strong phosphorylation of several proteins including p38 , P29323 and P31749 . Synergistic signaling via Q9H244 and thromboxane receptor through secreted ADP and TxA2 , respectively , were important for echicetin bead triggered platelet activation . Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation . Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways . P2Y receptor antagonists in thrombosis . The dual role of P47900 and Q9H244 receptors in platelet aggregation by ADP has been firmly established , based on the action of selective inhibitors , gene targeting in mice and human genetic evidence . Both of these receptor subtypes constitute targets for antithrombotic agents , and compounds with a dual action might also be of interest . However , the agents currently on the market ( ticlopidine and clopidogrel ) , or known to be in development ( cangrelor , DB08816 and prasugrel ) , all target the Q9H244 receptor . The thienopyridines ( ticlopidine , clopidogrel and prasugrel ) irreversibly inactivate the Q9H244 receptor via the covalent binding of an active metabolite generated in the liver , while the other compounds are competitive antagonists . DB06441 , an DB00171 derivative , is suitable for intravenous perfusion , whereas DB08816 is in clinical development as an orally active agent . Agonism at P41595 receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5-hydroxytryptamine2B ( P41595 ) receptors . To evaluate whether agonism at P41595 receptors is a phenomenon of the class of the ergolines , we studied P41595 receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC50 8.42 and 8.72 ) . DB01200 acted as a partial agonist ( pEC50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5-HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 receptors seems not to be a class effect of the ergolines . Q9H244 inhibitors : pharmacologic mechanism and clinical relevance . Platelets play a critical role in the pathogenesis of atherothrombotic processes and inhibition of platelet aggregation by antiplatelet therapy is essential and really important in the acute coronary syndromes or in the setting of percutaneous coronary intervention . The first family of adenosine diphosphate Q9H244 receptors inhibiting drug is represented by thienopyridines and among these ticlopidine was the first approved by Food and Drug Administration ; actually its use is discouraged because of its potential side effects ( neutropenia , anemia , gastrointestinal distress and thrombotic thrombocytopenic purpura ) . The second generation of thienopyridines is represented by clopidogrel that has replaced ticlopidine in the clinical practice ; clopidogrel has the largest clinical experience . Prasugrel represents the third generation . It inhibits platelet aggregation by irreversibly blocking the adenosine diphosphate Q9H244 receptor . DB08816 , DB06441 and Enilogrel represent the last generation of thienopyridines . This review is focused on the effects of adenosine diphosphate Q9H244 inhibitors . Gating properties of Q14524 mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 mutations in 5 symptomatic LQT3 patients with different responses to Mex ( 6 to 8 mg . kg(-1) . d(-1) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; >/= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na(+) current from P29320 293 cells transfected with wild-type ( WT ) or mutant Nav1.5 . All mutations showed impaired inactivation of Na(+) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V(1/2) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P=P1332L > S941N=WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V(1/2) of steady-state inactivation correlate with the clinical response observed in LQT3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT3 . New frontiers in the management of acute coronary syndromes : cangrelor and elinogrel . The activation and aggregation of platelets at sites of vascular injury or near to implanted stent are pivotal in the development of thrombotic events during and after an acute coronary syndrome ( ACS ) or a percutaneous coronary intervention ( P05154 ) . For that reason , an exclusively oral dual antiplatelet treatment regimen with platelet Q9H244 receptor antagonists in addition to the cyclooxygenase inhibitor aspirin has become the cornerstone of treatment in that contest . However , every trial underlines the same problem : if maximizing antiplatelet therapy significantly attenuates ischemic events in patients with coronary artery disease , on the other side it may also increase bleeding phenomena . These limitations have prompted a search for novel antiplatelet agents with a more favorable risk-benefit ratio . Moreover , an early onset of action is desirable during P05154 and an early offset after bleeding events . Two novel antiplatelet agents , DB06441 and Elinogrel , are available in intravenous form ( Elinogrel also in oral form ) and expand this context . Recent trials have tested them against DB00758 regarding efficacy and safety outcomes.This review aimed at providing an overview on intravenous emerging compounds and recent patents in the setting of ACS and P05154 . Alteration of synaptic activity-regulating genes underlying functional improvement by long-term exposure to an enriched environment in the adult brain . BACKGROUND : Housing animals in an enriched environment ( EE ) enhances behavioral function . However , the mechanism underlying this EE-mediated functional improvement and the resultant changes in gene expression have yet to be elucidated . OBJECTIVES : We attempted to investigate the underlying mechanisms associated with long-term exposure to an EE by evaluating gene expression patterns . METHODS : We housed 6-week-old CD-1 ( ICR ) mice in standard cages or an EE comprising a running wheel , novel objects , and social interaction for 2 months . Motor and cognitive performances were evaluated using the rotarod test and passive avoidance test , and gene expression profile was investigated in the cerebral hemispheres using microarray and gene set enrichment analysis ( GSEA ) . RESULTS : In behavioral assessment , an EE significantly enhanced rotarod performance and short-term working memory . Microarray analysis revealed that genes associated with neuronal activity were significantly altered by an EE . GSEA showed that genes involved in synaptic transmission and postsynaptic signal transduction were globally upregulated , whereas those associated with reuptake by presynaptic neurotransmitter transporters were downregulated . In particular , both microarray and GSEA demonstrated that EE exposure increased opioid signaling , acetylcholine release cycle , and postsynaptic neurotransmitter receptors but decreased Na+ / Cl- -dependent neurotransmitter transporters , including dopamine transporter Slc6a3 in the brain . Western blotting confirmed that Q01959 , Q9UD71 ( Q9UD71 ) , and Q9H244 were largely altered in a region-specific manner . CONCLUSION : An EE enhanced motor and cognitive function through the alteration of synaptic activity-regulating genes , improving the efficient use of neurotransmitters and synaptic plasticity by the upregulation of genes associated with postsynaptic receptor activity and downregulation of presynaptic reuptake by neurotransmitter transporters . Clinical overview of promising nonthienopyridine antiplatelet agents . Three novel nonthienopyridine antiplatelet agents -- cangrelor , ticagrelor ( AZD6140 ) , and P35240 530348 -- are in advanced clinical testing in patients with coronary artery disease . DB06441 and ticagrelor are direct and reversible inhibitors of the platelet adenosine 5'-diphosphate Q9H244 receptor , whereas P35240 530348 is a thrombin receptor antagonist . Clinical data available to date for each of these compounds suggest that they have safety and efficacy profiles that will be advantageous to patients with acute coronary syndromes or undergoing percutaneous intervention . We review the clinical features of these new platelet inhibition therapies . Efficacy and safety of cangrelor for patients with coronary artery disease : a meta-analysis of four randomized trials . BACKGROUND : The efficacy and safety of new intravenous Q9H244 inhibitor ( cangrelor ) for patients with coronary artery disease ( CAD ) remain unclear . METHODS AND RESULTS : Trials were identified in PubMed , Web of Science , Embase , and Cochrane Database searches . We included four randomized , placebo-controlled reports in the meta-analysis . The database consisted of 36 , 081 patients on cangrelor compared with clopidogrel or placebo . Major adverse cardiac events ( MACE ) were defined as the primary efficacy endpoint and major or severe bleeding at 48 hours was defined as the primary safety endpoint . DB06441 significantly decreased risk of MACE ( OR : 0.87 , P = 0.002 ) and stent thrombosis ( OR : 0.53 , P < 0.001 ) . However , at the same time , an increase in TIMI minor bleeding ( OR : 1.49 , P = 0.04 ) and in GUSTO moderate bleeding ( OR : 1.43 , P = 0.04 ) were observed by cangrelor . CONCLUSIONS : Intravenous administration of cangrelor is benefit to reduce risk of MACE and stent thrombosis in patients with CAD excepting for increased minor bleeding events . The ' allosteric modulator ' P35240 -202676 disrupts G protein-coupled receptor function via sulphydryl-sensitive mechanisms . 1. Previous studies suggest that the thiadiazole compound P35240 -202676 ( N-(2,3-diphenyl-1,2,4-thiadiazol-5-(2H)-ylidene)methanamine ) acts as an allosteric modulator of a variety of structurally distinct G protein-coupled receptors ( GPCRs ) . It was postulated that P35240 -202676 would directly bind a structural motif in the receptor molecule common to divergent members of the GPCR family . The molecular mechanisms of such a promiscuous action , however , remain obscure . 2 . To clarify the mechanism of P35240 -202676 action , we used the functional approach of [35S]GTPgammaS autoradiography with rat brain cryostat sections together with classical membrane [35S]GTPgammaS binding assays to evaluate how the thiadiazole affects G protein activity mediated by various receptors linked to the Gi-family of G proteins . 3 . We found that in the absence of dithiotreitol ( DTT ) , P35240 -202676 ( 10(-7)-10(-5) M ) elicits nonspecific effects in the [35S]GTPgammaS-based G protein activation assays , thereby severely compromising interpretations on the compounds ability to allosterically inhibit receptor-mediated G protein activity . Such a nonspecific behaviour was fully reversed upon addition of DTT ( 1 mM ) , revealing thiol-based mechanism of action . 4 . In routine incubations containing DTT , P35240 -202676 had no effect on receptor-driven G protein activity , as assessed for adenosine A1 , alpha2-adrenergic , cannabinoid P21554 , lysophosphatidic acid Q92633 , muscarinic M2/M4 , purinergic Q9H244 or sphingosine 1-phosphate receptors , suggesting that the thiadiazole does not act as an allosteric modulator of GPCR function . 5 . 1H NMR analysis indicated that P35240 -202676 underwent structural changes after incubation with the reducing agent DTT or with brain tissue . 6 . We conclude that P35240 -202676 modulates GPCRs via thiol modification rather than via true allosteric mechanisms . Contribution of the Q9H244 receptor-mediated pathway to platelet hyperreactivity in hypercholesterolemia . BACKGROUND : In hypercholesterolemia , platelets demonstrate increased reactivity and promote the development of cardiovascular disease . OBJECTIVE : This study was carried out to investigate the contribution of the ADP receptor Q9H244 -mediated pathway to platelet hyperreactivity due to hypercholesterolemia . METHODS : P01130 -deficient mice and C57Bl/6 wild-type mice were fed on normal chow and high-fat ( Western or Paigen ) diets for 8 weeks to generate differently elevated cholesterol levels . Q9H244 receptor-induced functional responses via G(i) signaling were studied ex vivo when washed murine platelets were activated by 2MeSADP and PAR4 agonist AYPGKF in the presence and absence of indomethacin . Platelet aggregation and secretion , α(IIb)β(3) receptor activation and the phosphorylation of extracellular signal-regulated protein kinase ( P29323 ) and Akt were analyzed . RESULTS : Plasma cholesterol levels ranged from 69 ± 10 to 1011 ± 185 mg dL(-1) depending on diet in mice with different genotypes . Agonist-dependent aggregation , dense and α-granule secretion and JON/A binding were gradually and significantly ( P < 0.05 ) augmented at low agonist concentration in correlation with the increasing plasma cholesterol levels , even if elevated thromboxane generation was blocked . These functional responses were induced via increased levels of G(i) -mediated P29323 and Akt phosphorylation in hypercholesterolemic mice vs. normocholesterolemic animals . In addition , blocking of the Q9H244 receptor by AR-C69931MX ( DB06441 ) resulted in strongly reduced platelet aggregation in mice with elevated cholesterol levels compared with normocholesterolemic controls . CONCLUSIONS : These data revealed that the Q9H244 receptor pathway was substantially involved in platelet hyperreactivity associated with mild and severe hypercholesterolemia . DB06441 AstraZeneca . AstraZeneca is developing the P2T ( P2YADP ) purinoceptor antagonist and platelet aggregation inhibitor , cangrelor , for the potential treatment of unstable angina and as an ultrafast-acting intravenous antithrombotic agent . It is in phase IIb clinical trials [ 315723 ] . NDA and MAA applications are planned for after 2003 [ 275466 ] , [ 314472 ] . It superseded the earlier compound , ARL-67085 , which also reached phase II trials [ 328760 ] . In ex vivo samples of angina patients ' blood , cangrelor inhibits platelet/monocyte conjugate development , which indicate the drug has some degree of disease-modifying activity [ 377418 ] . AstraZeneca is also developing derivatives of cangrelor . Removal of the triphosphate side chain , modification of the ribose to a carbocycle and the purine to a triazolopyridine resulted in a potent ( IC50 = 4 nM ) orally-active P2T/ Q9H244 receptor antagonist . A lead compound was scheduled to enter trials as an antithrombotic agent in July 2000 [ 377666 ] . In March 1999 , Lehman Brothers predicted a 30 % probability that the drug would reach world markets and would be launched in 2002 [ 336599 ] . Electrocardiographic safety of cangrelor , a new intravenous antiplatelet agent : a randomized , double-blind , placebo- and moxifloxacin-controlled thorough QT study . DB06441 is an intravenous Q9H244 inhibitor under investigation as an antiplatelet drug in the setting of acute coronary syndromes . To determine the electrophysiologic safety of parenteral cangrelor , cardiac repolarization effects were measured in 67 healthy volunteers ( aged 18-45 years ) in a randomized crossover design , including 4 treatment sequences of therapeutic cangrelor , supratherapeutic cangrelor , placebo , and moxifloxacin ( positive control ) . Triplicate electrocardiogram measurements and pharmacokinetic samples were collected at baseline and 9 time points postdose on day 1 . For both cangrelor and moxifloxacin , time-matched , placebo-adjusted change in QT from baseline was evaluated using an individual ( QTcI ) heart rate correction . After cangrelor dosing , change in QTcI was < 5 ms at all times points and all corresponding upper 2-sided 90 % confidence intervals ( CIs ) were < 10 ms . Although moxifloxacin failed to show a lower CI > 5 ms , expected time trends and lower CI > 4.0 ms demonstrate assay sensitivity . QTcI was not affected by plasma concentrations of cangrelor metabolites , and cangrelor had no other adverse effects on electrocardiographic parameters . Clinically , cangrelor exposure was well tolerated . Thus , this thorough QT study demonstrated that therapeutic and supratherapeutic cangrelor doses do not adversely affect cardiac repolarization in normal volunteers ( clinicaltrials.gov ; identifier NCT00699504 ) . DB00820 , a further innovation in the treatment of sexual dysfunction . In recognition of the large number of sufferers of sexual dysfunction worldwide , and the variety of etiologies of the condition , investigation into effective pharmacological agents has been expanded . One method of intervention is inhibition of the phosphodiesterase type 5 ( O76074 ) enzyme , which has already been exploited with a considerable degree -- though not complete -- success . A number of new agents that inhibit O76074 are under development . Notable among these is tadalafil , which has demonstrated a high level of selectivity for O76074 over the other phosphodiesterases and has shown efficacy in improving erectile function and sexual satisfaction in phase III trials . Throughout the clinical development program for tadalafil , the drug has been well tolerated and without serious side effects . The manufacturer , Lilly Q9Y6W8 , received an approvable letter from the US Food and Drug Administration for use of the drug as a treatment for erectile dysfunction on April 30 , 2002 . Lilly Q9Y6W8 hopes to market tadalafil , with the trade name DB00820 , in the USA in 2003 . DB06441 for treatment of arterial thrombosis . INTRODUCTION : Percutaneous coronary intervention ( P05154 ) is a highly effective treatment for obstructive coronary artery disease . Oral platelet Q9H244 receptor antagonists reduce ischemic events in patients treated with P05154 . However , there are several limitations to their use , including variable pharmacodynamics , a slow onset and offset , and in those patients who are pretreated but subsequently require cardiac surgery , increased bleeding . DB06441 is an intravenous agent that provides rapid and intensive inhibition of the Q9H244 receptor that quickly dissipates after discontinuation . A recent , Phase III randomized clinical trial of P05154 patients demonstrated that cangrelor bolus and infusion reduced ischemic events compared with conventional clopidogrel therapy without increasing major bleeding . AREAS COVERED : This review outlines the pharmacodynamics , pharmacokinetics , and the safety and efficacy of cangrelor for the acute treatment of patients undergoing planned P05154 . EXPERT OPINION : DB06441 is an important addition to the current armamentarium of platelet inhibitors as it significantly reduces periprocedural myocardial infarction and stent thrombosis in a broad spectrum of patients , without increasing major bleeding or the need for transfusion . DB06441 will have particular benefit in clopidogrel-naïve patients with high anatomical complexity and/or increased clinical risk ( where the absolute risk for thrombotic and ischemic complications of P05154 is greatest ) . Pharmacodynamic effects during the transition between cangrelor and ticagrelor . OBJECTIVES : This study sought to determine pharmacodynamic effects during transition from intravenous cangrelor to oral ticagrelor and from oral ticagrelor to intravenous cangrelor . BACKGROUND : DB06441 is an intravenous antagonist of Q9H244 and its use will require transition to and from oral agents . METHODS : Patients ( n = 12 ) with stable coronary artery disease who were taking aspirin 81 mg daily were recruited . On study day 1 , they received a bolus plus 2-h infusion of cangrelor plus a 180-mg dose of ticagrelor at either 0.5 h ( n = 6 ) or 1.25 h ( n = 6 ) . Pharmacodynamic effects ( light transmission platelet aggregation in response to 20 and 5 μmol/l adenosine diphosphate , VerifyNow , Q9H244 assay ( Accumetrics , San Diego , California ) , vasodilator-stimulated phosphoprotein index , and flow cytometry ) were assessed during and after the cangrelor infusion . Patients took 90 mg of ticagrelor twice daily for either 6 ( n = 6 ) or 7 ( n = 6 ) doses . On study day 5 , pharmacodynamic effects were assessed before and during a bolus plus 2-h infusion of cangrelor . RESULTS : During cangrelor infusion , extensive inhibition of platelet function reflected by limited residual platelet reactivity was apparent . After cangrelor was stopped , the antiplatelet effects of ticagrelor were preserved despite a modest increase in platelet reactivity . CONCLUSIONS : DB08816 given before or during infusion of cangrelor did not attenuate the pharmacodynamic effects of cangrelor . The pharmacodynamic effects of ticagrelor were preserved when ticagrelor was given during infusion of cangrelor . Consistent with the reversible binding of ticagrelor , this oral Q9H244 antagonist can be administered before , during , or after treatment with cangrelor . DB06441 for patients undergoing percutaneous coronary intervention : evidence from a meta-analysis of randomized trials . DB06441 is a new parenteral adenosine diphosphate Q9H244 receptor inhibitor with rapid , profound and reversible inhibition of platelet activity . The aim of this meta-analysis was to evaluate efficacy and safety of this new agent in patients undergoing percutaneous coronary intervention ( P05154 ) . We searched PubMed , Cochrane Library , EMBASE , Web of Science and CINAHL databases from the inception through April 2013 . Randomized controlled trials ( RCTs ) comparing cangrelor with control ( clopidogrel/placebo ) were selected . We used the random-effects models to calculate the risk ratio . The primary efficacy outcome was risk of myocardial infarction , and the primary safety outcome was TIMI major bleeding at 48 h . Three RCTs included a total of 25,107 participants . Effects of DB06441 were not different against comparators for myocardial infarction ( MI ) ( Risk ratio [ RR ] 0.94 , 95 % confidence interval [ CI ] 0.78-1.13 ) and all-cause mortality ( RR 0.72 , 95 % CI 0.36-1.43 ) . However , cangrelor significantly reduced the risk of ischemia-driven revascularization ( RR 0.72 , 95 % CI 0.52-0.98 ) , stent thrombosis ( RR 0.60 , 95 % CI 0.44-0.82 ) and Q wave MI ( RR 0.53 , 95 % CI 0.30-0.92 ) without causing extra major bleeding ( Thrombolysis in Myocardial infarction criteria ) and severe or life-threatening bleeding ( Global utilization of streptokinase and tissue plasminogen activator for occluded coronary arteries criteria ) . Separate analysis against only clopidogrel also showed similar findings except Q wave MI outcome . Use of cangrelor during P05154 might reduce the risk of ischemia-driven revascularization and stent thrombosis , without causing extra major bleeding . DB06441 : review of the drug and the CHAMPION programme ( including PHOENIX ) . Platelet inhibition is the main goal of ancillary pharmacologic therapy during percutaneous coronary interventions ( P05154 ) . Thienopyridines and ticagrelor are oral drugs developed for this purpose . DB06441 is an intravenous , non-thienopyridine antagonist of the Q9H244 receptor with a rapid , potent , predictable , and quickly reversible effect . DB06441 has been studied in a broad population intended to receive P05154 in the CHAMPION program , where it was compared with different clopidogrel regimens . The first two trials , CHAMPION P05154 and PLATFORM , failed their primary objective , likely for challenges in the adjudication of P05154 -related myocardial infarction . In a third trial that implemented the universal definition of MI , CHAMPION PHOENIX , a reduction of thrombotic events , including stent thrombosis , was observed . In the BRIDGE trial cangrelor has been studied in patients who had to prematurely interrupt antiplatelet therapy for surgery . DB06441 appears a promising agent in patients who require P05154 or when a rapid reversal is needed . Differential role for phospholipase D1 and phospholipase D2 in 12-O-tetradecanoyl-13-phorbol acetate-stimulated MAPK activation , Cox-2 and P10145 expression . Phospholipase D ( PLD ) is expressed in many tissues and stimulated by growth factors and cytokines . However , the role of PLD in signal transduction is still not well-understood . Human embryonic kidney ( P29320 -293 ) cells exhibit low levels of both Q13393 and O14939 mRNA , however , only Q13393 protein was detected by Western blot . When either isoform of PLD was stably expressed in P29320 -293 cells , we observed an increased PLD activity in a cell-free system and a 12-O-tetradecanoyl-13-phorbol acetate ( TPA ) -stimulated increase in PLD activity in intact cells . This system was then used to elucidate the effects of PLD activity on TPA-stimulated signaling pathways . Two such pathways , the mitogen-activated protein kinases ( MAPK ) , extracellular regulated protein kinase ( P29323 ) and p38 are activated by growth factors and cellular stress , respectively . We found that TPA stimulated P29323 phosphorylation regardless of the expression status of PLD . In contrast to P29323 kinase , P29320 -293 cells were unable to induce p38 phosphorylation by TPA stimulation . When P29320 -293 cells expressed either Q13393 or O14939 , we observed elevated p38 phosphorylation in response to TPA stimulation . The P29323 and p38 MAPKs can also stimulate the expression of both cyclooxygenase-2 ( Cox-2 ) and interleukin-8 ( P10145 ) . We used this system to differentiate the effect of Q13393 or O14939 activity on the expression of Cox-2 and P10145 . Increased Cox-2 and P10145 expression was found only in P29320 -293 cells expressing Q13393 . These data identify a novel role for the Q13393 isoform in the induction of gene expression and provide new insight into the differential role of Q13393 and O14939 in cells . Potentiation of platelet aggregation by heparin in human whole blood is attenuated by Q9H244 and P47900 antagonists but not aspirin . INTRODUCTION : DB01109 ( UFH ) potentiates platelet aggregation induced by some agonists . Q9H244 and P47900 receptors play a major role in amplifying platelet aggregation . We assessed the ability of cangrelor , a selective Q9H244 antagonist , A2P5P , a selective P47900 antagonist , and aspirin to block the potentiating effects of heparin . MATERIALS AND METHODS : Whole blood from healthy human volunteers was anticoagulated with either hirudin or UFH 10 IU/ml . Some tubes anticoagulated with hirudin also contained UFH 1 or 10 IU/ml . The low-molecular-weight heparin dalteparin was also assessed . Platelet aggregation was performed using whole blood single-platelet counting . Dense granule release was assessed using 14C-5HT-labelled platelets . RESULTS : UFH and , to a lesser extent , dalteparin potentiated platelet aggregation induced by ADP , Q15004 , 5HT , U46619 , epinephrine and TRAP in a concentration-dependent manner but inhibited aggregation induced by collagen . DB06441 effectively opposed the potentiating effects of heparins on sustained aggregation induced by ADP , Q15004 , 5HT , U46619 and TRAP but had less effect on epinephrine-induced aggregation , whereas A2P5P was more effective at blocking both the initial phase of ADP-induced aggregation and the aggregation response to epinephrine , reflecting the differences in G protein coupling between the agonist receptors . DB00945 had no effect on potentiation by heparin . Heparins did not increase ADP- or TRAP-induced 14C-5HT release . CONCLUSIONS : Heparins potentiate platelet responses to ADP and numerous other agonists . This potentiation is attenuated by cangrelor and A2P5P , and is not mediated by increased dense granule release . ADP receptor antagonists but not aspirin may have potential therapeutic benefits in counteracting the pro-thrombotic effects of heparins . Emerging Q9H244 receptor antagonists : role in coronary artery disease . The use of oral antiplatelet therapy in reducing vascular events has been extensively studied . Currently available oral antiplatelet agents include aspirin and the thienopyridine Q9H244 receptor antagonists . These classes are combined frequently in the setting of acute coronary syndrome and percutaneous coronary intervention ( P05154 ) . Resistance to either or both of these agents is a major concern , as antiplatelet resistance has been linked to an increase in thrombotic events and worse clinical outcomes . As a result , there is a need for newer , more effective antiplatelet agents to address the limitations of currently available therapy . Prasugrel , a third generation thienopyridine , has been approved by both the FDA and European Commission . Two additional Q9H244 agents , ticagrelor and cangrelor are in advanced stages of development . The possible advantages of prasugrel over clopidogrel include a faster onset of action , reduced inter-patient variability and more potent platelet inhibition . DB08816 is an oral reversible Q9H244 antagonist with greater platelet inhibition compared with clopidogrel . DB06441 is being developed as an intravenous Q9H244 antagonist with a very fast onset and offset , which may offer advantages particularly in the setting of coronary intervention . These emerging antiplatelet agents may offer advantages such as faster onset of action , greater potency and reversibility of platelet inhibition . This article summarizes the available clinical data on the upcoming Q9H244 antiplatelet agents in the treatment of coronary artery disease .

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Pharmacological properties of thalidomide and its analogues . Thalidomide and its immunomodulatory imide drugs ( IMiDs ) analogues DB00480 ( DB00480 , DB00480 ) and CC-4047 ( Actimid , DB08910 ) have been used as anti-inflammatory and anticancerous drugs in the recent years . Thalidomide and IMiDs inhibit the cytokines tumour necrosis factor-alpha ( P01375 ) , interleukins ( IL ) 1-beta , 6 , 12 , and granulocyte macrophage-colony stimulating factor ( GM- P04141 ) . They also costimulate primary human T , NKT and NK lymphocytes inducing their proliferation , cytokine production , and cytotoxic activity . On the other hand , the compounds are anti-angiogenic , anti-proliferative , and pro-apoptotic . Thalidomide analogues have been used as inhibitors of alpha glucosidase and could be potential drugs for diabetes treatment . In this review , we explore the current trend of the different structures , the new patents , and the possible new applications in different pathologies . Cellular distribution and contribution of cyclooxygenase P35354 to diabetogenesis in NOD mouse . Unlike most other mammalian cells , beta-cells of Langerhans constitutively express cyclooxygenase ( P36551 ) -2 rather than P23219 . P35354 is also constitutively expressed in type 1 diabetes ( T1D ) patients ' periphery blood monocytes and macrophage . To understand the role of P35354 in the beta-cell , we investigated P35354 expression in beta-cells and islet infiltrates of NOD and BALB/c mice using fluorescence immunohistochemistry and cytochemical confocal microscopy and Western blotting . Immunostaining showed that P35354 is expressed in islet-infiltrating macrophages , and that the expression of insulin and P35354 disappeared concomitantly from the beta-cells when NOD mice progressed toward overt diabetes . Also cultured P01308 -1E cells coexpressed insulin and P35354 but clearly in different subcellular compartments . Treatment with celecoxib increased insulin release from these cells in a dose-dependent manner in glucose concentrations ranging from 5 to 17 mM . Excessive P35354 expression by the islet-infiltrating macrophages may contribute to the beta-cell death during insulitis . The effects of celecoxib on P01308 -1E cells suggest that PGE(2) and other downstream products of P35354 may contribute to the regulation of insulin release from the beta-cells . Ca2+-calmodulin and janus kinase 2 are required for activation of sodium-proton exchange by the Gi-coupled 5-hydroxytryptamine 1a receptor . The type 1 sodium-proton exchanger ( P19634 ) is expressed ubiquitously and regulates key cellular functions , including mitogenesis , cell volume , and intracellular pH . Despite its importance , the signaling pathways that regulate P19634 remain incompletely defined . In this work , we present evidence that stimulation of the 5-hydroxytryptamine 1A ( P08908 ) receptor results in the formation of a signaling complex that includes activated O60674 ( Jak2 ) , Ca2+/calmodulin ( P62158 ) , and P19634 , and which involves tyrosine phosphorylation of P62158 . The signaling pathway also involves rapid agonist-induced association of P62158 and P19634 as assessed by coimmunoprecipitation studies and by bioluminescence resonance energy transfer studies in living cells . We propose that P19634 is activated through this pathway : P08908 receptor --> G(i2)alpha and/or G(i3)alpha --> Jak2 activation --> tyrosine phosphorylation of P62158 --> increased binding of P62158 to P19634 --> induction of a conformational change in P19634 that unmasks an obscured proton-sensing and/or proton-transporting region of P19634 --> activation of P19634 . The G(i/o)-coupled P08908 receptor now joins a handful of Gq-coupled receptors and hypertonic shock as upstream activators of this emerging pathway . In the course of this work , we have presented clear evidence that P62158 can be activated through tyrosine phosphorylation in the absence of a significant role for elevated intracellular Ca2+ . We have also shown for the first time that the association of P62158 with P19634 in living cells is a dynamic process . Lipopolysaccharide directly stimulates cortisol secretion by human adrenal cells by a cyclooxygenase-dependent mechanism . Activation of the hypothalamo-pituitary-adrenal axis by bacterial lipopolysaccharide ( LPS ; endotoxin ) is well documented , although there has been uncertainty about whether LPS exerts a direct effect at the level of the adrenal . The present study found that LPS caused a dose-dependent stimulation of basal cortisol secretion by the human adrenocortical cell line , NCI-H295R , without affecting aldosterone . The expression of both O60603 ( O60603 ) and O00206 was demonstrated in these cells , and the specific ligands for O00206 ( purified LPS and lipid A ) and O60603 ( Pam3Cys ) were found to stimulate cortisol release , suggesting that these receptors may mediate the effects of LPS in adrenal cells , as has been shown in other cell types . LPS was also found to stimulate prostaglandin E2 release by these cells . The effects of LPS on cortisol were attenuated in the presence of both indomethacin and a specific P35354 inhibitor , but not a P23219 inhibitor , suggesting an obligatory role for P35354 activation and prostaglandin synthesis in the adrenal response to LPS . Signaling pathways mediating induction of the early response genes prostaglandin G/H synthase-2 and egr-1 by serotonin via 5- Q13049 receptors . Signaling pathways responsible for serotonin ( 5-HT ) -mediated induction of early response genes prostaglandin G/H synthase-2 ( P35354 , cyclooxygenase-2 ) and egr-1 were investigated in rat mesangial cells . Gene induction by 5-HT was dependent on 5- Q13049 receptors that were pertussis toxin insensitive indicating coupling to a G-protein of the Gq family . Binding of 5-HT to this receptor activates phosphatidylinositol-specific phospholipase C ( P98160 ) and release of Ca2+ from internal stores , but this activation was not related to P35354 mRNA expression . Similarly , P19957 kinase was not involved in 5-HT signaling . Instead , inhibition of phosphatidylcholine-specific P98160 interfered with P35354 and egr-1 mRNA induction , suggesting this enzyme as a link between 5- Q13049 receptors and protein kinase C , an essential part of 5-HT-mediated signaling . The Q96HU1 kinase pathway was identified as common signaling pathway of 5-HT or phorbol ester-induced gene expression . Increase of intracellular DB02527 by forskolin or dibutyryl DB02527 did not induce P35354 or egr-1 mRNA expression by itself , but strongly inhibited 5-HT-mediated mRNA induction . P35354 mRNA and protein induction by 5-HT was also abolished by chelation of Ca2+ ions by EGTA , suggesting involvement of Ca2+-dependent enzymes . In contrast , egr-1 mRNA expression was superinduced in the presence of EGTA . Induction of Egr-1 protein was not changed by EGTA hinting to Ca2+-sensitive posttranscriptional steps . Activation of the Gq-coupled 5- Q13049 receptor thus leads to the expression of the early response genes P35354 and egr-1 , using common as well as differing signaling elements that allow differential regulation of the expression of these genes that are functionally related to renal hemodynamics and proliferation of mesangial cells , respectively . The role of endothelium-derived hyperpolarizing factor and cyclooxygenase pathways in the inhibitory serotonergic response to the pressor effect elicited by sympathetic stimulation in chronic sarpogrelate treated rats . We have demonstrated that the antagonism of 5-HT2 receptors produces an enhancement of serotonergic sympathoinhibitory effect by P28221 and P34969 activation . The aim of this work was to determine mechanisms involved in the 5-hydroxytriptaminergic inhibitory action on the pressor responses elicited by sympathostimulation in pithed rats treated with a 5-HT2 receptor blocker . The blockade of 5-HT2 receptors was induced by orally sarpogrelate treatment ( 30 mg/kg/day ) . Two weeks later , animals were anaesthetized and pithed . A bolus injection of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one ( ODQ ) ( 10 µg/kg ) , a guanylyl cyclase inhibitor , or indomethacin ( 2mg/kg ) , a non-selective P36551 inhibitor , prior to the infusion of ( 2S ) (+)-5-(1,3,5-trimethylpyrazol-4-yl)-2-(dimethylamino)tetralin , AS-19 ( 5 µg/kg/min ) were not able to abolish its inhibitory action . However , i.v. administration of glibenclamide ( 20mg/kg ) , a blocker of DB00171 -sensitive K(+) channels , completely reversed AS-19 sympathoinhibitory action . The inhibitory effect of 2-[5-[3-(4-methylsulfonylamino)benzyl-1,2,4-oxadiazol-5-yl]-1H-indol-3-yl]ethanamine , L-694,247 ( 5 µg/kg/min ) was abolished by indomethacin , whereas pretreatment with ODQ had no effect . DB04743 ( 3mg/kg ) , a P35354 inhibitor , completely reversed the inhibitory action of L-694,247 , whereas 1- [ [ 4,5-bis (4-methoxyphenyl)-2-thiazolyl ] carbonyl ] -4-methylpiperazine hydrochloride ( FR122047 ) ( 3mg/kg ) , a P23219 inhibitor , partially blocked this action . The sympathoinhibition by 5-HT ( 20 µg/kg/min ) could not be elicited after i.v. treatment with indomethacin plus glibenclamide . In conclusion , these results suggest that in chronic sarpogrelate-treated rats , the inhibitory serotonergic effect of the pressor responses induced by electrical stimulation of the sympathetic outflow via P34969 and P28221 receptor activation is mediated by KATP channel-mediated smooth muscle hyperpolarization and the P36551 pathway , respectively . Substance P stimulates cyclooxygenase-2 and prostaglandin E2 expression through JAK- P35610 activation in human colonic epithelial cells . Substance P ( SP ) via its neurokinin-1 receptor ( P25103 ) regulates several gastrointestinal functions . We previously reported that P25103 -mediated chloride secretion in the colon involves formation of PG . DB00917 biosynthesis is controlled by cyclooxygenase-1 ( P23219 ) and P35354 , whose induction involves the STATs . In this study , we examined whether SP stimulates DB00917 production and P35354 expression in human nontransformed NCM460 colonocytes stably transfected with the human P25103 ( NCM460- P25103 cells ) and identified the pathways involved in this response . SP exposure time and dose dependently induced an early ( 1-min ) phosphorylation of O60674 , P40763 , and P42229 , followed by P35354 expression and DB00917 production by 2 h . Pharmacologic experiments showed that DB00917 production is dependent on newly synthesized P35354 , but P23219 protein . Inhibition of protein kinase Ctheta ( PKCtheta ) , but not PKCepsilon and PKCdelta , significantly reduced SP-induced P35354 up-regulation , and O60674 , P40763 , and P42229 phosphorylation . Pharmacological blockade of JAK inhibited SP-induced O60674 , P40763 , and P42229 phosphorylation ; P35354 expression ; and DB00917 production . Transient transfection with O60674 short-interferring RNA reduced P35354 promoter activity and O60674 phosphorylation , while RNA interference of P35610 isoforms showed that P42229 predominantly mediates SP-induced P35354 promoter activity . Site-directed mutation of P35610 binding sites on the P35354 promoter completely abolished P35354 promoter activity . Lastly , P35354 expression was elevated in colon of mice during experimental colitis , and this effect was normalized by administration of the P25103 antagonist CJ-12,255 . Our results demonstrate that SP stimulates P35354 expression and DB00917 production in human colonocytes via activation of the O60674 - P40763 /5 pathway . n-3 fatty acids specifically modulate catabolic factors involved in articular cartilage degradation . This study describes specific molecular mechanisms by which supplementation with n-3 fatty acids ( i.e. those present in fish oils ) can modulate the expression and activity of degradative and inflammatory factors that cause cartilage destruction during arthritis . Our data show that incorporation of n-3 fatty acids ( but not other polyunsaturated or saturated fatty acids ) into articular cartilage chondrocyte membranes results in a dose-dependent reduction in : ( i ) the expression and activity of proteoglycan degrading enzymes ( aggrecanases ) and ( ii ) the expression of inflammation-inducible cytokines ( interleukin ( IL ) -1alpha and tumor necrosis factor ( P01375 ) -alpha ) and cyclooxygenase ( P35354 ) , but not the constitutively expressed cyclooxygenase P23219 . These findings provide evidence that n-3 fatty acid supplementation can specifically affect regulatory mechanisms involved in chondrocyte gene transcription and thus further advocate a beneficial role for dietary fish oil supplementation in alleviation of several of the physiological parameters that cause and propogate arthritic disease . Toll-like receptor expression in human keratinocytes : nuclear factor kappaB controlled gene activation by Staphylococcus aureus is toll-like receptor 2 but not toll-like receptor 4 or platelet activating factor receptor dependent . Cultured primary human keratinocytes were screened for their expression of various members of the toll-like receptor ( TLR ) family . Keratinocytes were found to constitutively express Q15399 , O60603 , O15455 , O60602 , and Q9NR96 but not O00206 , Q9Y2C9 , Q9NYK1 , Q9NR97 , or Q9BXR5 as shown by polymerase chain reaction analysis . The expression of the crucial receptor for signaling of staphylococcal compounds O60603 was also confirmed by immunohistochemistry , in contrast to O00206 , which showed a negative staining pattern . Next , we analyzed the activation of the proinflammatory nuclear transcription factor kappaB by Staphylococcus aureus strain 8325-4 . Using nuclear extract gel shifts , RelA staining , and luciferase reporter transfection plasmids we found a clear induction of nuclear factor kappaB translocation by the bacteria . This translocation induced the transcription of nuclear factor kappaB controlled genes such as inducible nitric oxide synthetase , P35354 , and interleukin-8 . Transcription of these genes was followed by production of increased amounts of interleukin-8 protein and NO . Inhibition experiments using monoclonal antibodies and the specific platelet activating factor receptor inhibitor CV3988 showed that nuclear factor kappaB activation by S. aureus was O60603 but not O00206 or platelet activating factor receptor dependent . In line , the purified staphylococcal cell wall components lipoteichoic acid and peptidoglycan , known to signal through O60603 , also showed nuclear factor kappaB translocation in human keratinocytes , indicating a crucial role of the staphylococcal cell wall in the innate immune stimulation of human keratinocytes . These results help to explain the complex activation of human keratinocytes by S. aureus and its cell wall components in various inflammatory disorders of the skin . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Regulatory regions of growth-related genes can activate an exogenous gene of the alpha-fetoprotein promoter to a comparable degree in human hepatocellular carcinoma cells . We examined the transcriptional activation by the regulatory regions of the midkine ( MK ) , survivin ( Q09428 ) , cyclooxygenase-2 ( P35354 ) , telomerase reverse transcriptase ( O14746 ) and alpha-fetoprotein ( AFP ) genes in human hepatocellular carcinoma cells . Luciferase assays showed that the Q09428 regulatory region exhibited the greatest activity and that the MK regulatory region activated the reporter gene better than the enhancer-linked AFP promoter even in high-AFP-producing cells . The P35354 and O14746 regulatory regions also activated the reporter gene better than the AFP enhancer/promoter in intermediate-AFP-producing cells . Combination of the regulatory regions arranged in tandem modulated their transcriptional activities , depending on the arrangement of the promoters and cells examined . These data suggested that the regulatory regions of the growth-related genes could be useful to activate a therapeutic gene in hepatocellular carcinoma cells irrespective of the amounts of AFP production but combinatory use of the promoter regions could not always contribute to enhanced activity . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . Maximizing clinical benefit with trastuzumab . To optimize patient management in breast cancer a number of factors are considered , including hormone receptor and P04626 status . A feasible approach for women with less aggressive , estrogen receptor/ P04626 -positive metastatic breast cancer is to consider trastuzumab ( Herceptin ; F. Hoffmann-La Roche , Basel , Switzerland ) combined with endocrine therapy . Randomized clinical trials are ongoing to assess the combination of trastuzumab with aromatase inhibitors . In patients with aggressive P04626 -positive metastatic breast cancer , trastuzumab/chemotherapy combination regimens are warranted . When administered first line in combination with a taxane , trastuzumab improves all clinical outcome parameters , including survival , in such patients . DB00072 adds little to the toxicity profile of taxanes , and trastuzumab combination therapy is associated with improvements in quality of life when compared with chemotherapy alone . There is encouraging evidence of improved efficacy when trastuzumab is combined with other cytotoxic agents with proven single-agent activity in breast cancer , including capecitabine ( DB01101 ; F. Hoffmann-La Roche ) , gemcitabine , and vinorelbine . DB00072 is also being investigated as part of triplet drug regimens . DB00072 has good single-agent activity in first-line therapy . This is of relevance to women with P04626 -positive disease who are not suitable for , or do not wish to receive , cytotoxic chemotherapy . The benefits noted with trastuzumab-containing regimens were documented in clinical trials where trastuzumab was given until disease progression . A further rationale exists to continue trastuzumab beyond progression . Data from retrospective reviews indicate that this strategy is feasible . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Dietary conjugated linoleic acid enhances spleen P37231 mRNA expression in broiler chicks . 1. The anti-inflammatory effects of dietary conjugated linoleic acid ( DB01211 ) on broilers repeatedly challenged with lipopolysaccharide ( LPS ) were investigated . 2 . Day-old broiler chicks were allotted into three treatment groups and fed on a control diet or diets containing 5.0 or 10.0 g DB01211 /kg diet . Six chicks from each treatment were injected with LPS ( 0.25 mg/kg body weight ) at 16 , 18 and 20 d of age . Splenic cyclooxygenase ( P36551 ) and inducible nitric oxide synthase ( P35228 ) activities , and prostaglandin E(2) ( PGE(2) ) and nitric oxide ( NO ) production as well as peroxisome proliferator-activated receptor-gamma ( P37231 ) mRNA expression were measured at 21 d of age . 3 . Chicks fed 10.0 g DB01211 /kg diet had lower P36551 activities and PGE(2) production that the controls . Dietary DB01211 ( 10.0 g/kg ) did not significantly diminish LPS-induced enhancement of COS-2 activity , inhibited the subsequent increase in PGE(2) production . 4 . Regulation of P23219 activity contributed to the difference in PGE(2) production . 5 . DB01211 did not markedly attenuate the increase of P35228 activity and NO production caused by LPS challenge . Chicks fed DB01211 had lower P35228 activity and NO production than the controls . 6 . Dietary DB01211 activated splenic P37231 mRNA expression and increased P37231 mRNA expression after LPS injection . 7 . These results suggest that dietary DB01211 has immunomodulatory effects in the spleen by restricting basal PGE(2) and NO to lower levels and enhancing P37231 mRNA expression . During the inflammatory response , dietary DB01211 did not alleviate the increase in P35354 and P35228 activities but enhanced Q06203 -gamma mRNA expression . P00797 inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE(-/-) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE(-/-) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 also decreased the levels of AT1R , gp91phox , O60603 , monocyte chemotactic protein-1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE(-/-) mice , but aliskiren showed no further benefits in ApoE(-/-) CatS(-/-) mice . In vitro , O60603 silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or/and ex vivo . CONCLUSION : P00797 inhibition appears to inhibit advanced plaque neovessel formation in ApoE(-/-) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R/ O60603 -mediated CatS activation and activity , thus regressing advanced atherosclerosis . aChE and BuChE inhibition by rivastigmin have no effect on peripheral insulin resistance in elderly patients with Alzheimer disease . BACKGROUND : P01308 resistance ( IR ) may play a role in most pathogenic processes that promote the development of Late Onset Alzheimer Disease ( LOAD ) . This study was designed to determine the interaction between inhibition of both butyrylcholinesterase ( BuChE ) and acetylcholinesterase ( P22303 ) with rivastigmine and peripheral insulin resistance ( IR ) in LOAD . METHODS : Seventy-Nine consecutive elderly patients , 31 late onset AD and 48 non-demented patients were evaluated . IR was calculated with HOMA . All of the patients were evaluated through comprehensive geriatric assessments at baseline and in the 6th and 12th months . RESULTS : End of the study , compared to the baseline values , there was a significant increase in the 6th month in both MMSE and IADL scores ( t =2.200 , p = 0.036 for MMSE and t =2.724 , p= 0.011 for IADL , respectively ) . DB00989 was improved both the scores of MMSE and IADL in elderly patients with LOAD , but there was no significance or correlation between HOMA scores and cognitive status . CONCLUSION : In conclusion , inhibition of both BuChE and P22303 with rivastigmine was improved the cognition without affecting on the peripheral IR in the elderly patients with LOAD by HOMA . Due to the complexity of disease pathogenesis , it is too early to make general comments , and further longitudinal and long-term studies on this issue are needed . Augmentation by citalopram of risperidone-induced monoamine release in rat prefrontal cortex . RATIONALE : A typical antipsychotics ( APDs ) , e.g. olanzapine and risperidone , have been reported to be effective adjunctive treatment for depression if selective serotonin ( 5-HT ) reuptake inhibitors ( SSRIs ) alone are ineffective . OBJECTIVES AND METHODS : We utilized microdialysis in awake , freely moving rats to study the effect of risperidone in combination with citalopram , an SSRI , on extracellular 5-HT , dopamine ( DA ) , and norepinephrine ( NE ) efflux in rat medial prefrontal cortex ( mPFC ) . RESULTS : DB00734 ( 1.0 mg/kg , s.c. ) , given alone , significantly increased 5-HT , DA , and NE concentrations in the mPFC . DB00215 ( 10 mg/kg , s.c. ) , by itself , produced a significant increase in 5-HT levels only . The combination of risperidone and citalopram produced significantly greater increases in efflux of both DA and NE than risperidone alone . However , the effect of this combination on extracellular 5-HT concentrations was not significantly different than that of citalopram alone . The augmentation of DA and NE efflux induced by risperidone plus citalopram could be partially blocked by the selective P08908 antagonist , WAY 100635 ( 0.2 mg/kg , s.c. ) . CONCLUSIONS : The results suggest that the ability of atypical APDs to augment the therapeutic efficacy of SSRIs in major depression and treatment-resistant depression may be due , at least in part , to potentiation of SSRI-induced increases in cortical DA and NE . The contributions of P08908 receptor stimulation and 5- Q13049 and alpha2 adrenergic receptor antagonism to this augmentation are discussed . Antiinflammatory effect of lactic acid bacteria : inhibition of cyclooxygenase-2 by suppressing nuclear factor-kappaB in Raw264.7 macrophage cells . Lactobacillus casei 3260 ( L. casei 3260 ) was evaluated in relation to the inflammatory response mediated by lipopolysaccharide ( LPS ) -induced nuclear factor-kappaB ( NF-kappaB ) and cyclooxygenase-2 ( P35354 ) expression in Raw264.7 macrophage cells . The treatment of Raw264.7 cells with L. casei 3260 significantly inhibited the secretion of tumor necrosis factor-alpha ( P01375 ) and prostaglandins E2 ( DB00917 ) , followed by suppression of P35354 . To clarify the molecular mechanism , the inhibitory effect of L. casei 3260 on the NF-kappaB signaling pathway was examined based on the luciferase reporter activity . Although the treatment of Raw264.7 cells with L. casei 3260 did not affect the transcriptional activity of NF-kappaB , it did inhibit NF-kappaB activation , as determined by the cytosolic p65 release and degradation of I-kappaBalpha . Therefore , these findings suggest that the suppression of P35354 through inhibiting the NF-kappaB activation by LPS may be associated with the antiinflammatory effects of L. casei 3260 on Raw264.7 cells . Meloxicam . Meloxicam ( DB00814 trade mark , Boehringer Ingelheim ) is a relatively new oral non-steroidal anti-inflammatory drug ( NSAID ) approved for the treatment of osteoarthritis in the US . It has also been evaluated for the treatment of rheumatoid arthritis , ankylosing spondylitis and acute ' rheumatic ' pain . Meloxicam has been shown to be P35354 preferential , particularly at its lowest therapeutic dose , and is anti-inflammatory by inhibiting prostanoid synthesis in inflammatory cells . Since it is P35354 preferential , it would be expected to have less gastrointestinal toxicity than non-selective NSAIDs . In clinical trials of meloxicam in osteoarthritis , it was found to be as effective as piroxicam , diclofenac and naproxen with less clinical gastrointestinal symptoms and less perforations , obstructions and bleeds by meta-analysis . Adverse events , including peripheral oedema and hypertension , occurred at a similar rate as with traditional NSAIDs . Effects of P04626 amplicon size and genomic alterations of chromosomes 1 , 3 , and 10 on patient response to trastuzumab in metastatic breast cancer . DB00072 is widely used for advanced breast cancer patients with P04626 -amplified tumors . Nevertheless , over half of these patients do not have an objective response . One reason may be altered expression of genes that might compensate for P04626 inhibition . We previously mapped the gene-rich region of chromosome 17 telomeric to P04626 , and reported considerable variability in the telomeric extent of the P04626 amplicon . Here we examined whether the variable amplicon size may be associated with patient response to trastuzumab . In addition , we looked at associations between response and several signaling pathway-related genes unrelated to the P04626 amplicon , including Q9Y243 , P60484 , P42336 , and P35354 . In 35 patients with P04626 -amplified metastatic breast cancer , with 40 % overall response to trastuzumab , fluorescence in situ hybridization identified the telomeric extent of the P04626 amplicon and the status of the several pathway-related genes . Objective response strongly correlated with the telomeric amplicon size , with 62 % of patients with shorter amplicons responding , compared with only 7 % of patients with longer amplicons ( P = 0.0015 ) . Abnormal copy number of P35354 was marginally associated with objective response ( P = 0.066 ) , while abnormal copy numbers of two reference loci , 1q25 and the chromosome 10 centromere , were significantly associated with response . Pairwise combinations of copy number status of these loci and P04626 amplicon size provided stronger associations and identified a group of patients without responders . These results suggest that patient selection for trastuzumab may be improved by considering P04626 amplicon size and genomic status of the 1q25 , P35354 , and centromere 10 loci . DB08877 for the treatment of primary myelofibrosis . PURPOSE : The pharmacology , pharmacokinetics , pharmacogenomics , clinical efficacy , and safety profile of ruxolitinib for the treatment of primary myelofibrosis are reviewed . SUMMARY : DB08877 , an oral tyrosine kinase inhibitor that targets the Janus-associated kinases ( JAKs ) 1 and 2 , has been recently approved for the treatment of patients with intermediate- or high-risk myelofibrosis . Unlike previous treatment options for patients with myelofibrosis , ruxolitinib offers a targeted therapy option for these patients who often suffer with severe and debilitating symptoms associated with the disease process . After oral administration , ruxolitinib is rapidly absorbed and can be given without regard to meals . DB08877 is primarily metabolized by the cytochrome P-450 ( CYP ) 3A4 isoenzyme system ; therefore , if concomitant use with a strong P08684 inhibitor is unavoidable , an initial dosage reduction is warranted . Two Phase III randomized trials comparing ruxolitinib to either placebo or best available therapy found a rapid and sustained response in the reduction of spleen size and improvements in constitutional symptoms and quality of life , with one study demonstrating an improvement in overall survival . The most commonly reported serious adverse effects of ruxolitinib are anemia and thrombocytopenia . DB08877 is administered as an oral tablet given twice daily , with the initial starting dosage based on the baseline platelet count . Dosage reductions are based on the development of thrombocytopenia . CONCLUSION : By directly targeting both P23458 and O60674 through small-molecule inhibition , ruxolitinib elicits a reduction in splenomegaly and disease-related symptoms in patients with intermediate- or high-risk myelofibrosis while maintaining an acceptable toxicity profile and a low treatment-discontinuation rate . Bioassay-guided isolation of an alkaloid with antiangiogenic and antitumor activities from the extract of Fissistigma cavaleriei root . Fissistigma cavaleriei ( Levl ) Rehd ( Annonaceae ) is used as a folklore medicine for treatment of inflammation , arthritis , and tuberculosis by Miao people in China . In the present study , the antiangiogenic activity of F. cavaleriei was investigated . The chorioallantoic membrane of the fertilized hen 's egg ( P62158 assay ) was used to determine antiangiogenic activity of the plant extract . Compound ( 1 ) , a compound with antiangiogenic activity , was isolated by bioassay-guided fractionation from F. cavaleriei for the first time . The structure of compound ( 1 ) was elucidated on the basis of spectroscopic methods . Colorimetric P36551 ( ovine ) inhibitor screening assay was used to determine its inhibitory effect on P23219 and P35354 . MTT and Sulforhodamine B assays were used to investigate its cytotoxic effects on tumor cell lines . As a result , compound ( 1 ) showed a selectively inhibiting effect on P35354 and could inhibit the growth of tumor cells in vitro . The antitumor activity of compound ( 1 ) was further confirmed by the observation that compound ( 1 ) administration significantly inhibited the growth of S-180 cells in mice . Moreover , compound ( 1 ) was able to enhance the antitumor activity of doxorubicin in the mice bearing with S-180 cells while combined with doxorubicin . In conclusion , compound ( 1 ) is a multi-target molecule and further experimental investigations are needed to determine whether it can be used as a lead molecule for tumor treatment . Involvement of 5-HT₇ receptors in vortioxetine 's modulation of circadian rhythms and episodic memory in rodents . Since poor circadian synchrony and cognitive dysfunction have been linked to affective disorders , antidepressants that target key 5-HT ( serotonin ) receptor subtypes involved in circadian rhythm and cognitive regulation may have therapeutic utility . DB09068 is a multimodal antidepressant that inhibits P28221 , 5- Q9H205 , P34969 receptor activity , 5-HT reuptake , and enhances the activity of P08908 and P28222 receptors . In this study , we investigated the effects of vortioxetine on the period length of O15055 ::LUC expression , circadian behavior , and episodic memory , using tissue explants from genetically modified O15055 ::LUC mice , locomotor activity rhythm monitoring , and the object recognition test , respectively . Incubation of tissue explants from the suprachiasmatic nucleus of O15055 ::LUC mice with 0.1 μM vortioxetine increased the period length of O15055 bioluminescence . Monitoring of daily wheel-running activity of Sprague-Dawley rats treated with vortioxetine ( 10 mg/kg , s.c. ) , alone or in combination with the P08908 receptor agonist flesinoxan ( 2.5 mg/kg , s.c. ) or the P34969 receptor antagonist SB269970 ( 30 mg/kg , s.c. ) , just prior to activity onset revealed significant delays in wheel-running behavior . The increase in circadian period length and the phase delay produced by vortioxetine were abolished in the presence of the P34969 receptor partial agonist AS19 . Finally , in the object recognition test , vortioxetine ( 10 mg/kg , i.p. ) increased the time spent exploring the novel object during the retention test and this effect was prevented by AS19 ( 5 mg/kg , i.p. ) . In conclusion , the present study shows that vortioxetine , partly via its P34969 receptor antagonism , induced a significant effect on circadian rhythm and presented promnesic properties in rodents . Proteomic profiling of cancer stem cells derived from primary tumors of P04626 /Neu transgenic mice . Human epidermal growth factor receptor 2 ( P04626 ) overexpression leads to mammary tumorigenesis and its elevated levels lead to increase in cancer stem cells ( CSCs ) , invasion , and metastasis . CSCs are resistant to radiation/chemotherapeutic drugs and are believed to be responsible for recurrence/relapse of cancer . CSCs are isolated using flow cytometry based sorting , although reliable , this technology hinders the convenient identification of molecular targets of CSCs . Therefore to understand the molecular players of increased CSC through P04626 overexpression and to develop meaningful targets for combination therapy , we isolated and characterized breast CSCs through convenient tumorsphere culture . We identified the altered protein expression in CSC as compared to non-CSC using LC-MS/MS and confirmed those results using qRT-PCR and Western blotting . P02794 1 ( P02794 ) was identified as a candidate gene , which is involved in iron metabolism and iron depletion significantly decreased the self-renewal of CSCs . We further performed in silico analysis of altered genes in tumorsphere and identified a set of genes ( P06454 , P26447 , P06703 , TNXRD1 , P23219 , P35354 , P02533 , and P02794 ) , representing possible molecular targets , which in combination showed a promise to be used as prognostic markers for breast cancer . [ Meloxicam ( DB00814 ) : a review of its pharmacological and clinical profile ] . Meloxicam ( DB00814 ) is a new nonsteroidal anti-inflammatory drug ( NSAID ) derived from enolic acid , exhibiting selectivity for cyclooxygenase ( P36551 ) -2 over P23219 . Meloxicam has shown potent anti-inflammatory and analgesic activity together with low gastrointestinal toxicity in animal models . It is a potent inhibitor not only of acute exudation in adjuvant arthritis in the rat , but also of bone and cartilage destruction . The therapeutic range of meloxicam in the rat , with regard to inhibition of adjuvant arthritis , was several times greater than that of other NSAIDs . Meloxicam in therapeutic doses was found to have no effect on bleeding time or platelet aggregation in healthy volunteers . In clinical studies , meloxicam has shown reliable efficacy against rheumatoid arthritis , osteoarthritis , lumbago ( low back pain ) , scapulohumeral periarthritis , and neck-shoulder-arm syndrome with low gastrointestinal toxicity . Role of the JAK- P35610 pathway in protection against myocardial ischemia/reperfusion injury . The Janus kinase ( JAK ) -signal transducers and activators of transcription ( P35610 ) pathway is a stress-responsive mechanism that transduces signals from the cell surface to the nucleus , thereby modulating gene expression . Recent studies have demonstrated that myocardial ischemia and reperfusion induce rapid activation of this pathway . Although the functional consequences of this event remain to be elucidated , there is emerging evidence that JAK- P35610 signaling plays an important role in the development of the cardioprotected phenotype associated with ischemic preconditioning . Specifically , brief episodes of myocardial ischemia/reperfusion activate P23458 and O60674 , followed by recruitment of P42224 and P40763 , resulting in transcriptional upregulation of inducible nitric oxide synthase ( P35228 ) and cyclooxygenase-2 ( P35354 ) , which then mediate the infarct-sparing effects of the late phase of preconditioning . The present review focuses on this novel cardioprotective role of JAK- P35610 signaling and on its potential exploitation for developing therapeutic strategies aimed at limiting ischemia/reperfusion injury .

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interacts_with DB00159?

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Curcumin : getting back to the roots . The use of turmeric , derived from the root of the plant Curcuma longa , for treatment of different inflammatory diseases has been described in Ayurveda and in traditional Chinese medicine for thousands of years . The active component of turmeric responsible for this activity , curcumin , was identified almost two centuries ago . Modern science has revealed that curcumin mediates its effects by modulation of several important molecular targets , including transcription factors ( e.g. , NF-kappaB , AP-1 , Egr-1 , beta-catenin , and P37231 ) , enzymes ( e.g. , P35354 , 5- P28300 , P35228 , and hemeoxygenase-1 ) , cell cycle proteins ( e.g. , cyclin D1 and P38936 ) , cytokines ( e.g. , P01375 , IL-1 , P05231 , and chemokines ) , receptors ( e.g. , P00533 and P04626 ) , and cell surface adhesion molecules . Because it can modulate the expression of these targets , curcumin is now being used to treat cancer , arthritis , diabetes , Crohn 's disease , cardiovascular diseases , osteoporosis , Alzheimer 's disease , psoriasis , and other pathologies . Interestingly , 6-gingerol , a natural analog of curcumin derived from the root of ginger ( Zingiber officinalis ) , exhibits a biologic activity profile similar to that of curcumin . The efficacy , pharmacologic safety , and cost effectiveness of curcuminoids prompt us to " get back to our roots . " Nearly Complete Response of Brain Metastases from P04626 Overexpressing Breast Cancer with DB01259 and DB01101 after Whole Brain Irradiation . DB00072 treatment does not prevent intracranial seeding and is largely ineffective for established central nervous system metastasis in P04626 overexpressing breast cancer patients . Combination therapy of lapatinib and capecitabine may be an effective treatment option for brain metastasis of P04626 -positive breast cancer . We report a patient with breast cancer overexpressing HER-2 where brain metastases were successfully treated with radiation and a combination of lapatinib and capecitabine . P37840 A30P point-mutation generates age-dependent nigrostriatal deficiency in mice . Lewy bodies are mainly composed of alpha-synuclein ( P37840 ) and specific mutations in P37840 gene are related to familial forms of Parkinson 's disease ( PD ) . The purpose of our study was to generate a mouse line with A30P knock-in point mutation in P37840 gene and to test if a single point-mutation is able to turn otherwise normal P37840 into a toxic form . The behavioral profile of P37840 A30P mice was followed for 16 months . Generally , these mice are healthy and viable without any obvious abnormalities . Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function ( ink-test and beam walk ) . In other tests ( motility boxes , rotarod ) mice continuously performed normally . Moreover , P37840 A30P mice expressed the altered sensitivity to Q05940 inhibitor reserpine , possibly reflecting a functional deficiency of dopamine . Indeed , mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum , and reduced levels of dopamine in the mesolimbic system . The present study confirms that P37840 plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance . The generated mouse line has a potential to become a model for PD with comparable time course and phenotype . Characterization of the aggregation responses of camel platelets . BACKGROUND : Despite evidence of active hemostasis , camel platelets barely respond to common aggregating agents at standard doses used for human platelet aggregation . OBJECTIVES : The purpose of the study was to find out whether camel platelets can be activated by high doses or combinations of aggregation agonists , and to characterize the receptor that mediates the aggregation response to adenosine diphosphate ( ADP ) , the most potent agonist for camel platelets known so far . METHODS : Aggregation studies were performed with platelet-rich plasma ( PRP ) in response to multiple doses or combinations of ADP , epinephrine ( P08473 ) , collagen , and arachidonic acid ( AA ) . Aggregation responses to ADP were performed before and after the addition of the ADP receptor ( Q9H244 ) antagonist DB00758 . RESULTS : Camel platelets responded to ADP at doses higher than the standard dose for human platelets , and to combinations of P08473 and other agonists , while no aggregation was elicited with P08473 or AA alone . DB00758 blocked the ADP-induced aggregation responses in a dose-dependent fashion in vitro . CONCLUSIONS : Camel platelet aggregation can be activated by increasing the dose of some agonists such as ADP , but not AA or P08473 . Irreversible aggregation of camel platelets could also be triggered by a combination of P08473 and ADP , and collagen and AA . Inhibition with clopidogrel suggests that camel platelets express the ADP receptor , Q9H244 . Understanding platelet function in camels will add to the understanding of platelet function in health and disease . Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre- and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 ) -2 and epidermal growth factor receptor ( P00533 ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 ) was greater than 0.5 , seven ( 100 % , p=0.0515 ) and five ( 71.4 % , p=0.3742 ) patients showed positive immunoreactivity for P35354 and P00533 , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 and P00533 at pre-RT were associated with P30518 ( p=0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 , and six out of the eight patients had a P30518 greater than 0.5 ( p=0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 and P00533 . [ Anticoagulants of primary haemostasis ] . Inhibition of platelet function plays an important role in the treatment and secondary prevention of cardiovascular or cerebrovascular ischemic diseases . Established antiplatelet agents use different pharmacological targets for this role . Acetyl salicylic acid achieves a reduction of thromboxane A2 formation by inhibition of P23219 . DB00208 or clopidogrel are ADP- Q9H244 receptor antagonists . Tirofiban , abciximab or eptifibatid are used for the inhibition of the glycoprotein IIb/IIIa receptor which is activated at the surface of platelets preceding the final step of their aggregation . The mechanism of dipyridamole is based on the inhibition of adenosine uptake and of phosphodiesterase-5 . Efforts are made to improve antiplatelet therapy with the aim to find agents with favorable clinical outcome and lower bleeding risk . Current clinical studies focus on a new generation of ADP receptor antagonists ( prasugrel , cangrelor and ticagrelor ) as successors of ticlopidine and clopidogrel after coronary arterial interventions . Developments using platelet targets different from established drugs are thrombin receptor antagonists ( like SCH530348 ) or thromboxane receptor antagonists ( like S18886/terutroban ) in patients with cerebrovascular events . Results from recent experimental studies could lead to new strategies for antiplatelet therapy ( like inhibition of GP Ib receptor , GP VI receptor , platelet-leukocyte interaction , factor XII and others ) in the future . A novel mutation in P30518 causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6-month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor-2 gene ( P30518 ) located on chromosome Xq28 demonstrated a novel 5-base pair deletion ( c.962-966delACCCC ; g.1429-1433delACCCC ) leading to a shift of the reading frame ( p.Asn321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course . DB00159 inhibits prostaglandin D2 generation by inhibiting cyclo-oxygenase-2 in cultured human mast cells . BACKGROUND : DB00159 ( EPA ) is catalysed by cyclo-oxygenase ( P36551 ) , as is arachidonic acid , and is a competitive inhibitor of arachidonate metabolism . OBJECTIVES : We examined the effect of EPA on prostaglandin ( PG ) D2 generation in the cultured human mast cells with IgE-anti-IgE challenge incubation . METHODS : Cultured human mast cells were incubated with EPA ( 1 micromol/L ) for 20 h , then challenged with anti-IgE incubation after treatment with IgE . At the same time , P36551 inhibitors were tested to identify P23219 and P35354 activity . PGD2 synthetic activity was also assayed in a cell-free homogenate of cultured mast cells with P36551 inhibitors and EPA . DB11320 in the culture medium and in cells was assayed with the HPLC-fluorescent method . PGD2 and PGD3 were assayed with gas chromatography-mass spectrometry and the stable isotope dilution method . RESULTS : Although EPA incubation did not affect histamine release by cultured human mast cells in response to IgE-anti-IgE challenge incubation , it did decrease PGD2 generation by inhibiting the P35354 pathway . In contrast , in the cell-free homogenate of cultured human mast cells , EPA inhibited both P23219 and P35354 activities . CONCLUSION : Pre-incubation with EPA primarily affects the P35354 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE-anti-IgE challenge incubation . These findings suggest that P23219 and P35354 have different substrate flow systems in mast cells . They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti-inflammatory substrate in human mast cells . Identification of the amino acid sequence motif of alpha-synuclein responsible for macrophage activation . P37840 ( Syn ) is implicated in the pathogenesis of PD and related neurodegenerative disorders . Recent studies have also shown that alpha-synuclein can activate microglia and enhance dopaminergic neurodegeneration . The mechanisms of microglia activation by alpha-synuclein , however , are not well understood . In this study , we found that not only alpha-synuclein but also beta- and gamma-synucleins activated macrophages ( RAW 264.7 ) in vitro . Macrophages treated with synuclein proteins secreted P01375 in a dose-dependent manner . Synuclein family proteins also increased mRNA transcription of P35354 and P35228 . Two alpha-synuclein deletion mutants , SynDeltaNAC and Syn61-140 , activated macrophages , while deletion mutants Syn1-60 and Syn96-140 did not significantly activate them . Finally , we demonstrated that macrophage activation by alpha-synuclein was accompanied by phosphorylation of P29323 . These results suggest that synuclein family proteins can activate macrophages , and that macrophage activation needs both the N-terminal and C-terminal domains of alpha-synuclein , but not the central Q9C000 region . Sources contributing to the average extracellular concentration of dopamine in the nucleus accumbens . Mesolimbic dopamine neurons fire in both tonic and phasic modes resulting in detectable extracellular levels of dopamine in the nucleus accumbens ( NAc ) . In the past , different techniques have targeted dopamine levels in the NAc to establish a basal concentration . In this study , we used in vivo fast scan cyclic voltammetry ( FSCV ) in the NAc of awake , freely moving rats . The experiments were primarily designed to capture changes in dopamine caused by phasic firing - that is , the measurement of dopamine ' transients ' . These FSCV measurements revealed for the first time that spontaneous dopamine transients constitute a major component of extracellular dopamine levels in the NAc . A series of experiments were designed to probe regulation of extracellular dopamine . DB00281 was infused into the ventral tegmental area , the site of dopamine cell bodies , to arrest neuronal firing . While there was virtually no instantaneous change in dopamine concentration , longer sampling revealed a decrease in dopamine transients and a time-averaged decrease in the extracellular level . Dopamine transporter inhibition using intravenous GBR12909 injections increased extracellular dopamine levels changing both frequency and size of dopamine transients in the NAc . To further unmask the mechanics governing extracellular dopamine levels we used intravenous injection of the vesicular monoamine transporter ( Q05940 ) inhibitor , tetrabenazine , to deplete dopamine storage and increase cytoplasmic dopamine in the nerve terminals . DB04844 almost abolished phasic dopamine release but increased extracellular dopamine to ∼500 nM , presumably by inducing reverse transport by dopamine transporter ( Q01959 ) . Taken together , data presented here show that average extracellular dopamine in the NAc is low ( 20-30 nM ) and largely arises from phasic dopamine transients . Q13547 contributes to cell cycle and apoptosis . Histone deacetylases ( HDACs ) are generally thought to play important roles in human disease . However , little information is available concerning the specific functions of individual HDACs . We previously reported on transgenic mice that expressed human Q13547 and experienced steatosis and nuclear pleomorphism in their hepatic tissues . To find out if the over-expression of Q13547 contributes to the expression of genes related to the cell cycle , apoptosis , and lipid metabolism that eventually contribute to the pathological changes in the livers of the transgenic mice , the expression profiles of the related genes in liver tissues were determined by reverse transcription-polymerase chain reaction ( RT-PCR ) and Western blot analysis . The activated human Q13547 significantly induced the expression levels of mRNA for p53 , P37231 and Bak and reduced the P38936 expression level compared with the levels in control littermates . However , the protein levels of p53 and P37231 were significantly decreased . In conclusion , our results indicate that Q13547 can regulate gene expression at the mRNA and protein levels independently and that this may be a potential cytopathic factor for hepatic tissue in transgenic mice that over-express Q13547 . Effect of milk hydrolysates on inflammation markers and drug-induced transcriptional alterations in cell-based models . Nonsteroidal anti-inflammatory drugs ( NSAID ) are associated with gastrointestinal inflammation and subsequent damage to the intestinal tissue . Earlier studies in our laboratory have found that specific casein hydrolysates ( CH ) might be useful in the treatment of gastrointestinal wounds . The underlying mechanisms that support inflammation and wound healing are not completely understood , but transcriptional alterations may be used as markers for inflammation and wound healing . The bioactivity of 3 CH prepared by treatment of commercial casein with pepsin ( 60 min ) followed by corolase ( 0 , 10 , or 60 min ) were investigated in intestinal epithelial cells treated with the NSAID indomethacin . The bioactivity was evaluated as transcriptional alterations of transforming growth factor-β1 ( TGF-β1 ) , cyclooxygenase-2 ( P35354 ) , peroxisome proliferator-activated receptor-γ ( Q07869 -γ ) and nuclear factor κB ( NFκB ) by real-time PCR . Furthermore , the effect of CH on lipopolysaccharide-induced inflammation was evaluated in macrophages by measuring PG E(2) levels . Casein hydrolysates treated with corolase for 10 or 60 min after pepsin treatment downregulated transcription of TGF-β1 and NFκB ( P < 0.05 ) compared with the hydrolysate treated with pepsin only . Hydrolysate prepared by corolase treatment for 60 min after pepsin hydrolysis downregulated transcription of P35354 ( P < 0.05 ) compared with hydrolysate treated with corolase for only 10 min whereas transcription of Q07869 -γ was not affected ( P > 0.05 ) . Additionally , the hydrolysate prepared by pepsin treatment only ( 0 min corolase ) had a pro-inflammatory effect on macrophages via PG E(2) stimulation ( P < 0.05 ) . In conclusion , CH produced by a combination of pepsin and corolase treatments downregulated the transcription levels of TGF-β1 , P35354 , and NFκB . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . P62158 and troponin C : affinity chromatographic study of divalent cation requirements for troponin I inhibitory peptide ( residues 104-115 ) , mastoparan and fluphenazine binding . The different conformations induced by the binding of Mg2+ or Ca2+ to troponin C ( TnC ) and calmodulin ( P62158 ) results in the exposure of various interfaces with potential to bind target compounds . The interaction of TnC or P62158 with three affinity columns with ligands of either the synthetic peptide of troponin I ( TnI ) inhibitory region ( residues 104-115 ) , mastoparan ( a wasp venom peptide ) , or fluphenazine ( a phenothiazine drug ) were investigated in the presence of Mg2+ or Ca2+ . TnC and P62158 in the presence of either Ca2+ or Mg2+ bound to the TnI peptide 104-115 . The cation specificity for this interaction firmly establishes that the TnI inhibitory region binds to the high affinity sites of TnC ( most likely the N-terminal helix of site III ) and presumably the homologous region of P62158 . Mastoparan interacted strongly with both proteins in the presence of Ca2+ but , in the presence of Mg2+ , did not bind to TnC and only bound weakly to P62158 . DB00623 bound to TnC and P62158 only in the presence of Ca2+ . When the ligands interacted with either proteins there was an increase in cation affinity , such that TnC and P62158 were eluted from the TnI peptide or mastoparan affinity column with 0.1 M DB00974 compared with the 0.01 M DB00974 required to elute the proteins from the fluphenazine column . The interaction of these ligands with their receptor sites on TnC and P62158 require a specific and spatially correct alignment of hydrophobic and negatively charged residues on these proteins. ( ABSTRACT TRUNCATED AT 250 WORDS ) Discrete roles for peroxisome proliferator-activated receptor gamma and retinoid X receptor in recruiting nuclear receptor coactivators . P37231 ( PPARgamma ) plays a major role in adipogenesis . PPARgamma binds to DNA as a heterodimer with retinoid X receptor ( RXR ) , and PPARgamma-RXR can be activated by ligands specific for either receptor ; the presence of both ligands can result in a cooperative effect on the transactivation of target genes . How these ligands mediate transactivation , however , remains unclear . PPARgamma is known to interact with both the P52701 / Q15788 family of coactivators and the distinct , multisubunit coactivator complex called DRIP . A single DRIP subunit , Q15648 ( Q15648 , PBP ) , binds directly to PPARgamma . Here we report that PPARgamma and RXR selectively interacted with Q15648 and P52701 proteins in a ligand-dependent manner . At physiological concentrations , RXR-specific ligands only induced P52701 binding to RXR , and PPARgamma-specific ligands exclusively recruited Q15648 but not P52701 coactivators to PPARgamma . This selectivity was not observed in interaction assays off DNA , implying that the specificity of coactivator binding in response to ligand is strongly influenced by the allosteric effects of DNA-bound heterodimers . These coactivator-selective effects were also observed in transient-transfection assays in the presence of overexpressed P52701 or DRIP coactivators . The results suggest that the cooperative effects of PPARgamma- and RXR-specific ligands may occur at the level of selective coactivator recruitment . DB00159 modulates DB00091 -induced proinflammatory cytokine over-expression in osteoblastic cells in vitro . Several adverse outcomes are reported in subjects undergoing long term DB00091 ( DB00091 ) treatment . Severe osteopenia has been described in clinical and experimental reports , while beneficial effects of n-3 polyunsaturated fatty acids ( PUFAs ) on bone metabolism are recognized . In the present study we investigated the effects of n-3 versus n-6 PUFAs on osteoblastic cells treated with DB00091 , evaluating the expression of interleukin ( IL ) -1 & #223 ; , interleukin-6 ( P05231 ) , inducible nitric oxide synthase ( P35228 ) , and cyclooxygenase-2 ( P35354 ) in two different experimental protocols and the production of P05231 , IL-1 & #223 ; , and tumor necrosis factor alpha ( TNFalpha ) in cells challenged simultaneously with DB00091 and eicosapentaenoic acid ( EPA ) for 48h . IL-1 & #223 ; and P05231 up-regulation , induced by DB00091 , was counteracted by the addition of EPA in both protocols ; on the contrary , arachidonic acid ( AA ) magnified DB00091 the effects . P35354 and P35228 levels were not modified by DB00091 treatment . These in vitro results , that substantiate clinical reports of DB00091 -induced osteopenia , demonstrate a beneficial effect of EPA on DB00091 -altered cytokine profile , opening new perspectives in the non-pharmacological management of adverse outcomes in DB00091 -treated patients . Peroxisome-proliferator activator receptor-gamma activation decreases attachment of endometrial cells to peritoneal mesothelial cells in an in vitro model of the early endometriotic lesion . The aim of this study was to investigate whether peroxisome proliferator-activated receptor ( Q07869 ) -gamma activation has an effect on the attachment of endometrial cells to peritoneal mesothelial cells in a well-established in vitro model of the early endometriotic lesion . The endometrial epithelial cell line EM42 and mesothelial cell line LP9 were used for this study . EM42 cells , LP9 cells or both were treated with the P37231 agonist ciglitazone ( CTZ ) at varying concentrations ( 10 , 20 and 40 microM ) x 48 h with subsequent co-culture of EM42 and LP9 cells . The rate of EM42 attachment and invasion through LP9 cells was then assessed and compared with control ( EM42 and LP9 cells co-cultured without prior treatment with CTZ ) . Next , attachment of CTZ-treated and untreated EM42 cells to hyaluronic acid ( HA ) , a cell adhesion molecule ( P62158 ) on peritoneal mesothelial cells , were assessed . Although there was no difference in EM42 attachment when LP9 cells alone were treated with CTZ , treatment of EM42 cells with 40 microM CTZ decreased EM42 attachment to LP9 cells by 27 % ( P < 0.01 ) . Treatment of both EM42 and LP9 cells with 40 microM CTZ decreased EM42 attachment to LP9 by 37 % ( P < 0.01 ) . Treatment of EM42 cells with 40 microM CTZ decreased attachment to HA by 66 % ( P = 0.056 ) . CTZ did not decrease invasion of EM42 cells through the LP9 monolayer . CTZ may inhibit EM42 cell proliferation . In conclusion , CTZ significantly decreased EM42 attachment to LP9 cells and HA in an in vitro model of the early endometriotic lesion . Novel cinnamyl hydroxyamides and 2-aminoanilides as histone deacetylase inhibitors : apoptotic induction and cytodifferentiation activity . Four novel series of cinnamyl-containing histone deacetylase ( HDAC ) inhibitors 1-4 are described , containing hydroxamate ( 1 and 3 ) or 2-aminoanilide ( 2 and 4 ) derivatives . When screened against class I ( maize HD1-B and human Q13547 ) and class II ( maize HD1-A and human P56524 ) HDACs , most hydroxamates and 2-aminoanilides displayed potent and selective inhibition toward class I enzymes . Immunoblotting analyses performed in U937 leukemia cells generally revealed high acetyl-H3 and low acetyl-α-tubulin levels . Exceptions are compounds 3 f-i , 3 m-o , and 4 k , which showed higher tubulin acetylation than DB02546 . In U937 cells , cell-cycle blockade in either the G₂/M or G₁/S phase was observed with 1-4 . Five hydroxamates ( compounds 1 h-l ) effected a two- to greater than threefold greater percent apoptosis than DB02546 , and in the CD11c cytodifferentiation test some 2-aminoanilides belonging to both series 2 and 4 were more active than MS-275 . The highest-scoring derivatives in terms of apoptosis ( 1 k , 1 l ) or cytodifferentiation ( 2 c , 4 n ) also showed antiproliferative activity in U937 cells , thus representing valuable tools for study in other cancer contexts . Conditional ablation of mediator subunit MED1 ( MED1/ Q15648 ) gene in mouse liver attenuates glucocorticoid receptor agonist dexamethasone-induced hepatic steatosis . P04150 ( GR ) agonist dexamethasone ( DB00514 ) induces hepatic steatosis and enhances constitutive androstane receptor ( CAR ) expression in the liver . CAR is known to worsen hepatic injury in nonalcoholic hepatic steatosis . Because transcription coactivator MED1/ Q15648 gene is required for GR- and CAR-mediated transcriptional activation , we hypothesized that disruption of MED1/ Q15648 gene in liver cells would result in the attenuation of DB00514 -induced hepatic steatosis . Here we show that liver-specific disruption of MED1 gene ( MED1 ( delta Liv ) ) improves DB00514 -induced steatotic phenotype in the liver . In wild-type mice DB00514 induced severe hepatic steatosis and caused reduction in medium- and short-chain acyl- DB01992 dehydrogenases that are responsible for mitochondrial beta-oxidation . In contrast , DB00514 did not induce hepatic steatosis in mice conditionally null for hepatic MED1 , as it failed to inhibit fatty acid oxidation enzymes in the liver . MED1 ( delta Liv ) livers had lower levels of GR-regulated CAR mRNA compared to wild-type mouse livers . Microarray gene expression profiling showed that absence of MED1 affects the expression of the GR-regulated genes responsible for energy metabolism in the liver . These results establish that absence of MED1 in the liver diminishes DB00514 -induced hepatic steatosis by altering the GR- and CAR-dependent gene functions . Glucocorticoids enhance regeneration of murine olfactory epithelium . CONCLUSION : Glucocorticoid ( GC ) administration enhanced apoptotic changes in mature olfactory receptor neurons ( ORNs ) . GC administration may enhance regeneration of olfactory epithelium ( OE ) . OBJECTIVES : The mechanism underlying olfactory epithelial cells turnover involves apoptosis replaced by new ORNs . On regeneration of OE , we evaluated the apoptotic changes in OE . Our aim was to corroborate the enhancement of apoptosis of ORNs induced by GCs that are generally administered locally or systemically to patients with olfactory dysfunction . MATERIALS AND METHODS : For the in vitro study , we established cultured murine ORNs . DB00620 acetonide was added to culture supernatants . ORNs were then cultured for another 2 weeks . In the in vivo study , triamcinolone acetonide was administered to mice 5 or 10 times . The mice were dissected 3 days after the final injection , and the olfactory regions were removed and embedded in paraffin . All samples were examined by immunohistochemical staining and the TdT-mediated dUTP-biotin nick-end labeling ( TUNEL ) method . RESULTS : P04150 ( GR ) expression of cultured murine ORNs was observed among ORNs at the mature stage . Expression of GRs by murine OE was localized on mature ORNs and supporting cells . Administration of GC to both cultured ORNs and mice resulted in proportions of apoptotic cells that were significantly higher than those in the control groups . Omega-3 polyunsaturated fatty acids and inflammatory processes : nutrition or pharmacology ? DB00159 ( EPA ) and docosahexaenoic acid ( DB01708 ) are n-3 fatty acids found in oily fish and fish oil supplements . These fatty acids are able to inhibit partly a number of aspects of inflammation including leucocyte chemotaxis , adhesion molecule expression and leucocyte-endothelial adhesive interactions , production of eicosanoids like prostaglandins and leukotrienes from the n-6 fatty acid arachidonic acid , production of inflammatory cytokines and T cell reactivity . In parallel , EPA gives rise to eicosanoids that often have lower biological potency than those produced from arachidonioc acid and EPA and DB01708 give rise to anti-inflammatory and inflammation resolving resolvins and protectins . Mechanisms underlying the anti-inflammatory actions of n-3 fatty acids include altered cell membrane phospholipid fatty acid composition , disruption of lipid rafts , inhibition of activation of the pro-inflammatory transcription factor nuclear factor kappa B so reducing expression of inflammatory genes , activation of the anti-inflammatory transcription factor P37231 ( i.e. peroxisome proliferator activated receptor γ ) and binding to the G protein coupled receptor Q5NUL3 . These mechanisms are interlinked . In adult humans , an EPA plus DB01708 intake greater than 2 g day⁻¹ seems to be required to elicit anti-inflammatory actions , but few dose finding studies have been performed . Animal models demonstrate benefit from n-3 fatty acids in rheumatoid arthritis ( RA ) , inflammatory bowel disease ( Q9UKU7 ) and asthma . Clinical trials of fish oil in patients with RA demonstrate benefit supported by meta-analyses of the data . Clinical trails of fish oil in patients with Q9UKU7 and asthma are inconsistent with no overall clear evidence of efficacy . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . Functional changes in rheumatoid fibroblast-like synovial cells through activation of peroxisome proliferator-activated receptor gamma-mediated signalling pathway . P37231 ( PPARgamma ) is a ligand dependent transcriptional factor known to be a regulator of adipogenesis . Recent studies have also shown that stimulation of PPARgamma inhibits the transcriptional activities of other nuclear factors and down-regulates proinflammatory cytokine synthesis in T cells and monocytes . We examined , in the present study , the functional significance of PPARgamma expressed in fibroblast-like synovial cells ( FLS ) isolated from patients with rheumatoid arthritis ( RA ) . Incubation of FLS with a synthetic PPARgamma ligand , troglitazone , inhibited endogenous production of P01375 , P05231 and P10145 , as well as matrix metalloprotease-3 ( P08254 ) , without inducing apoptosis of the cells . The gelatinase activity of FLS culture media was also inhibited by troglitazone . Electrophoretic mobility shift assay ( EMSA ) showed a significant reduction in the DNA binding activity of NF-kappaB in troglitazone-treated FLS in response to P01375 or IL-1beta . Moreover , long-term cultivation of FLS with troglitazone resulted in morphological changes with marked lipid accumulation in these cells . Our results show a negative regulatory function for PPARgamma on cytokine and MMP production together with inhibition of cytokine-mediated inflammatory responses in rheumatoid synovial cells . Our results also suggest that FLS could differentiate into adipocyte-like cells in the presence of proper stimulatory signals including PPARgamma .

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Inhibition of mutant P15056 splice variant signaling by next-generation , selective RAF inhibitors . DB08881 and dabrafenib block MEK- P27361 /2 signaling and cause tumor regression in the majority of advanced-stage P15056 (V600E) melanoma patients ; however , acquired resistance and paradoxical signaling have driven efforts for more potent and selective RAF inhibitors . Next-generation RAF inhibitors , such as PLX7904 ( PB04 ) , effectively inhibit RAF signaling in P15056 (V600E) melanoma cells without paradoxical effects in wild-type cells . Furthermore , PLX7904 blocks the growth of vemurafenib-resistant P15056 (V600E) cells that express mutant P01111 . Acquired resistance to vemurafenib and dabrafenib is also frequently driven by expression of mutation P15056 splice variants ; thus , we tested the effects of PLX7904 and its clinical analog , PLX8394 ( PB03 ) , in P15056 (V600E) splice variant-mediated vemurafenib-resistant cells . We show that paradox-breaker RAF inhibitors potently block MEK- P27361 /2 signaling , P55008 /S cell cycle events , survival and growth of vemurafenib/PLX4720-resistant cells harboring distinct P15056 (V600E) splice variants . These data support the further investigation of paradox-breaker RAF inhibitors as a second-line treatment option for patients failing on vemurafenib or dabrafenib . Drug-induced activation of SREBP-controlled lipogenic gene expression in CNS-related cell lines : marked differences between various antipsychotic drugs . BACKGROUND : The etiology of schizophrenia is unknown , but neurodevelopmental disturbances , myelin- and oligodendrocyte abnormalities and synaptic dysfunction have been suggested as pathophysiological factors in this severe psychiatric disorder . DB04540 is an essential component of myelin and has proved important for synapse formation . Recently , we demonstrated that the antipsychotic drugs clozapine and haloperidol stimulate lipogenic gene expression in cultured glioma cells through activation of the sterol regulatory element-binding protein ( SREBP ) transcription factors . We here compare the action of chlorpromazine , haloperidol , clozapine , olanzapine , risperidone and ziprasidone on SREBP activation and SREBP-controlled gene expression ( ACAT2 , P04035 , Q01581 , P14324 , O75845 , Q9UBM7 , P01130 , P49327 and SCD1 ) in four CNS-relevant human cell lines . RESULTS : There were marked differences in the ability of the antipsychotic drugs to activate the expression of SREBP target genes , with clozapine and chlorpromazine as the most potent stimulators in a context of therapeutically relevant concentrations . Glial-like cells ( GaMg glioma and CCF-STTG1 astrocytoma cell lines ) displayed more pronounced drug-induced SREBP activation compared to the response in Q9UL51 human cortical neurons and SH-SY5Y neuroblastoma cells , indicating that antipsychotic-induced activation of lipogenesis is most prominent in glial cells . CONCLUSION : Our present data show a marked variation in the ability of different antipsychotics to induce SREBP-controlled transcriptional activation of lipogenesis in cultured human CNS-relevant cells . We propose that this effect could be relevant for the therapeutic efficacy of some antipsychotic drugs . Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 and Tissue P00747 Activator in Occluded Arteries . Human and mouse trace amine-associated receptor 1 have distinct pharmacology towards endogenous monoamines and imidazoline receptor ligands . TAARs ( trace amine-associated receptors ) are G-protein-coupled receptors that respond to low abundance , endogenous amines such as tyramine and tryptamine , and represent potential targets for neuropsychiatric diseases . However , some members of this receptor subfamily either have no ligand identified or remain difficult to express and characterize using recombinant systems . In the present paper we report the successful expression of human and mouse Q96RJ0 , and the characterization of their responses to various natural and synthetic agonists . In P29320 ( human embryonic kidney ) -293/CRE-bla cells , mouse Q96RJ0 showed a robust response to trace amines as measured using either a DB02527 assay or a beta-lactamase reporter assay , whereas human Q96RJ0 showed a weaker , but still measurable , response . When certain fragments of human Q96RJ0 were replaced with the corresponding regions of mouse Q96RJ0 , the chimaeric receptor showed a much stronger response in DB02527 production . Examination of a series of agonists on these receptors revealed that the human and the chimaeric receptor are almost identical in pharmacology , but distinct from the mouse receptor . We also screened small libraries of pharmacologically active agents on Q96RJ0 and identified a series of synthetic agonists , some of which are also ligands of the enigmatic imidazoline receptor . The findings of the present study not only shed light on the pharmacological species difference of Q96RJ0 , but also raise new possibilities about the mechanism of some of the imidazoline-related agents . De novo mutations in synaptic transmission genes including Q05193 cause epileptic encephalopathies . Emerging evidence indicates that epileptic encephalopathies are genetically highly heterogeneous , underscoring the need for large cohorts of well-characterized individuals to further define the genetic landscape . Through a collaboration between two consortia ( EuroEPINOMICS and Epi4K/EPGP ) , we analyzed exome-sequencing data of 356 trios with the " classical " epileptic encephalopathies , infantile spasms and Lennox Gastaut syndrome , including 264 trios previously analyzed by the Epi4K/EPGP consortium . In this expanded cohort , we find 429 de novo mutations , including de novo mutations in Q05193 in five individuals and de novo mutations in O75899 , P49327 , and Q15413 in two individuals each . Unlike previous studies , this cohort is sufficiently large to show a significant excess of de novo mutations in epileptic encephalopathy probands compared to the general population using a likelihood analysis ( p = 8.2 × 10(-4) ) , supporting a prominent role for de novo mutations in epileptic encephalopathies . We bring statistical evidence that mutations in Q05193 cause epileptic encephalopathy , find suggestive evidence for a role of three additional genes , and show that at least 12 % of analyzed individuals have an identifiable causal de novo mutation . Strikingly , 75 % of mutations in these probands are predicted to disrupt a protein involved in regulating synaptic transmission , and there is a significant enrichment of de novo mutations in genes in this pathway in the entire cohort as well . These findings emphasize an important role for synaptic dysregulation in epileptic encephalopathies , above and beyond that caused by ion channel dysfunction . Molecular identification of the human O75899 : cell surface expression and coupling to adenylyl cyclase in the absence of Q9UBS5 . We have identified a gene encoding a GABAB receptor , the human O75899 , located on chromosome 9q22.1 , that is distinct from the recently reported rat Q9UBS5 . O75899 structurally resembles Q9UBS5 ( 35 % identity ) , having seven transmembrane domains and a large extracellular region , but differs in having a longer carboxy-terminal tail . O75899 is localized to the cell surface in transfected COS cells , and negatively couples to adenylyl cyclase in response to GABA , baclofen , and 3-aminopropyl(methyl)phosphinic acid in CHO cells lacking Q9UBS5 . DB00181 action is inhibited by the GABABR antagonist , 2-hydroxysaclofen . The human O75899 and Q9UBS5 genes are differentially expressed in the nervous system , with the greatest difference being detected in the striatum in which Q9UBS5 but not O75899 mRNA transcripts are detected . O75899 and Q9UBS5 mRNAs are also coexpressed in various brain regions such as the Purkinje cell layer of the cerebellum . Identification of a functional homomeric O75899 coupled to adenylyl cyclase suggests that the complexity of GABAB pharmacological data is at least in part due to the presence of more than one receptor and opens avenues for future research leading to an understanding of metabotropic GABA receptor signal transduction mechanisms . Encapsulation of viral vectors for gene therapy applications . In gene therapy , a number of viruses are currently being used as vectors to provide transient expression of therapeutic proteins . A drawback of using free virus is that it gives a potent immune response , which reduces gene transfer and limits re-administration . An alternative delivery system is to encapsulate the virus in poly(lactide-co-glycolide) ( P00747 ) microspheres prior to administration . A recombinant adenovirus ( Ad ) expressing green fluorescent protein ( GFP ) was used to test the transduction efficiency of Ad encapsulated in microspheres on target cells . The number of infected cells that expressed GFP was measured by flow cytometry . It was demonstrated that encapsulated viral vectors could successfully transduce target cells with encapsulation efficiencies up to 23 % and that the level of transduction could be controlled by varying both the quantity of microspheres and the amount of Ad in the microspheres . High transduction efficiencies and its recognized biocompatibility make P00747 -encapsulated Ad an attractive alternative to the use of free virus in gene therapy applications . The infectivity of Ad was found to be significantly influenced by the processing conditions and changes in environmental factors . Free Ad and encapsulated Ad were able to infect both E1 complimenting cells ( P29320 293 ) and non-complimenting cells ( A549 ) , with the viral expression in P29320 293 cells being 2.1 times greater than for A549 cells . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . A complex of synaptic adhesion molecule Q9BY67 , a molecule related to autism spectrum disorder , with O75970 in the cerebellum . Mutations in the synaptic adhesion protein Q9BY67 ( RA175/SynCAM1 ) are associated with autism spectrum disorder ( P51689 ) , a neurodevelopmental disorder of uncertain molecular origin . Cadm1-knock out ( KO ) mice exhibit smaller cerebella with decreased number of synapse of Purkinje cells and some P51689 -like symptoms , including impaired ultrasonic vocalization . In this study , we examined the alteration of the Cadm1 synaptic complex in the mouse cerebellum at post-natal stages . The C-terminal peptide of Cadm1 associated with Mupp1 at P78352 /Dlg/ZO-1 ( PDZ ) (1-5) , a scaffold protein containing 13 PDZ domains , which interacted with gamma-aminobutyric acid type B receptor (GABBR)2 at PDZ13 , but not with P78352 . The O75899 was detected in a set of proteins interacting with Cadm1 C-terminal . Cadm1 colocalized with Mupp1 and O75899 on the dendrites of Purkinje cells in the molecular layers of the developing cerebellum and on the dendrites of hippocampal neurons cultured in vitro . These observations suggest that the Cadm1 synaptic receptor complex , including Mupp1- O75899 , is located on the dendrites of Purkinje cells . The amount of O75899 protein , but not mRNA , was increased in the cerebella of Cadm1 KO mice , suggesting that lack of Cadm1 does not affect transcription of O75899 , but may stabilize the Mupp1- O75899 complex ; the Mupp1- O75899 interaction may be stabilized by conformational change in Mupp1 or association with other adhesion molecules and by anchorage to the post-synaptic membrane . Up-regulation of O75899 in the cerebellum in the absence of Q9BY67 may be associated with P51689 pathogenesis . A novel mechanism of autophagic cell death in dystrophic muscle regulated by Q99572 receptor large-pore formation and HSP90 . Q99572 is an DB00171 -gated ion channel , which can also exhibit an open state with a considerably wider permeation . However , the functional significance of the movement of molecules through the large pore ( LP ) and the intracellular signaling events involved are not known . Here , analyzing the consequences of Q99572 activation in primary myoblasts and myotubes from the Dmd(mdx) mouse model of Duchenne muscular dystrophy , we found DB00171 -induced Q99572 -dependent autophagic flux , leading to P42574 - P55210 -independent cell death . Q99572 -evoked autophagy was triggered by LP formation but not Ca(2+) influx or P28482 - P27361 phosphorylation , 2 canonical Q99572 -evoked signals . Phosphoproteomics , protein expression inference and signaling pathway prediction analysis of Q99572 signaling mediators pointed to P54652 and HSP90 proteins . Indeed , specific HSP90 inhibitors prevented LP formation , LC3-II accumulation , and cell death in myoblasts and myotubes but not in macrophages . Pharmacological blockade or genetic ablation of p2rx7 also proved protective against DB00171 -induced death of muscle cells , as did inhibition of autophagy with 3-MA . The functional significance of the Q99572 LP is one of the great unknowns of purinergic signaling . Our data demonstrate a novel outcome -- autophagy -- and show that molecules entering through the LP can be targeted to phagophores . Moreover , we show that in muscles but not in macrophages , autophagy is needed for the formation of this LP . Given that Q99572 -dependent LP and HSP90 are critically interacting in the DB00171 -evoked autophagic death of dystrophic muscles , treatments targeting this axis could be of therapeutic benefit in this debilitating and incurable form of muscular dystrophy . P35367 antagonist cetirizine impairs working memory processing speed , but not episodic memory . BACKGROUND AND PURPOSE : The histaminergic neurotransmitter system is currently under investigation as a target for drug treatment of cognitive deficits in clinical disorders . The therapeutic potential of new drugs may initially be screened using a model of histaminergic dysfunction , for example , as associated with the use of centrally active antihistamines . Of the selective second generation antihistamines , cetirizine has been found to have central nervous system effects . The aim of the present study was to determine whether cetirizine can be used as a tool to model cognitive deficits associated with histaminergic hypofunction . EXPERIMENTAL APPROACH : The study was conducted according to a three-way , double-blind , cross-over design . Treatments were single oral doses of cetirizine 10 and 20 mg and placebo . Effects on cognition were assessed using tests of word learning , memory scanning , vigilance , divided attention , tracking and visual information processing speed . KEY RESULTS : DB00341 10 mg impaired tracking performance and both doses impaired memory scanning speed . None of the other measures indicated impaired performance . CONCLUSION AND IMPLICATIONS : DB00341 affects information processing speed , but these effects were not sufficient to serve as a model for cognitive deficits in clinical disorders . [ DB00707 sodium ( Photofrin-II ) ] . DB00707 sodium ( DB00707 ) is a photosensitizer which distributes selectively to tumor tissues , and causes tumor cell death by combination with light irradiation . Photodynamic therapy ( PDT ) by combination of porfimer sodium and laser was developed as a new cancer therapy . Tumor selectivity of porfimer sodium are based on the following reasons ; 1 ) high affinity for lipoprotein , especially , low density lipoprotein ( LDL ) , 2 ) elevation of P01130 activity in cancer tissue , and 3 ) lack or imcompleteness of lymphatic system in cancer tissue . DB00707 sodium is activated by laser irradiation at 630 nm , which can reacts with tissue oxygen to produce highly reactive excited siglet oxygen ( 1O2 ) . This highly reactive molecule is subsequently capable of killing tumor cells through oxidation of cellular component like mitochondrial enzymes . In addition , this highly reactive intermediate causes destruction of the tumor capillaries , which accelerates tumor cell death . The growth suppression or lethal damage to tumor cells by PDT of porfimer sodium and excimer dye laser were observed in experimental tumor models . In human clinical trials , the rates of complete response ( CR ) for roentgenographically occult lung cancer , stage I lung cancer , superficial esophageal cancer , superficial gastric cancer and carcinoma in situ or dysplasia of the cervix were 84.8 % , 50.0 % , 90.0 % , 87.5 % and 94.4 % , respectively . The major side effects were cutaneous symptoms e.g. photosensitivity , pigmentation , increasing GOT , GPT but these symptoms were not severe . PDT using porfimer sodium and excimer dye laser must be clinically useful for the treatment of inoperable early cancer or conservation of organ functions . Augmentation of methamphetamine-induced behaviors in transgenic mice lacking the trace amine-associated receptor 1 . The trace amine-associated receptor 1 ( Q96RJ0 ) is a G protein-coupled receptor that is functionally activated by amphetamine-based psychostimulants , including amphetamine , methamphetamine and DB01454 . Previous studies have shown that in transgenic mice lacking the Q96RJ0 gene ( Q96RJ0 knockout ; KO ) a single injection of amphetamine can produce enhanced behavioral responses compared to responses evoked in wild-type ( WT ) mice . Further , the psychostimulant effects of cocaine can be diminished by selective activation of Q96RJ0 . These findings suggest that Q96RJ0 might be implicated in the rewarding properties of psychostimulants . To investigate the role of Q96RJ0 in the rewarding effects of drugs of abuse , the psychomotor stimulating effects of amphetamine and methamphetamine and the conditioned rewarding effects of methamphetamine and morphine were compared between WT and Q96RJ0 KO mice . In locomotor activity studies , both single and repeated exposure to DB01576 or methamphetamine generated significantly higher levels of total distance traveled in Q96RJ0 KO mice compared to WT mice . In conditioned place preference ( CPP ) studies , Q96RJ0 KO mice acquired methamphetamine-induced CPP earlier than WT mice and retained CPP longer during extinction training . In morphine-induced CPP , both WT and KO genotypes displayed similar levels of CPP . Results from locomotor activity studies suggest that Q96RJ0 may have a modulatory role in the behavioral sensitization to amphetamine-based psychostimulants . That methamphetamine-but not morphine-induced CPP was augmented in Q96RJ0 KO mice suggests a selective role of Q96RJ0 in the conditioned reinforcing effects of methamphetamine . Collectively , these findings provide support for a regulatory role of Q96RJ0 in methamphetamine signaling . Small interfering RNA knocks down the molecular target of alendronate , farnesyl pyrophosphate synthase , in osteoclast and osteoblast cultures . P14324 ( FPPS ) , an enzyme in the mevalonate pathway , is the inhibition target of alendronate , a potent FDA-approved nitrogen-containing bisphosphonate ( N-BP ) drug , at the molecular level . DB00630 not only inhibits osteoclasts but also has been reported to positively affect osteoblasts . This study assesses the knockdown effects of siRNA targeting FPPS compared with alendronate in both osteoclast and osteoblast cultures . Primary murine bone marrow cell-induced osteoclasts and the preosteoblast MC3T3-E1 cell line were used to assess effects of anti-FPPS siRNA compared with alendronate . Results show that both FPPS mRNA message and protein knockdown in serum-based culture is correlated with reduced osteoclast viability . FPPS siRNA is more potent than 10 μM alendronate , but less potent than 50 μM alendronate on reducing osteoclast viability . Despite FPPS knockdown , no significant changes were observed in osteoblast proliferation . FPPS knockdown promotes osteoblast differentiation significantly but not cell mineral deposition . However , compared with 50 μM alendronate dosing , FPPS siRNA does not exhibit cytotoxic effects on osteoblasts while producing significant effects on ostoblast differentiation . Both siRNA and alendronate at tested concentrations do not have significant effects on cultured osteoblast mineralization . Overall , results indicate that siRNA against FPPS could be useful for selectively inhibiting osteoclast-mediated bone resorption and improving bone mass maintenance by influencing both osteoclasts and osteoblasts in distinct ways . DB11320 contributes to tissue remodeling via periostin expression . DB11320 is thought to have a critical role in the synthesis of extracellular matrix in skin and may be involved in tissue remodeling of allergic diseases . Recent studies revealed that periostin , a matricelluar protein , contributed to tissue remodeling ; however , a link between periostin and histamine remains unproven . We investigated whether periostin was involved in histamine-induced collagen production . Cultured dermal fibroblasts derived from wild-type ( WT ) or periostin knockout ( PN(-/-) ) mice were stimulated with histamine , and then collagen and periostin production was evaluated . DB11320 induced collagen gene expression in WT fibroblasts in the late phase but not in the early phase , whereas no effect on collagen expression was observed in histamine-stimulated PN(-/-) fibroblasts . In WT fibroblasts , histamine directly induced periostin expression in a dose-dependent manner , and an H1 receptor antagonist blocked both periostin and collagen expression . DB11320 activated extracellular signal-regulated kinase 1/2 ( P27361 /2 ) through the H1 receptor . Q15063 induction was inhibited by either H1 antagonist or P27361 /2 inhibitor treatment in vitro and was attenuated in P35367 (-/-) mice . Elevated expression of periostin was found in lesional skin from atopic dermatitis patients . These results suggest that histamine mediates periostin induction and collagen production through activation of the H1 receptor-mediated P27361 /2 pathway ; furthermore , histamine may accelerate the chronicity of atopic dermatitis . Type B gamma-aminobutyric acid receptors modulate the function of the extracellular Ca2+-sensing receptor and cell differentiation in murine growth plate chondrocytes . Extracellular calcium-sensing receptors ( CaRs ) and metabotropic or type B gamma-aminobutyric acid receptors ( GABA-B-Rs ) , two closely related members of family C of the G protein-coupled receptor superfamily , dimerize in the formation of signaling and membrane-anchored receptor complexes . We tested whether CaRs and two GABA-B-R subunits ( Q96GN5 and R2 ) are expressed in mouse growth plate chondrocytes ( GPCs ) by PCR and immunocytochemistry and whether interactions between these receptors influence the expression and function of the CaR and extracellular Ca(2+)-mediated cell differentiation . Both CaRs and the Q9UBS5 and -R2 were expressed in the same zones of the growth plate and extensively colocalized in intracellular compartments and on the membranes of cultured GPCs . The Q9UBS5 co-immunoprecipitated with the CaR , confirming a physical interaction between the two receptors in GPCs . In vitro knockout of Q9UBS5 genes , using a Cre-lox recombination strategy , blunted the ability of high extracellular Ca(2+) concentration to activate phospholipase C and P27361 /2 , suppressed cell proliferation , and enhanced apoptosis in cultured GPCs . In GPCs , in which the Q9UBS5 was acutely knocked down , there was reduced expression of early chondrocyte markers , aggrecan and type II collagen , and increased expression of the late differentiation markers , type X collagen and osteopontin . These results support the idea that physical interactions between CaRs and GABA-B-R1s modulate the growth and differentiation of GPCs , potentially by altering the function of CaRs . Localization of apolipoprotein E receptor 2 to caveolae in the plasma membrane . The P01130 ( LDL-R ) promotes the specific endocytosis and lysosomal delivery of extracellular lipoprotein ligands via clathrin-coated pits . It was widely assumed that other closely related members of the LDL-R gene family would have similar functions , but recent experimental evidence has revealed that one such protein , apolipoprotein E receptor 2 ( apoER2 ) , has a critical role as an " outside-in " signal transducer in the brain . ApoER2 signaling appears to require interaction between its cytoplasmic domain and adapter molecules such as Dab1 , JIP 1 and JIP 2 , and P78352 . Many of the receptors for other signaling pathways affected by such adapter molecules are compartmentalized into specialized microdomains within the plasma membrane termed caveolae . Here , we show that apoER2 , but not LDL-R , is localized to caveolae , supporting the concept that its physiological role is in cell signaling , rather than in endocytosing ligands . Immunocytochemical localization of Q9UBS5 receptor subunits in the basolateral amygdala . Gamma-aminobutyric acid B ( GABAB ) receptors ( GBRs ) are G-protein-coupled receptors that mediate a slow , prolonged form of inhibition in the basolateral amygdala ( P00519 ) and other brain areas . Recent studies indicate that this receptor is a heterodimer consisting of Q9UBS5 ( GBR1 ) and O75899 subunits . In the present investigation , antibodies to the Q9UBS5 subunit were used to study the neuronal localization of GBRs in the rat P00519 . GBR immunoreactivity was mainly found in spine-sparse interneurons and astrocytes at the light microscopic level . Very few pyramidal neurons exhibited perikaryal staining . Dual-labeling immunofluorescence analysis indicated that each of the four main subpopulations of interneurons exhibited GBR immunoreactivity . Virtually 100 % of large CCK+ neurons in the basolateral and lateral nuclei were GBR+ . In the basolateral nucleus 72 % of somatostatin ( Q8TE85 ) , 73 % of parvalbumin ( PV ) and 25 % of P01282 positive interneurons were GBR+ . In the lateral nucleus 50 % of somatostatin , 30 % of parvalbumin and 27 % of P01282 positive interneurons were GBR+ . Electron microscopic ( EM ) analysis revealed that most of the light neuropil staining seen at the light microscopic level was due to the staining of dendritic shafts and spines , most of which probably belonged to spiny pyramidal cells . Very few axon terminals ( Ats ) were GBR+ . In summary , this investigation demonstrates that the distal dendrites of pyramidal cells , and varying percentages of each of the four main subpopulations of interneurons in the P00519 , express GBRs . Because previous studies suggest that GBR-mediated inhibition modulates DB01221 -dependent EPSPs in the P00519 , these receptors may play an important role in neuronal plasticity related to emotional learning . Flow cytometric analysis of mammalian glial cultures treated with methotrexate . DB00563 ( MTX ) is an antineoplastic drug that acts by competitive inhibition of the enzyme dihydrofolate reductase ( P00374 ) . MTX treatment of cultured cell lines leads to the emergence of resistant cell populations . Studies using stepwise selection procedures have demonstrated that MTX resistance conferred by overproduction of P00374 can be caused by P00374 gene amplification . We examined the effect of MTX on cells whose origin more closely approximates the in vivo condition by developing a culture system using dissociated brain tissue from 17-19 day old mouse embryos . At the first passage , cultures were divided into control and MTX groups . Cells were treated with the same or successively higher concentrations of MTX at each passage over a 3-4 month period . The first passage eliminated neurons and left a glial culture comprised of approximately 90 % astrocytes . We used the Fluorescence Activated Cell Sorter in conjunction with fluorescent dyes to measure P00374 content , DNA content , size , and viability of glial cells following MTX treatment . MTX-treated cells divided but grew more slowly and were larger than untreated cells . Stepwise selection in 30/60/90 nM or 60/120 nM MTX resulted in significant two- to threefold increases in fluorescence , and hence P00374 levels . Slot hybridizations assays demonstrated a threefold increase in P00374 gene copy number in the DNA from the 30/60/90 cultures . Thus , our findings were consistent with the results obtained from somatic cell lines , and lend support to the hypothesis that gene amplification may be a common mechanism for the acquisition of resistance in many types of cells . They also indicate that glial cells may be a specific target for cytotoxic effects of MTX on the central nervous system . DB01098 improves endothelial function in db/db mice : role of angiotensin II type 1 receptors and oxidative stress . BACKGROUND AND PURPOSE : P04035 inhibitors , statins , with lipid-reducing properties combat against atherosclerosis and diabetes . The favourable modulation of endothelial function may play a significant role in this effect . The present study aimed to investigate the cellular mechanisms responsible for the therapeutic benefits of rosuvastatin in ameliorating diabetes-associated endothelial dysfunction . EXPERIMENTAL APPROACH : Twelve-week-old db/db diabetic mice were treated with rosuvastatin at 20 mg·kg⁻¹ ·day⁻¹ p.o.for 6 weeks . Isometric force was measured in isolated aortae and renal arteries . Protein expressions including angiotensin II type 1 receptor ( AT₁R ) , Q9NPH5 , O75935 (phox) , p67(phox) , Rac-1 , nitrotyrosine , phospho- P27361 /2 and phospho-p38 were determined by Western blotting , while reactive oxygen species ( ROS ) accumulation in the vascular wall was evaluated by dihydroethidium fluorescence and lucigenin assay . KEY RESULTS : DB01098 treatment of db/db mice reversed the impaired ACh-induced endothelium-dependent dilatations in both renal arteries and aortae and prevented the exaggerated contractions to angiotensin II and phenylephrine in db/db mouse renal arteries and aortae . DB01098 reduced the elevated expressions of AT₁R , O75935 (phox) and p67(phox) , Q9NPH5 , Rac1 , nitrotyrosine and phosphorylation of P27361 /2 and p38 MAPK and inhibited ROS production in aortae from db/db mice . CONCLUSIONS AND IMPLICATIONS : The vasoprotective effects of rosuvastatin are attributed to an increase in NO bioavailability , which is probably achieved by its inhibition of ROS production from the AT₁R-NAD(P)H oxidase cascade . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . Complex formation with the Type B gamma-aminobutyric acid receptor affects the expression and signal transduction of the extracellular calcium-sensing receptor . Studies with P29320 -293 cells and neurons . We co-immunoprecipitated the Ca(2+)-sensing receptor ( CaR ) and type B gamma-aminobutyric acid receptor ( GABA-B-R ) from human embryonic kidney ( P29320 ) -293 cells expressing these receptors and from brain lysates where both receptors are present . CaRs extensively co-localized with the two subunits of the GABA-B-R ( Q96GN5 and R2 ) in P29320 -293 cell membranes and intracellular organelles . Coexpressing CaRs and GABA-B-R1s in P29320 -293 cells suppressed the total cellular and cell surface expression of CaRs and inhibited phospholipase C activation in response to high extracellular [ Ca(2+) ] ( [Ca(2+)](e) ) . In contrast , coexpressing CaRs and GABA-B-R2s enhanced CaR expression and signaling responses to raising [Ca(2+)](e) . The latter effects of the O75899 on the CaR were blunted by coexpressing the Q9UBS5 . Coexpressing the CaR with Q9UBS5 or R2 enhanced the total cellular and cell surface expression of the Q9UBS5 or R2 , respectively . Studies with truncated CaRs indicated that the N-terminal extracellular domain of the CaR participated in the interaction of the CaR with the Q9UBS5 and R2 . In cultured mouse hippocampal neurons , CaRs co-localized with the Q9UBS5 and R2 . CaRs and GABA-B-R1s also co-immunoprecipitated from brain lysates . The expression of the CaR was increased in lysates from Q9UBS5 knock-out mouse brains and in cultured hippocampal neurons with their Q9UBS5 genes deleted in vitro . Thus , CaRs and GABA-B-R subunits can form heteromeric complexes in cells , and their interactions affect cell surface expression and signaling of CaR , which may contribute to extracellular Ca(2+)-dependent receptor activation in target tissues .

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[ P01308 resistance and hyperlipidemia in the elderly ] . To elucidate the relationship between hyperlipidemias and insulin resistance in the elderly , we conducted three studies : 1 ) determination of the prevalence of hyperlipidemias in elderly subjects with impaired glucose tolerance or non-insulin dependent diabetes mellitus , 2 ) measurement of plasma glucose and insulin levels in patients with phyerlipidemias and atherosclerotic vascular disease , and 3 ) computation of correlation between levels of substances involved in coagulation and fibrinolysis ( F-VII , F-X , and P05121 ) and levels of triglycerides and insulin in serum in hyperlipidemic patients with atherosclerotic vascular disease . The prevalence of hypertriglyceridemia was 4 % in non-diabetic control subjects , 38 % in subjects with impaired glucose tolerance , 22 % in those with diabetes , and 29 % in those with both conditions . Levels of glucose and insulin in plasma were measured after oral intake of 75 g of glucose . The insulin response was greater in the group with hypertriglyceridemia than in the group with normal triglyceride levels , although the glucose responses did not differ between the groups . The activities and levels of F-VII , F-X , and P05121 correlated with triglycerides in serum and also with fasting insulin levels in hyperlipidemic patients with atherosclerotic vascular disease . We conclude that hypertriglyceridemia plays an important role in the development of atherothrombotic vascular disease ; it accompanies elevation of coagulation and antifibrinolytic activities in elderly patients with insulin resistance . P01308 resistance determines phagocytic nicotinamide adenine dinucleotide phosphate oxidase overactivation in metabolic syndrome patients . OBJECTIVE : Metabolic syndrome ( MetS ) is associated with insulin resistance and increases the cardiovascular risk . Oxidative stress constitutes a potential mechanism that links insulin resistance and cardiovascular disease . The aim of this study was to analyze the relationship of NADPH oxidase activation with insulin resistance , and the effect of this interaction on the cardiovascular risk in MetS patients . METHODS : NADPH oxidase-dependent superoxide production and expression was evaluated by luminescence and western blot , respectively , in peripheral blood mononuclear cells obtained from 125 patients with MetS . P01308 resistance was defined by the homeostasis model assessment index . P14780 was quantified by enzyme-linked immunosorbent assay in plasma samples . To ascertain the mechanisms involved in vivo , we performed in-vitro experiments in cultured macrophages . RESULTS : Fifty-six percent of patients with MetS showed insulin resistance . Plasma matrix metalloproteinase-9 levels were higher ( P < 0.05 ) in insulin-resistant patients than in patients with insulin sensitivity . NADPH oxidase-dependent superoxide production was augmented ( P < 0.05 ) in insulin-resistant patients with respect to insulin-sensitive patients . The interaction between insulin resistance and abnormally high NADPH oxidase-mediated superoxide production was associated with the highest matrix metalloproteinase-9 values . Increased NADPH oxidase-dependent superoxide production was significantly associated with higher NADPH oxidase P13498 expression in insulin-resistant than in insulin-sensitive patients . Interestingly , insulin upregulated P13498 in peripheral blood mononuclear cells and in murine macrophages . CONCLUSION : P01308 resistance is associated with phagocytic NADPH oxidase activation . This association results in the highest cardiovascular risk in MetS patients . Systems pharmacology assessment of the 5-fluorouracil pathway . AIM : To assess the impact of the 5-fluorouracil ( DB00544 ) drug-pathway genes on cytotoxicity , and determine whether loss-of-function analyses coupled with functional assays can help prioritize pharmacogenomic candidate genes . MATERIALS & METHODS : Dose-response experiments were used to quantify the phenotype of sensitivity to DB00544 following the specific knockdown of genes selected from the DB00544 PharmGKB drug pathway in three human colorectal cell lines . Changes in sensitivity were considered significant if the IC(50) for shRNA-exposed cells were three standard deviations outside the mean IC(50) for control-treated cells . RESULTS : Of the 24 genes analyzed , 13 produced significant changes on the phenotype of sensitivity to DB00544 ( P00374 , Q14117 , P23919 , P33316 , Q05932 , Q92820 , P15531 , Q8TCD5 , P23921 , P04818 , Q9BZX2 , P13051 and P11172 ) . CONCLUSION : The RNAi screening strategy enabled prioritization of the genes from the DB00544 drug pathway . Further validation of the genes credentialed in this study should include gene activity or expression and mutation analyses of clinical samples . DB00502 induces neurotoxicity by the DB01221 receptor downstream signaling pathway , alternative from glutamate excitotoxicity . The DB01221 receptor is believed to be important in a wide range of nervous system functions including neuronal migration , synapse formation , learning and memory . In addition , it is involved in excitotoxic neuronal cell death that occurs in a variety of acute and chronic neurological disorders . Besides of agonist/coagonist sites , other modulator sites , including butyrophenone site may regulate the N-methyl-D-aspartate receptor . It has been shown that haloperidol , an antipsychotic neuroleptic drug , interacts with the Q13224 subunit of DB01221 receptor and inhibits DB01221 response in neuronal cells . We found that DB01221 receptor was co-immunoprecipitated by anti-Ras antibody and this complex , beside NR2 subunit of DB01221 receptor contained haloperidol-binding proteins , P29475 and Ras- P01286 . Furthermore , we have shown that haloperidol induces neurotoxicity of neuronal cells via DB01221 receptor complex , accompanied by dissociation of Ras- P01286 from membranes and activation of c-Jun-kinase . Inclusion of insulin prevented relocalization of Ras- P01286 and subsequent neuronal death . DB00502 -induced dissociation of Ras- P01286 leads to inhibition of membrane-bound form of Ras protein and changes downstream regulators activity that results in the initiation of the apoptotic processes via the mitochondrial way . Our results suggest that haloperidol induces neuronal cell death by the interaction with DB01221 receptor , but through the alternative from glutamate excitotoxicity signaling pathway . [ Drugs stimulating insulin release. Importance of their use for improving glycemia , safety and quality of life in diabetes mellitus type 2 ] . Etiopathogenesis of diabetes mellitus is bipolar . On one hand there occurs impairment in beta-cell function caused by genetic factors or abnormal development during fetal period . On the other hand defects of peripheral insulin action are also of significant importance . The bipolarity is also expressed by changing relationship between genetic and environmental factors . P01308 release is connected with closing DB00171 -dependent kalium channel , a structure closely connected with sulfonylurea receptors . Several receptors may be distinguished : Q09428 in Langerhans isles and SUR2 in heart ( SUR2A ) and vessel smoot muscles ( SUR2B ) . In the treatment of insulin release disorders sulfonylureas are still of significant importance though repaglinid and phenyloalanine derivates also have some clinical importance . Within sulfonylurea derivates there have been developed some preparations of slow drug release ( DB01067 GITS , Diaprel MR ) . One daily dose of DB01067 GITS and lower tendency to hypoglycaemia favour acceptation of the therapy by the patients what is also important for their quality of life . Quality of life is now regarded as important as obtaining good indices of diabetes control . A Single 60 mg Dose of DB06643 Might Improve Hepatic P01308 Sensitivity in Postmenopausal Nondiabetic Severe Osteoporotic Women . Background. The O14788 / Q9Y6Q6 / O00300 signaling pathway is crucial for the regulation of osteoclast activity and bone resorption being activated in osteoporosis . The pathway has been also suggested to influence glucose metabolism as observed in chronic low inflammation . Aim . To test whether systemic blockage of O14788 by the monoclonal antibody denosumab influences glucose metabolism in osteoporotic women . Study Design . This is a prospective study on the effect of a subcutaneously injected single 60 mg dose of denosumab in 14 postmenopausal severe osteoporotic nondiabetic women evaluated at baseline and 4 and 12 weeks after their first injection by an oral glucose tolerance test . Results . A single 60 mg dose of denosumab efficiently inhibited serum alkaline phosphatase while it did not exert any significant variation in fasting glucose , insulin , or HOMA-IR at both 4 and 12 weeks . No changes could be detected in glucose response to the glucose load , Matsuda Index , or insulinogenic index . Nonetheless , 60 mg denosumab induced a significant reduction in the hepatic insulin resistance index at 4 weeks and in HbA1c levels at 12 weeks . Conclusions . A single 60 mg dose of denosumab might positively affect hepatic insulin sensitivity though it does not induce clinical evident glucose metabolic disruption in nondiabetic patients . DB00338 , a gastric proton pump inhibitor , inhibits melanogenesis by blocking Q04656 trafficking . DB00338 is a proton pump inhibitor used in the treatment of peptic ulcer disease and gastrosophageal reflux disease and acts by irreversibly blocking P20648 , a P-type H+/K+ ATPase in gastric parietal cells . We found that omeprazole and its closely related congeners inhibited melanogenesis at micromolar concentrations in B16 mouse melanoma cells , normal human epidermal melanocytes , and in a reconstructed human skin model . DB00338 topically applied to the skin of UV-irradiated human subjects significantly reduced pigment levels after 3 weeks compared with untreated controls . DB00338 had no significant inhibitory effect on the activities of purified human tyrosinase or on the mRNA levels of tyrosinase , dopachrome tautomerase , Pmel17 , or O75030 mRNA levels . Although melanocytes do not express P20648 , they do express Q04656 , a copper transporting P-type ATPase in the trans-Golgi network that is required for copper acquisition by tyrosinase . Q04656 relocalization from the trans-Golgi network to the plasma membrane in response to elevated copper concentrations in melanocytes was inhibited by omeprazole . DB00338 treatment increased the proportion of EndoH sensitive tyrosinase , indicating that tyrosinase maturation was impaired . In addition , omeprazole reduced tyrosinase protein abundance in the presence of cycloheximide , suggestive of increased degradation . Our findings are consistent with the hypothesis that omeprazole reduces melanogenesis by inhibiting Q04656 and by enhancing degradation of tyrosinase . P01308 growth factor-receptor ( IGF-1R ) antibody cixutumumab combined with the P42345 inhibitor temsirolimus in patients with refractory Ewing 's sarcoma family tumors . PURPOSE : DB06287 was combined with cixutumumab , a fully human IgG1 monoclonal antibody directed at the insulin growth factor-1 receptor ( IGF-1R ) . EXPERIMENTAL DESIGN : Patients received cixutumumab , 6 mg/kg i.v. weekly , and temsirolimus , 25 to 37.5 mg i.v. weekly ( 4-week cycles ) , with restaging after 8 weeks . Median follow-up was 8.9 months . RESULTS : Twenty patients [ 17 with Ewing 's sarcoma ( Q01844 ) , 3 with desmoplastic small-round cell tumor ( DSRCT ) ] were enrolled . Twelve patients ( 60 % ) were men with a median age of 24 years and six median prior systemic therapies in a metastatic setting . The most frequent toxicities were thrombocytopenia ( 85 % ) , mucositis ( 80 % ) , hypercholesterolemia ( 75 % ) , hypertriglyceridemia ( 70 % ) , and hyperglycemia ( 65 % ; mostly grade I-II ) . Seven of 20 patients ( 35 % ) achieved stable disease ( SD ) for more than 5 months or complete/partial ( CR/PR ) responses . Tumor regression of more than 20 % ( 23 % , 23 % , 27 % , 100 % , 100 % ) occurred in five of 17 ( 29 % ) patients with Q01844 , and they remained on study for 8 to 27 months . One of six patients with Q01844 who previously developed resistance to a different IGF-1R inhibitor antibody achieved a CR . Four of the seven best responders developed grade III mucositis , myelosuppression , or hyperglycemia , which were controlled while maintaining drug dose . CONCLUSION : Cixutumumab combined with temsirolimus was well-tolerated and showed preliminary evidence of durable antitumor activity in heavily pretreated Q01844 family tumors . The P04035 inhibitor pravastatin stimulates insulin secretion through organic anion transporter polypeptides . The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor pravastatin has been reported to have a beneficial effect on reducing the new onset of diabetes as well as lowering plasma lipids . Because pravastatin is a water-soluble organic anion , it can not easily penetrate the lipid bilayer of the cell membrane . As the precise mechanisms of the effect of pravastatin on glucose metabolism and diabetes have not been clarified , we examined the roles of the organic anion transporter family on pravastatin-treated islet and adipocyte functions . Rat oatp1/slco1a1 , oatp2/slco1a4 and oatp3/slco1a5 were expressed in the pancreas , and rat oatp3/slco1a5 was also detected in rat insulinoma cell line P01308 -1e . DB00175 was transported not only by oatp1/slco1a1 and oatp2/slco1a4 , but also by rat oatp3/slco1a5 . DB00175 uptake into P01308 -1e cells was detected and this transport was inhibited by sulfobromophthalein and rifampicin , both of which are known to inhibit oatp family-mediated uptake . In addition , pravastatin enhanced the glucose-stimulated insulin secretion from P01308 -1e cells . When fat-loaded db/db mice were treated with pravastatin , glucose intolerance and insulin resistance were prevented . In addition , insulin secretion from isolated islets was enhanced by pravastatin . These data suggest that pravastatin has pleiotropic effects on islets through membrane transport under high fat/glucose conditions . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . P01308 -like growth factor-1 receptor inhibitor , Q99217 -479 , in cetuximab-refractory head and neck squamous cell carcinoma . BACKGROUND : Recurrent head and neck squamous cell carcinoma ( HNSCC ) remains a difficult cancer to treat . Here , we describe a patient with HNSCC who had complete response to methotrexate ( MTX ) after progressing on multiple cytotoxic agents , cetuximab , and Q99217 -479 ( monoclonal antibody against insulin-like growth factor-1 receptor [ IGF-1R ] ) . METHODS : The clinical information was collected by a retrospective medical record review under an Institutional Review Board-approved protocol . From 4 tumors and 2 normal mucosal epithelia , global gene expression , and IGF-1R and dihydrofolate reductase ( P00374 ) protein levels were determined . RESULTS : Effective target inhibition in the tumor was confirmed by the decreased protein levels of total and phospho-IGF-1R after treatment with Q99217 -479 . Decreased level of P00374 and conversion of a gene expression profile associated with cetuximab-resistance to cetuximab-sensitivity were also observed . CONCLUSION : This suggests that the combination of Q99217 -479 and MTX or cetuximab may be a promising therapeutic approach in refractory HNSCC . Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms . A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells , and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase , is described . DB00120 hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity . The phosphorylation site(s) appear to be the same for both kinases . Phosphorylation is accompanied by increased metabolic flux at low , physiologically relevant , substrate concentrations . P01308 and spermine both inhibit the phosphorylation of the enzyme , possibly by increasing dephosphorylation . P17735 is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium . No parallel changes in flux could be detected . Both enzymes are subject to complex regulatory mechanisms , short- and long-term . Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes . Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux . P01308 reverses growth hormone-induced homologous desensitization . Growth hormone ( GH ) is secreted in a pulsatile pattern to promote body growth and metabolism . GH exerts its function by activating several signaling pathways , including O60674 / P35610 and MEK/ P29323 . P27361 /2 activation by GH plays important roles in gene expression , cell proliferation , and growth . We previously reported that in rat H4IIE hepatoma cells after an initial GH exposure , a second GH exposure induces P42229 phosphorylation but not P27361 /2 phosphorylation ( Ji , S. , Frank , S. J. , and Messina , J. L. ( 2002 ) J. Biol. Chem. 277 , 28384-28393 ) . In this study the mechanisms underlying GH-induced homologous desensitization were investigated . A second GH exposure activated the signaling intermediates upstream of MEK/ P29323 , including O60674 , Ras , and P04049 . This correlated with recovery of P10912 levels , but was insufficient for GH-induced phosphorylation of Q02750 /2 and P27361 /2 . P01308 restored the ability of a second GH exposure to induce phosphorylation of Q02750 /2 and P27361 /2 without altering P10912 levels or GH-induced phosphorylation/activation of O60674 and P04049 . GH and insulin synergized in promoting cell proliferation . Further investigation suggested that insulin increased the amount of MEK bound to Q8IVT5 ( kinase suppressor of Ras ) and restored GH-induced tyrosine phosphorylation of Q8IVT5 . Previous GH exposure also induced desensitization of P42224 and P40763 phosphorylation , but this desensitization was not reversed by insulin . Thus , insulin-regulated resensitization of GH signaling may be necessary to reset the complete response to GH after a normal , physiologic pulse of GH . [ Innate resistance to thymidylate synthase inhibition after 5-fluorouracil treatment -- a rationale of combined use of cisplatin and its optimal administration dose ] . We examined the changes of the number of DB00322 MP binding sites of thymidylate thynthase ( TS-BS ) in Yoshida sarcoma after administration of DB00544 to the tumor bearing rats . We also investigated the optimal dose of DB00515 for the increase of intracellular folate level . In the group received consecutive 7-days administration of DB09327 ( U-7 group ) , total TS-BS was significantly increased compared with non-treatment group and the group received only DB09327 ( U-1 group ) . For free TS-BS , however , there was no difference despite of DB09327 administration . P04818 inhibition rate ( TSIR ) was , therefore , significantly high in U-7 group compared with U-1 group . It seemed necessary to take some counter measure for the induction of TS in the tumor tissue when DB00544 chemotherapy was performed . The optimal dose of DB00515 as a modulator of DB00544 was 1 mg/kg in rat when it was estimated from the changes of intracellular folate levels after administration , which was less than the dose to reveal its own anticancer effect . A new role for the P40763 inhibitor , Q9Y6X2 : a repressor of microphthalmia transcription factor . In vitro and in vivo evidence suggest that microphthalmia transcription factor ( O75030 ) plays a key regulatory role in tissue-specific gene regulation in several cell types , including melanocytes , osteoclasts , and mast cells . A yeast two-hybrid search , using a portion of a nonmutated O75030 gene as the bait in the screening of a mast cell library , resulted in the isolation of the P40763 inhibitor , Q9Y6X2 . Q9Y6X2 is a transcriptional inhibitor that acts by specifically inhibiting P40763 's DNA binding activity . We found that it can directly associate with O75030 using an in vitro pull-down assay . Immunoprecipitation of O75030 from rat basophilic leukemic cells or mouse melanocytes resulted in the specific co-immunoprecipitation of Q9Y6X2 . Co-transfection of O75030 with Q9Y6X2 in NIH 3T3 fibroblasts containing an mMCP-6 promoter-luciferase reporter demonstrated up to 94 % inhibition of O75030 -mediated transcriptional activation . Using a gel-shift assay , it was shown that Q9Y6X2 can block DNA binding activity . It was also found that P40763 does not interfere , either in vitro or in vivo , with the interaction between Q9Y6X2 and O75030 . These data suggest that Q9Y6X2 functions in vivo as a key molecule in supressing the transcriptional activity of O75030 , a role of considerable importance in mast cell and melanocyte development . DB01095 enhances the inhibitory effects of a selective AT1 receptor blocker , valsartan , on atherosclerosis . We investigated the effects of a P04035 inhibitor ( statin ) on the inhibitory effects of an angiotensin II type-1 receptor ( AT1 ) blocker on atherosclerosis and explored cellular mechanisms . We gave apolipoprotein E null mice a high-cholesterol diet for 10 weeks and measured atherosclerotic plaque area and lipid deposition . Neither 1 mg/kg per day of valsartan nor 3 mg/kg per day of fluvastatin had any effect on blood pressure or cholesterol concentration ; however , both drugs decreased plaque area and lipid deposition after 10 weeks . We then reduced the doses of both drugs to 0.1 mg/kg per day and 1 mg/kg per day , respectively . At these doses , neither drug had an effect on atherosclerotic lesions . When both drugs were combined at these doses , a significant reduction in atherosclerotic lesions was observed . Similar inhibitory effects of valsartan or fluvastatin on the expressions of nicotinamide-adenine dinucleotide/nicotinamide-adenine dinucleotide phosphate oxidase subunits P13498 and p47phox , production of superoxide anion , the expression of monocyte chemoattractant protein-1 , and intercellular adhesion molecule-1 expression were observed . These results suggest that concomitant AT1 receptor and cholesterol biosynthesis blockade , particularly when given concomitantly , blunts oxidative stress and inflammation independent of blood pressure or cholesterol-related effects . P01308 like growth factor-1 selectively regulates the expression of matrix metalloproteinase-2 in malignant H-ras transformed cells . The present study demonstrates alterations in the regulation of matrix metalloproteinase-2 ( P08253 ) expression in response to insulin like growth factor-1 ( DB01277 ) in a H-ras transformed cell line , P01024 , which is capable of metastasis formation . These changes in P08253 expression in response to DB01277 treatment did not occur in either non-transformed parental 10 T 1/2 cells or in H-ras transformed cells ( Q13224 cells ) which are capable of benign tumour formation . DB01277 treatment of P01024 cells resulted in increased expression of P08253 gelatinolytic activity and increased expression of P08253 mRNA levels . The DB01277 mediated alterations in P08253 mRNA levels were dependent upon de novo protein synthesis and independent of transcriptional events , but dependent upon post-transcriptional regulatory events . Most notably , DB01277 can regulate P08253 mRNA expression in P01024 cells through a mechanism involving P08253 message stabilization . This study demonstrates aspects of the temporal regulatory mechanisms of P08253 expression in response to insulin-like growth factor-1 in a H-ras transformed fibrosarcoma cell line capable of metastasis formation and thereby , provides further insight into the altered growth regulatory program associated with H-ras mediated cellular transformation and malignant progression .

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[ The importance of biologicals in the treatment of SoJIA ] . Systemic onset juvenile idiopathic arthritis ( SoJIA ) remains difficult to treat . In addition to conventional antirheumatic therapy with non-steroidal antirheumatic drugs ( NSARDs ) , steroids or disease-modifying antirheumatic drugs ( DMARDs ) , biologicals offer a new therapeutic approach for this disease in that they are able to target pathogenically relevant cytokines and effector cells . Some biologicals are already approved for use in children with rheumatic disease.In order to assess the currently available data on the use of biologicals in SoJIA , we performed a Medline search for the period 2005 to March 2010 , including the MeSH terms " SoJIA " , " systemic juvenile idiopathic arthritis " and " biologicals " , as well as an NIH study registry search . At Present there are scant and unconvincing data on the use of DB00005 or DB00051 for the treatment of SoJIA . No results are published on the use of DB00065 or other new P01375 inhibitors . The inhibition of IL-1 or P05231 shows promising results . Data on the efficacy of DB01281 is limited due to very low numbers of SoJIA patients in the studies.Further studies on the use of biologicals in SoJIA while taking individual factors into consideration are required . The long-term safety of all biologicals should be investigated in prospective registers . DB00051 -induced noncaseating granuloma in the bone marrow of a patient being treated for rheumatoid arthritis . Sarcoidosis is a multisystemic disease characterized by noncaseating granulomatous infiltration , primarily of the lungs and lymphatic system . While reports of the efficacy of adalimumab in the treatment of refractory sarcoidosis have been mixed , the more widely used infliximab has demonstrated clear efficacy in this disease . The association between tumor necrosis factor ( P01375 ) -inhibitors and noncaseating granulomas in the lung has been reported in literature . With the exception of one patient treated with adalimumab , who developed pulmonary granuloma , the remaining patients described in literature were treated with etanercept . The current case study is , to our knowledge , the first to describe adalimumab-induced noncaseating granulomas in the bone marrow of a patient being treated for rheumatoid arthritis and suggests that although P01375 -inhibitors are used in the treatment of granulomatous disorders , their use should be carefully monitored as , in rare cases , P01375 -inhibitors may leave sufficient cytokine activation to support granuloma formation . The therapeutic potential of P01375 antagonists for skin psoriasis comorbidities . IMPORTANCE OF THE FIELD : Alopecia , psoriatic arthritis , the metabolic syndrome , inflammatory bowel diseases and cardiovascular diseases may occur as skin psoriatic comorbidities . P01375 antagonists are used to treat psoriasis . DB00051 is one of the recognized active agents for this indication . AREAS COVERED IN THIS REVIEW : The current peer-reviewed publications and presentation of original findings . WHAT THE READER WILL GAIN : DB00051 is active on recalcitrant psoriasis and some of its comorbidities , particularly arthropathies and Crohn 's disease . However , the progression of the radiological alterations is limited with regression of the bony erosions . Psoriatic enthesopathy also regresses . Mortality associated with psoriasis arthropathy is on the decline . Crohn 's disease , the most frequent inflammatory bowel comorbidity of psoriasis , is responsive to adalimumab . The effect of adalimumab on the metabolic syndrome and cardiovascular involvement is more erratic . The spectacular effects of adalimumab may be associated with some adverse effects . In particular , despite a marked reduction in the psoriasis area-and-severity index ( PASI ) score some new acute lesions of cutaneous psoriasis may develop corresponding to paradoxical psoriasis . Other potential adverse effects include infections , granulomas , rapid growth of cancers and occurrence of lymphomas . TAKE HOME MESSAGE : DB00051 frequently controls moderate-to-severe forms of cutaneous psoriasis and some of its comorbidities . [ Cutaneous infection due to Mycobacterium chelonae during anti- P01375 therapy ] . BACKGROUND : Mycobacterium chelonae is a ubiquitous , rapidly growing , opportunistic , non-tuberculous mycobacterium that can cause skin and bone tissue infections . We report a case of cutaneous infection due to M. chelonae following anti- P01375 therapy . CASE REPORT : A 70-year-old woman with a medical history of rheumatoid arthritis was admitted for several purple nodular cutaneous lesions on her right leg evolving for 2 months . At admission , she was on prednisone , methotrexate and adalimumab for her rheumatoid arthritis . Skin lesions appeared 5 days before etanercept , which was taken for 5 months before being discontinued for adalimumab . Both the histopathological examination and bacterial culture of involved skin showed the presence of M. chelonae . DB00051 was immediately discontinued and a combination of amoxicillin-clavulanic acid and tigecyclin was started . DISCUSSION : P01375 plays a pivotal role in immune reaction to intracellular pathogens . Very few cases of cutaneous infection involving M. chelonae in association with an anti- P01375 therapy have been reported in the literature . To our knowledge , this is the first case occurring during treatment with etanercept and symptoms worsened with the introduction of adalimumab . In addition , this case underlines the difficulties of effectively treating this mycobacterium . DB00051 for the treatment of rheumatoid arthritis . Rheumatoid arthritis ( RA ) is characterized by massive synovial proliferation resulting in joint destruction and deterioration in quality of life . P01375 has been implicated in the central pathogenesis of RA , and its blockade ? by anti- P01375 monoclonal antibodies ( infliximab and adalimumab ) and the soluble P01375 receptor ( etanercept ) has caused a paradigm shift in the treatment of RA . DB00051 can be used alone or concomitantly with methotrexate , but the combination of both is significantly superior to monotherapy with adalimumab . DB00051 can not only ameliorate the signs and symptoms of the disease , but can also inhibit the progression of joint destruction and improve quality of life . Approximately 50 % of patients with early , aggressive RA receiving combination therapy with adalimumab and methotrexate can be introduced into disease remission ( disease activity score < 2.6 ) . DB00051 is even effective in patients with RA treated previously with other biologics . Long-term treatment with adalimumab is safe and well tolerated in patients with RA . [ DB00051 ] . Recently , the anti-tumor necrosis factor( P01375 )-alpha treatments for RA are successful in alleviating the discomforts associated with swollen , painful joints . DB00051 (D2E7) is the first fully human anti- P01375 monoclonal antibody(IgG1) . Therefore , it has low immunogenicity and possibly greater therapeutic potential compared with other anti- P01375 antibodies . This is administered subcutaneously at a dose of 1 mg/kg biweekly . The combined therapy with methotrexate(MTX) is efficacious to the patients who receive MTX alone and are insufficient to control symptoms of RA . The therapeutic effects become evident within 24 hours to one week after administration and reached maximum effect after one to two weeks . In adalimumab recipient , radiographic progression is also controlled and serum levels of matrix metalloproteinase-1( P03956 ) and P08254 decrease . For patients with RA , the treatment of adalimumab will set a new standard for symptom control and joint protection . Human anti-tumor necrosis factor monoclonal antibody ( adalimumab ) in Crohn 's disease : the CLASSIC-I trial . BACKGROUND & AIMS : P01375 blockade has been shown to be an effective treatment strategy in Crohn 's disease ( CD ) . DB00051 is a human immunoglobulin P55008 ( IgG(1) ) monoclonal antibody targeting tumor necrosis factor ( P01375 ) . A randomized , double-blind , placebo-controlled , dose-ranging trial was performed to evaluate the efficacy of adalimumab induction therapy in patients with CD . METHODS : A total of 299 patients with moderate to severe CD naive to anti- P01375 therapy were randomized to receive subcutaneous injections at weeks 0 and 2 with adalimumab 40 mg/20 mg , 80 mg/40 mg , or 160 mg/80 mg or placebo . The primary endpoint was demonstration of a significant difference in the rates of remission at week 4 ( defined as a Crohn 's Disease Activity Index score < 150 points ) among the 80 mg/40 mg , 160 mg/80 mg , and placebo groups . RESULTS : The rates of remission at week 4 in the adalimumab 40 mg/20 mg , 80 mg/40 mg , and 160 mg/80 mg groups were 18 % ( P = .36 ) , 24 % ( P = .06 ) , and 36 % ( P = .001 ) , respectively , and 12 % in the placebo group . Adverse events occurred at similar frequencies in all 4 treatment groups except injection site reactions , which were more common in adalimumab-treated patients . CONCLUSIONS : DB00051 was superior to placebo for induction of remission in patients with moderate to severe Crohn 's disease naive to anti- P01375 therapy . The optimal induction dosing regimen for adalimumab in this study was 160 mg at week 0 followed by 80 mg at week 2 . DB00051 was well tolerated . [ P35354 inhibitor non-steroidal anti-inflammatory drugs , myth or reality ? ] . The discovery of two isoforms of cyclooxygenase , Cox-1 constitutive and Cox-2 inducible , has prompted the development of new molecules with high Cox-2 selectivity . These new NSAIDs belong to the coxib class and have theoretically a better digestive tolerability than classical NSAID have . In Belgium , rofecoxib ( ( Vioxx ) and celecoxib ( DB00482 ) are commercialized . DB00533 is indicated in the symptomatic treatment of osteoarthritis ( 12.5 to 25 mg/d ) and celecoxib is indicated in osteoarthritis ( 200 mg/d ) and in rheumatoid arthritis ( 200 to 400 mg/d ) . Several studies have demonstrated their efficacy , similarly to classical NSAID as diclofenac ( Voltaren ) , naproxen ( Naprosyne ) , ibuprofen ( DB01050 ) and their superiority compared to placebo . Their safety profile for gastrointestinal events is proven in patients without ulcer history compared to classical NSAID . However , the concomitant use of aspirin decreases the benefit as demonstrated for celecoxib at 400 mg/d but not investigated for rofecoxib . The selective inhibition of Cox-2 with no effect on Cox-1 favors cardiovascular events in patients at risk . Other side effects are similar to classical NSAID . Thus Cox-2 inhibitors NSAID are interesting molecules for their sparing gastrointestinal activity . They must be used with caution in patients with ulcer history , in the elderly and in patients requiring aspirin for cardiovascular prophylaxis . Selective targeting of the repressive transcription factors P25490 and cMyc to disrupt quiescent human immunodeficiency viruses . Quiescent HIV-1 infection of resting P01730 (+) T cells is an obstacle to eradication of HIV-1 infection . These reservoirs are maintained , in part , by repressive complexes that bind to the HIV-1 long terminal repeat ( LTR ) and recruit histone deacetylases (HDACs). cMyc and P25490 are two transcription factors that are recruited as part of well-described , distinct complexes to the HIV-1 LTR and in turn recruit HDACs . In prior studies , depletion of single factors that recruit Q13547 in various cell lines was sufficient to upregulate LTR activity . We used short hairpin RNAs ( shRNAs ) to test the effect of targeted disruption of a single transcription factor on quiescent proviruses in T cell lines . In this study , we found that depletion of P25490 significantly increases mRNA and protein expression from the HIV-1 promoter in some contexts , but does not affect Q13547 , Q92769 , O15379 , or acetylated histone 3 occupancy of the HIV-1 LTR . Conversely , depletion of cMyc or cMyc and P25490 does not significantly alter the level of transcription from the LTR or affect recruitment of HDACs to the HIV-1 LTR . Furthermore , global inhibition of HDACs with the HDAC inhibitor suberoylanilide hydroxamic acid ( DB02546 ) enhanced the increase in LTR transcription in cells that were depleted of P25490 .These findings show that despite prior isolated findings , redundancy in repressors of HIV-1 LTR expression will require selective targeting of multiple restrictive mechanisms to comprehensively induce the escape of quiescent proviruses from latency . [ Genetic polymorphisms and cancer risk ] . While hereditary disease genes have a high lifelong cumulative incidence rate ( penetrance ) , the penetrance for polymorphism genotypes is not high . Polymorphisms relating to cancer incidence are classified into 1. carcinogen metabolizing enzymes ( CYPs , GSTs , P15559 , etc. ) , 2 . DNA repair enzymes ( O15527 , P18887 , P18074 , etc. ) , 3 . DNA synthesis and methylation ( P42898 , MS , etc. ) , 4. cytokines and inflammation-related enzymes ( IL-1B , P01375 -A , P05164 , etc. ) , and 5. sex hormone metabolizing enzymes and the receptors ( P11511 , P31213 , ER , etc. ) . Since genotypes can not be manipulated , they are not the factors subject to prevention . However , the finding that the strength of association between lifestyle and disease occurrence is influenced by genotypes ( gene-environment interaction ) , opens the door to genotype applications for disease prevention practice . DB00051 in juvenile rheumatoid arthritis/juvenile idiopathic arthritis . Chronic arthritis in childhood is the most common pediatric rheumatic disease and can lead to significant short- and long-term disability . P01375 is a cytokine involved in joint inflammation and destruction . It has been suggested that early and aggressive treatment leads to improved outcomes by ameliorating clinical signs and symptoms , inhibiting joint destruction and improving functional disability . The early and aggressive use of disease-modifying antirheumatic drugs and biologic response modifiers are key to achieving this goal . Anti- P01375 agents are effective biologic modifiers that have had a major impact on the clinical course of arthritis . DB00051 , a recombinant monoclonal antibody to P01375 , is emerging as a valuable new therapy for juvenile arthritis . DB00051 -induced acute interstitial lung disease in a patient with rheumatoid arthritis . The use of immunobiological agents for the treatment of autoimmune diseases is increasing in medical practice . Anti- P01375 therapies have been increasingly used in refractory autoimmune diseases , especially rheumatoid arthritis , with promising results . However , the use of such therapies has been associated with an increased risk of developing other autoimmune diseases . In addition , the use of anti- P01375 agents can cause pulmonary complications , such as reactivation of mycobacterial and fungal infections , as well as sarcoidosis and other interstitial lung diseases ( ILDs ) . There is evidence of an association between ILD and the use of anti- P01375 agents , etanercept and infliximab in particular . DB00051 is the newest drug in this class , and some authors have suggested that its use might induce or exacerbate preexisting ILDs . In this study , we report the first case of acute ILD secondary to the use of adalimumab in Brazil , in a patient with rheumatoid arthritis and without a history of ILD . Effective long-term control of refractory hidradenitis suppurativa with adalimumab after failure of conventional therapy . BACKGROUND : Hidradenitis suppurativa is a chronic suppurative condition featuring inflammatory nodules , fistulas and scars . It occurs predominantly in the axillae and groin . The disease is poorly responsive to any treatment and is connected with significant morbidity . Systemic therapy , including oral antibiotics , retinoids and antiandrogens , usually has only limited effect . Surgical treatment of affected areas is necessary in advanced stages . OBJECTIVES : Several reports support the beneficial effect of tumor necrosis factor-α ( P01375 -α ) antagonists for the treatment of severe hidradenitis suppurativa . By contrast with data on infliximab and etanercept , data describing the potential positive influence of adalimumab on disease outcome are limited and refer to only small cohorts of patients . METHODS : Eight patients with severe , recalcitrant hidradenitis were treated for 1 year with adalimumab in a standard regimen and were subsequently followed for 1 year . RESULTS : All patients improved within 4-6 weeks and laboratory parameters of P02741 ( CRP ) and leukocyte count reduced significantly during treatment . Three patients demonstrated long-lasting improvement and five showed recurrences several months after discontinuation of the therapy . The average recurrence-free interval was 9.5 months . CONCLUSIONS : DB00051 is suitable for the long-term treatment of hidradenitis suppurativa and presents a further conservative treatment approach . [ Biologics in the early treatment of ankylosing spondylitis and related forms of spondyloarthritis ] . New pathogenetic insights have identified the key role of P01375 in inflammatory rheumatic diseases and have revolutionized the therapy of spondyloarthritides . P01375 -antagonists specifically inhibit the pro-inflammatory effects of P01375 . Clinical studies with infliximab ( Remicade ) , DB00005 ( Enbrel ) or DB00051 ( Humira ) in ankylosing spondylitis or related diseases demonstrate superior efficacy to conventional drugs like non-steroidal antirheumatic drugs or traditional disease modifying antirheumatic drugs . Targeting Q01196 / Q06455 -histone deacetylase repressor complex : a novel mechanism for valproic acid-mediated gene expression and cellular differentiation in Q01196 / Q06455 -positive acute myeloid leukemia cells . In t(8;21) acute myeloid leukemia ( AML ) , the Q01196 / Q06455 fusion protein promotes leukemogenesis by recruiting class I histone deacetylase ( HDAC ) -containing repressor complex to the promoter of Q01196 target genes . Valproic acid ( DB00313 ) , a commonly used antiseizure and mood stabilizer drug , has been shown to cause growth arrest and induce differentiation of malignant cells via HDAC inhibition . DB00313 causes selective proteasomal degradation of Q92769 but not other class I HDACs ( i.e. , HDAC 1 , 3 , and 8 ) . Therefore , we raised the question of whether this drug can effectively target the leukemogenic activity of the Q01196 / Q06455 fusion protein that also recruits Q13547 , a key regulator of normal and aberrant histone acetylation . We report here that DB00313 treatment disrupts the Q01196 / Q06455 - Q13547 physical interaction , stimulates the global dissociation of Q01196 / Q06455 - Q13547 complex from the promoter of Q01196 / Q06455 target genes , and induces relocation of both Q01196 / Q06455 and Q13547 protein from nuclear to perinuclear region . Furthermore , we show that mechanistically these effects associate with a significant inhibition of HDAC activity , histone H3 and H4 hyperacetylation , and recruitment of RNA polymerase II , leading to transcriptional reactivation of target genes ( i.e. , P08700 ) otherwise silenced by Q01196 / Q06455 fusion protein . Ultimately , these pharmacological effects resulted in significant antileukemic activity mediated by partial cell differentiation and caspase-dependent apoptosis . Taken together , these data support the notion that DB00313 might effectively target Q01196 / Q06455 -driven leukemogenesis through disruption of aberrant Q13547 function and that DB00313 should be integrated in novel therapeutic approaches for Q01196 / Q06455 -positive AML . DB00051 ( antitumour necrosis factor-α ) treatment of hidradenitis suppurativa ameliorates skin inflammation : an in situ and ex vivo study . BACKGROUND : Hidradenitis suppurativa ( HS ) is a difficult-to-manage disease . Randomized controlled trials with antitumour necrosis factor ( P01375 ) -α biologics have been conducted and in most studies disease activity was reduced . However , the mechanism of action in HS skin is so far unknown . OBJECTIVES : To assess whether anti- P01375 -α treatment affects in situ cytokine production and frequency of inflammatory cell populations in HS lesional skin . METHODS : Nine patients with HS , participating in a larger placebo-controlled , double-blind phase IIb clinical trial on the efficacy and safety of adalimumab in patients with moderate to severe HS ( M10-467 ) , were randomized and treated for 16weeks . In a mechanism-of-action substudy , biopsies were obtained at fixed time points pre- and post-treatment . One part of the biopsy was cultured for 24h for cytokine release in the culture medium , while another part was used for in situ analysis . RESULTS : Secretion of cytokines , including interleukin ( IL ) -1β , Q07325 [ monokine induced by interferon-γ ( Q07325 ) ] , P22301 , IL-11 , B-lymphocyte chemoattractant ( O43927 ) and Q16552 , was significantly elevated in HS . DB00051 treatment was associated with decreased production of cytokines in HS skin , especially IL-1β , Q07325 ( Q07325 ) and O43927 . Treatment significantly reduced the number of CD11c+, P08571 + and P34810 + cells in HS lesional skin . The numbers of CD3+ and P01730 + T cells , and P11836 + and CD138+ B cells were also reduced by adalimumab treatment . CONCLUSIONS : DB00051 treatment inhibits important cytokines and inflammatory cell numbers in lesional HS skin , especially levels of IL-1β and numbers of inflammatory CD11c+ dendritic cells . [ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC/MS/MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r=0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC/MS/MS analysis ( r=0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations . DB00051 ( P01375 α Inhibitor ) Therapy Exacerbates IgA Glomerulonephritis Acute Renal Injury and Induces Lupus Autoantibodies in a Psoriasis Patient . DB00051 ( Humira ) is a tumour necrosis factor α ( P01375 α ) inhibitor that is approved for the treatment of rheumatoid arthritis , psoriasis , psoriatic arthritis , Crohn 's disease , ankylosing spondylitis , and juvenile idiopathic arthritis ( Sullivan and Preda ( 2009 ) , Klinkhoff ( 2004 ) , and Medicare Australia ) . Use of P01375 α inhibitors is associated with the induction of autoimmunity ( systemic lupus erythematosus , vasculitis , and sarcoidosis or sarcoid-like granulomas ) ( Ramos-Casals et al. ( 2010 ) ) . We report a patient with extensive psoriasis presenting with renal failure and seropositive lupus markers without classical lupus nephritis after 18 months treatment with adalimumab . He has renal biopsy proven IgA nephritis instead . Renal biopsy is the key diagnostic tool in patients presenting with adalimumab induced nephritis and renal failure . He made a remarkable recovery after adalimumab cessation and steroid treatment . To our knowledge , this is a unique case of a psoriasis patient presenting with seropositive lupus markers without classical lupus nephritis renal failure and had renal biopsy proven IgA glomerulonephritis after receiving adalimumab . DB01296 sulfate inhibits P01375 and P01579 -induced production of P05362 in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) -1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE-19 cells were used as a model to determine the effects of GS on the expression of the P05362 gene upregulated by P01375 or P01579 , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 and P01579 increased the expression of P05362 at the mRNA and protein levels in a time- and dose-dependent manner in ARPE-19 cells . GS effectively downregulated the P01375 - or P01579 -induced expression of P05362 in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 and phosphorylated P42224 in P01579 -stimulated ARPE-19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 gene in ARPE-19 cell stimulated with P01375 or P01579 through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 . This study has demonstrated a potentially important property of GS in reducing P05362 mediated inflammatory mechanisms in the eye . The p65 ( RelA ) subunit of NF-kappaB interacts with the histone deacetylase ( HDAC ) corepressors Q13547 and Q92769 to negatively regulate gene expression . Regulation of NF-kappaB transactivation function is controlled at several levels , including interactions with coactivator proteins . Here we show that the transactivation function of NF-kappaB is also regulated through interaction of the p65 ( RelA ) subunit with histone deacetylase ( HDAC ) corepressor proteins . Our results show that inhibition of HDAC activity with trichostatin A ( P32119 ) results in an increase in both basal and induced expression of an integrated NF-kappaB-dependent reporter gene . Chromatin immunoprecipitation ( ChIP ) assays show that P32119 treatment causes hyperacetylation of the wild-type integrated NF-kappaB-dependent reporter but not of a mutant version in which the NF-kappaB binding sites were mutated . Expression of Q13547 and Q92769 repressed tumor necrosis factor ( P01375 ) -induced NF-kappaB-dependent gene expression . Consistent with this , we show that Q13547 and Q92769 target NF-kappaB through a direct association of Q13547 with the Rel homology domain of p65 . Q92769 does not interact with NF-kappaB directly but can regulate NF-kappaB activity through its association with Q13547 . Finally , we show that inhibition of HDAC activity with P32119 causes an increase in both basal and P01375 -induced expression of the NF-kappaB-regulated interleukin-8 ( P10145 ) gene . Similar to the wild-type integrated NF-kappaB-dependent reporter , ChIP assays showed that P32119 treatment resulted in hyperacetylation of the P10145 promoter . These data indicate that the transactivation function of NF-kappaB is regulated in part through its association with HDAC corepressor proteins . Moreover , it suggests that the association of NF-kappaB with the Q13547 and Q92769 corepressor proteins functions to repress expression of NF-kappaB-regulated genes as well as to control the induced level of expression of these genes . DB00452 -arginine conjugate , a novel HIV-1 Tat antagonist : synthesis and anti-HIV activities . HIV-1 transactivating protein Tat is essential for virus replication and progression of HIV disease . HIV-1 Tat stimulates transactivation by binding to HIV-1 transactivator responsive element ( TAR ) RNA , and while secreted extracellularly , it acts as an immunosuppressor , an activator of quiescent T-cells for productive HIV-1 infection , and by binding to CXC chemokine receptor type 4 ( P61073 ) as a chemokine analogue . Here we present a novel HIV-1 Tat antagonist , a neomycin B-hexaarginine conjugate ( NeoR ) , which inhibits Tat transactivation and antagonizes Tat extracellular activities , such as increased viral production , induction of P61073 expression , suppression of CD3-activated proliferation of lymphocytes , and upregulation of the CD8 receptor . Moreover , Tat inhibits binding of fluoresceine isothiocyanate ( FITC ) -labeled NeoR to human peripheral blood mononuclear cells ( PBMC ) , indicating that Tat and NeoR bind to the same cellular target . This is further substantiated by the finding that NeoR competes with the binding of monoclonal Abs to P61073 . Furthermore , NeoR suppresses HIV-1 binding to cells . Importantly , NeoR accumulates in the cell nuclei and inhibits the replication of M- and T-tropic HIV-1 laboratory isolates ( EC(50) = 0.8-5.3 microM ) . A putative model structure for the TAR-NeoR complex , which complies with available experimental data , is presented . We conclude that NeoR is a multitarget HIV-1 inhibitor ; the structure , and molecular modeling and dynamics , suggest its binding to TAR RNA . NeoR inhibits HIV-1 binding to cells , partially by blocking the P61073 HIV-1 coreceptor , and it antagonizes Tat functions . NeoR is therefore an attractive lead compound , capable of interfering with different stages of HIV infection and AIDS pathogenesis . DB00051 in the management of Behçet 's disease . Behçet 's disease ( BD ) is a relapsing , systemic , inflammatory disorder that affects various organ systems . Most of the manifestations of BD are self-limiting , but ocular attacks are an exception . Gastrointestinal tract , central nervous system , and cardiovascular system manifestations are relatively infrequent but may be resistant to conventional immunosuppressive treatment and therefore life-threatening . P01375 alpha antagonists are increasingly being used in patients whose BD is inadequately controlled by standard immunosuppressive regimens . Most of the current experience regarding the treatment of refractory BD involves the use of infliximab ; however , adalimumab has also been successfully used in cases of BD refractory to both conventional therapy and infliximab . Compared with infliximab , adalimumab offers several other advantages , such as the ability to self-administer at home , better patient compliance , and an improved side effect profile . Here , we review clinical experience of the use of adalimumab to treat the serious manifestations of BD . DB00051 is a promising drug for the treatment of BD , and its randomized , prospective study in a large number of patients is warranted to fully determine its efficacy in the refractory BD setting . Novel marine phenazines as potential cancer chemopreventive and anti-inflammatory agents . Two new ( 1 and 2 ) and one known phenazine derivative ( lavanducyanin , 3 ) were isolated and identified from the fermentation broth of a marine-derived Streptomyces sp . ( strain CNS284 ) . In mammalian cell culture studies , compounds 1 , 2 and 3 inhibited P01375 -α-induced NFκB activity ( IC₅₀ values of 4.1 , 24.2 , and 16.3 μM , respectively ) and LPS-induced nitric oxide production ( IC₅₀ values of > 48.6 , 15.1 , and 8.0 μM , respectively ) . PGE₂ production was blocked with greater efficacy ( IC₅₀ values of 7.5 , 0.89 , and 0.63 μM , respectively ) , possibly due to inhibition of cyclooxygenases in addition to the expression of P35354 . Treatment of cultured HL-60 cells led to dose-dependent accumulation in the subG1 compartment of the cell cycle , as a result of apoptosis . These data provide greater insight on the biological potential of phenazine derivatives , and some guidance on how various substituents may alter potential anti-inflammatory and anti-cancer effects . DB00051 : new indication . Ankylosing spondylitis : just another P01375 alpha antagonist . No comparisons with other P01375 alpha antagonists . Mucosal expression of basic fibroblastic growth factor , syndecan 1 and tumour necrosis factor-α in Crohn 's disease in deep remission under treatment with anti-TNFα antibodies . BACKGROUND AND AIMS : Both inflammation and fibrosis may be detected in Crohn 's disease ( CD ) . The molecular pattern of Basic Fibroblastic Growth Factor ( P09038 ) and P18827 ( SD1 ) expression is altered in stenosing CD , but we do not know what the behaviour of this teamwork factor is in CD in deep remission under treatment with anti-TNFα antibodies . Our aim was to compare the expression of P09038 , SD1 and P01375 -α in patients with CD in deep remission under treatment with DB00065 ( IFX ) or DB00051 ( P00813 ) and a control group of patients with active CD . METHODS : We assessed the expression of P09038 , SD1 and P01375 -α in 10 patients with active CD and in 28 patients with CD in sustained deep remission for at least 6 months . All patients underwent surveillance colonoscopy with biopsies , while receiving maintenance therapy with IFX or P00813 . Analysis was conducted by real-time reverse transcriptase PCR ( RT-PCR ) in biopsy samples . RESULTS : We found that P09038 , SD1 and P01375 -α were significantly reduced under treatment with anti-TNFα versus controls ( p=0.000 ) . P09038 and SD1 expression were similar between IFX and P00813 patients ( p=0.335 and p=0.289 , respectively ) , while P01375 -α was significantly under-expressed in P00813 patients ( p=0.008 ) . CONCLUSIONS : P09038 , SD1 and P01375 -α are significantly reduced in CD patients in deep remission under treatment with anti-TNFα , likely as an expression of optimal control of inflammation . The significance of the P01375 -α under-expression in patients under treatment with P00813 with respect to those under treatment with IFX should be elucidated in further studies . Thalidomide suppresses Up-regulation of human immunodeficiency virus coreceptors P61073 and P51681 on P01730 + T cells in humans . Concurrent infection in patients with human immunodeficiency virus ( HIV ) infection increases the expression of HIV coreceptors P61073 and P51681 . Thalidomide has beneficial effects in a number of HIV-associated diseases . The effect of thalidomide on P61073 and P51681 expression on P01730 + T cells was determined . Thalidomide produced a dose-dependent inhibition of lipopolysaccharide ( LPS ) -induced up-regulation of P61073 and P51681 in vitro . Antibody to tumor necrosis factor-alpha ( P01375 ) also attenuated the LPS-induced HIV coreceptor up-regulation , which was not further reduced by thalidomide . Thalidomide ( 400 mg ) was orally administered to 6 men , and their blood was stimulated ex vivo with LPS , staphylococcal or mycobacterial antigens , or antibody to CD3 or P10747 cells . All stimuli induced up-regulation of HIV coreceptors , which was reduced after ingestion of thalidomide . Thalidomide may be beneficial in the treatment of intercurrent infections during HIV infection by reducing the up-regulation of P61073 and P51681 expression on P01730 + T cells induced by bacterial and mycobacterial antigens , by a mechanism that involves inhibition of P01375 . Characterization of plant P18887 and its interaction with proliferating cell nuclear antigen . In plants , there are no P06746 ( Pol beta ) and P49916 ( Lig3 ) genes . Thus , the plant short-patch base excision repair ( short-patch BER ) pathway must differ considerably from that in mammals . We characterized the rice ( Oryza Sativa L. cv. Nipponbare ) homologue of the mammalian X-ray repair cross complementing 1 ( P18887 ) , a well-known BER protein . The plant P18887 lacks the N-terminal domain ( NTD ) which is required for Pol beta binding and is essential for mammalian cell survival . The recombinant rice P18887 ( OsXRCC1 ) protein binds single-stranded DNA ( ssDNA ) as well as double-stranded DNA ( dsDNA ) and also interacts with rice proliferating cell nuclear antigen ( OsPCNA ) in a pull-down assay . Through immunoprecipitation , we demonstrated that OsXRCC1 forms a complex with P12004 in vivo . OsXRCC1 mRNA was expressed in all rice organs and was induced by application of bleomycin , but not of MMS , H(2)O(2) or UV-B . DB00290 also increased the fraction of OsXRCC1 associated with chromatin . These results suggest that OsXRCC1 contributes to DNA repair pathways that differ from the mammalian BER system . DB08879 -- an anti- Q9Y275 human monoclonal antibody for rheumatoid arthritis . INTRODUCTION : Q9Y275 ( Q9Y275 ) is a major regulatory factor that controls the development and survival of B cells . Elevated serum levels of Q9Y275 have been associated with rheumatoid arthritis ( RA ) . DB08879 is a fully human monoclonal antibody that inhibits Q9Y275 and it is being developed for the treatment of RA . This review aims to summarize up-to-date pharmacological and clinical data of belimumab in the treatment of RA . AREAS COVERED : A literature search was performed on PubMed using keywords , including belimumab , LymphoStat-B , benlysta , Q9Y275 inhibitor , rheumatoid arthritis and autoimmune disease . References of relevant studies were searched by hand . Abstracts of international conferences up to October 2012 were also included . DB08879 was well tolerated in the treatment of RA over 24 weeks . It significantly increased American College of Rheumatology ( P10323 )20 responses at week 24 , especially in patients with high disease activity , positive rheumatoid factor , no anti- P01375 treatment experience and those who had failed methotrexate therapy . However , belimumab failed to demonstrate significantly improved ACR50 and ACR70 responses in the single Phase II clinical trial of RA . EXPERT OPINION : These results suggest that the clinical efficacy of belimumab for RA needs to be further investigated in future clinical trials . Careful patient selection may be necessary for belimumab to achieve optimal clinical outcomes in RA . Treatment of ankylosing spondylitis with P01375 blockers : a meta-analysis . Biological agents directed against tumor necrosis factor ( P01375 ) represent therapeutic options for patients with ankylosing spondylitis with high disease activity despite use of non-steroidal anti-inflammatory drugs . To evaluate the efficacy and safety of the anti- P01375 agents infliximab , etanercept , adalimumab , DB06674 , and certolizumab for the treatment of ankylosing spondylitis , we performed a systematic review of randomized clinical trials on adult patients with ankylosing spondylitis using articles culled from the EMBASE , MEDLINE , Cochrane Controlled Trials Register and LILACS databases ( September/2012 ) , manual literature search , and the gray literature . Study selections and data collection were performed by two independent reviewers , with disagreements solved by a third reviewer . The following outcomes were evaluated : ASAS 20 response , disease activity , physical function , vertebral mobility , adverse events , and withdraws . The meta-analysis was performed using the Review Manager(®) 5.1 software by applying the random effects model . Eighteen studies were included in this review . No study of certolizumab was included . Patients treated with anti- P01375 agents were more likely to display an ASAS 20 response after 12/14 weeks ( RR 2.21 ; 95 % CI 1.91 ; 2.56 ) and 24 weeks ( RR 2.68 ; 95 % CI 2.06 ; 3.48 ) compared with controls , which was also true for several other efficacy outcomes . Meta-analysis of safety outcomes and withdraws did not indicate statistically significant differences between treatment and control groups after 12 or 30 weeks . DB00051 , infliximab , etanercept , and DB06674 can effectively reduce the signs and symptoms of the axial component of ankylosing spondylitis . Safety outcomes deserve further study , especially with respect to long-term follow-ups . DB00051 , etanercept , infliximab , and the risk of tuberculosis : data from clinical trials , national registries , and postmarketing surveillance . This review evaluates the risk of tuberculosis ( TB ) , adherence with recommendations for TB prevention , and host-related risk in patients with rheumatoid arthritis ( RA ) , psoriatic arthritis , and ankylosing spondylitis receiving infliximab ( IFX ) , adalimumab ( P00813 ) , and etanercept ( DB01613 ) through an analysis of phase III randomized controlled trials ( RCT ) , postmarketing surveillance , and national registries . Ten ( 0.21 % ) TB cases occurred among 4590 patients in 16 RCT of IFX , 9 ( 0.12 % ) among 7009 patients in 21 RCT of P00813 , and 4 ( 0.05 % ) among 7741 patients in 26 RCT of DB01613 . Overall , 19/23 ( 83 % ) TB cases occurred in patients with RA . Data from national registries and postmarketing surveillance showed an increased risk of TB in patients receiving any of the 3 anti-tumor necrosis factor ( P01375 ) drugs , with a 3-4 times higher risk associated with IFX and P00813 than with DB01613 . Deviations from recommended TB prevention procedures were observed in up to 80 % of patients , and most registries did not include data on host-related risk factors for TB . TB occurrence was reduced in recent RCT but not in real-life practice . TB risk was lower for DB01613 than for monoclonal antibody anti- P01375 agents . More complete data collection , including host-related TB risk factors , is advisable to avoid biased results . DB00051 -associated antiphospholipid syndrome : a case report and review of the literature . This study aims for the presentation of the first reported case of adalimumab-associated antiphospholipid syndrome ( APS ) and review of the literature on adalimumab-induced vasculitis and APS . A case of APS associated with adalimumab use in a 67-year-old woman is reported . The English medical literature was reviewed for antitumor necrosis factor ( P01375 ) agents and their association with APS and vasculitis . DB00051 is a fully humanized monoclonal antibody targeted against P01375 alpha that is widely used in the treatment of rheumatoid arthritis , juvenile idiopathic arthritis , ankylosing spondylitis , psoriatic arthritis , psoriasis , and Crohn 's disease . Literature review reveals several cases of anti- P01375 -induced vasculitis including cases associated with adalimumab . We report the first case of adalimumab-induced APS in a 67-year-old woman who developed APS and vasculitis associated with de novo positive anti-cardiolipin ( aCL ) antibody following the third dose of adalimumab therapy for the treatment of spondyloarthropathy . This is the first case demonstrating that a short course of adalimumab therapy may induce immunoglobulin M aCL autoantibodies leading to APS . With the growing use of anti- P01375 medications in immune-mediated and inflammatory diseases , adalimumab and other anti- P01375 medications should be considered as a possible explanation for APS . DB00051 therapy for refractory uveitis : results of a multicentre , open-label , prospective trial . OBJECTIVE : Tumour necrosis factor ( P01375 ) blockers have been demonstrated to be effective in the treatment of systemic and ocular inflammatory diseases . We conducted a prospective , multicentre , open-label Phase II clinical trial to assess the effectiveness and safety of adalimumab , a fully human anti- P01375 monoclonal antibody , in treating refractory uveitis . METHODS : Subjects with non-infectious uveitis refractory to corticosteroids and at least one other immunosuppressive medication were enrolled . Treatment outcome was ascertained by a composite endpoint comprised of visual acuity , intraocular inflammation , ability to taper immunosuppressives , and posterior segment imaging . Clinical response was defined by improvement in at least one parameter , worsening in none , and well controlled intraocular inflammation . Week 10 responders were permitted to continue receiving adalimumab for the study duration of 50 weeks . RESULTS : Twenty-one of 31 patients ( 68 % ) were characterised as clinical responders at 10 weeks , of whom 12 patients ( 39 % ) exhibited durable response after 50 weeks . The most common reason for study termination was primary or secondary inefficacy . No patients experienced treatment-limiting toxicity clearly related to study therapy . CONCLUSIONS : DB00051 was safe and effective in 68 % of refractory uveitis patients 10 weeks after study enrolment , and maintained in 39 % after 1 year . Ongoing study is required to determine the place of adalimumab and other P01375 blockers in the treatment of uveitis . Probable diffuse retinopathy caused by adalimumab in a patient with Crohn 's disease . DB00051 is a tumor necrosis factor-α ( P01375 -α ) antagonist used to treat several immunologic diseases . It is believed that unexpected adverse events related to anti- P01375 drugs will occur as administration of this drug becomes more widespread . We report the case of a 37-year-old woman who complained of nonspecific abnormalities of vision in both eyes one month after beginning treatment with adalimumab for Crohn 's disease . The results of the initial ophthalmologic examination were normal . The visual fields showed diffuse and severe depression in both eyes . Other diseases that could have caused the visual field defects were ruled out . The results of electrophysiological tests were compatible with diffuse bilateral retinopathy . Treatment with adalimumab was discontinued . Six months later , visual field loss persists , although it has improved slightly . To our knowledge , this is the first report of diffuse retinopathy probable caused by systemic administration of adalimumab . This form of retinal toxicity should be considered in patients with disorders of vision treated with this drug . Use of adalimumab in patients with juvenile idiopathic arthritis refractory to etanercept and/or infliximab . To analyse the effectiveness and safety of adalimumab in a group of patients with juvenile idiopathic arthritis ( JIA ) who had failed treatment with etanercept and/or infliximab in a single paediatric rheumatology clinic . Patients with JIA with active polyarthritis refractory to metotrexate ( MTX ) ( > or =20 mg/m2/week ) for at least 3 months and to etanercept ( up to 1 mg/kg twice weekly ) and/or infliximab ( up to 10 mg/kg every 4 weeks ) for at least 6 months were included . All patients received adalimumab 24 mg/m2/week concomitantly with MTX 7.5-10 mg/week . Evaluation of efficacy included improvement as defined by the P10323 paediatric 30 criteria , 50 % and 70 % improvement and remission . Six patients were included . Three patients met improvement criteria ; 50 % and 70 % improvement occurred in two children . Improvement was sustained for 12 , 24 and 36 months , respectively . Remission occurred in one patient . DB00051 was discontinued due to lack of efficacy in three patients . No side effects were observed . DB00051 appears to be effective and safe in patients with JIA refractory to other anti- P01375 agents . Further controlled studies are needed in order to assess efficacy of adalimumab in children with refractory JIA . DB00051 ( HUMIRA ) : a review . DB00051 ( HUMIRA , Abbott Laboratories ) is a new fully human P01375 monoclonal antibody recently approved for the treatment of rheumatoid arthritis and undergoing trials for use in treating other conditions , including psoriasis and psoriatic arthritis . This article reviews its mechanisms of action , clinical trial results , and related discussion . Redo Ileal pouch-anal anastomosis combined with anti- P01375 -α maintenance therapy for Crohn 's disease with pelvic fistula : report of two cases . Pouch failure has been reported to occur after ileal pouch-anal anastomosis for Crohn 's disease . We report two cases of patients with Crohn 's disease , who underwent redo ileal pouch-anal anastomosis ( redo-IPAA ) combined with anti- P01375 -α maintenance therapy , with good functional results . The first patient , a man with presumed ulcerative colitis , suffered pelvic fistula recurrence and anastomotic dehiscence . He underwent redo-IPAA , at which time longitudinal ulcers were found . DB00065 was started 4 days postoperatively and continued . The second patient , a woman treated for ulcerative colitis , underwent laparoscopic IPAA 8 years later . After the development of a pelvic fistula , twisted mesentery of the ileal pouch was found intraoperatively and Crohn 's disease was diagnosed . DB00051 therapy resulted in fistula closure . Redo-IPAA was performed to normalize the twisted mesentery of the ileal pouch . No complications have been observed in either patient , both of whom have experienced good functional results after closure of the covering stomas . Open-label study of adalimumab in patients with ulcerative colitis including those with prior loss of response or intolerance to infliximab . BACKGROUND : The aim of this study was to assess the clinical benefit and tolerability of adalimumab , a fully human monoclonal antibody to tumor necrosis factor ( P01375 ) , in patients with ulcerative colitis ( UC ) . METHODS : Patients with active UC , including those who had lost response or developed intolerance to the chimeric anti- P01375 antibody infliximab , were enrolled in a 24-week uncontrolled trial . Patients were treated with subcutaneous adalimumab 160 mg at week 0 , 80 mg at week 2 , and 40 mg every other week starting at week 4 . After week 8 the dose could be escalated to 40 mg weekly for incomplete response . Outcome measures included clinical response and remission and mucosal healing . RESULTS : Twenty patients were enrolled , of whom 13 had previously received infliximab . Seven patients had dose escalation of adalimumab between weeks 8 and 16 , from 40 mg every other week to 40 mg weekly , due to incomplete response . The rates of clinical response were 25 % at week 8 and 50 % at week 24 . The rates of clinical remission were 5 % at week 8 and 20 % at week 24 . The rate of mucosal healing was 30 % at week 8 . The rates of clinical response and remission and mucosal healing were similar in infliximab-naïve and previously exposed patients . None of the patients experienced hypersensitivity reactions during treatment with adalimumab . CONCLUSIONS : DB00051 is well tolerated and appears to be a clinically beneficial option for patients with UC , including those who have previously lost their response to or can not tolerate infliximab . Reduced inflammation and lymphoid tissue immunopathology in rhesus macaques receiving anti-tumor necrosis factor treatment during primary simian immunodeficiency virus infection . BACKGROUND : Human immunodeficiency virus ( HIV ) and simian immunodeficiency virus ( SIV ) infections induce robust , generalized inflammatory responses that begin during acute infection and lead to pathological systemic immune activation , fibrotic damage of lymphoid tissues , and CD4⁺ T-cell loss , pathogenic processes that contribute to disease progression . METHODS : To better understand the contribution of tumor necrosis factor ( P01375 ) , a key regulator of acute inflammation , to lentiviral pathogenesis , rhesus macaques newly infected with SIVmac239 were treated for 12 weeks in a pilot study with adalimumab ( Humira ) , a human anti- P01375 monoclonal antibody . RESULTS : DB00051 did not affect plasma SIV RNA levels or measures of T-cell immune activation ( P28907 or Ki67 ) in peripheral blood or lymph node T cells . However , compared with untreated rhesus macaques , adalimumab-treated rhesus macaques showed attenuated expression of proinflammatory genes , decreased infiltration of polymorphonuclear cells into the T-cell zone of lymphoid tissues , and weaker antiinflammatory regulatory responses to SIV infection ( ie , fewer presumed alternatively activated [ ie , CD163⁺ ] macrophages , interleukin 10-producing cells , and transforming growth factor β-producing cells ) , along with reduced lymphoid tissue fibrosis and better preservation of CD4⁺ T cells . CONCLUSIONS : While HIV/SIV replication drives pathogenesis , these data emphasize the contribution of the inflammatory response to lentiviral infection to overall pathogenesis , and they suggest that early modulation of the inflammatory response may help attenuate disease progression . Long-term follow-up of adalimumab monotherapy for rheumatoid arthritis in Japanese patients : a report of six cases . We present six cases of patients with Japanese rheumatoid arthritis ( RA ) treated with a tumor necrosis factor ( P01375 ) -alpha blocking agent , adalimumab as monotherapy for 220 weeks . All six patients were women , and the median age was 54.0 ± 7.07 years old . The median duration of the disease was 7.43 ± 11.1 years , and the median disease activity score ( DAS28-CRP ) was 5.35 ± 0.69 . Three of six patients were able to continue to receive this treatment for 220 weeks successfully , and the DAS28-CRP decreased to 1.89 ± 0.75 . Two patients withdrew because of lack of efficacy , and one patient withdrew because of adverse events ( non-Hodgkin lymphoma ) . DB00051 resulted in a sustained clinical response in RA patients during 220-week follow-up . Usefulness of DB00051 for Treating a Case of Intestinal Behçet 's Disease With Trisomy 8 Myelodysplastic Syndrome . Behçet 's disease ( BD ) is a systemic vasculitis , while myelodysplastic syndrome ( P43034 ) is a heterogeneous group of clonal hematologic disorders characterized by ineffective hematopoiesis . Some studies suggest a relationship between P43034 and BD , especially intestinal BD , and trisomy 8 seems to play an important role in both diseases . There are several reports on patients with BD comorbid with P43034 involving trisomy 8 that frequently have intestinal lesions refractory to conventional medical therapies . P01375 ( P01375 ) -α is strongly involved in the pathophysiology of several autoimmune diseases such as rheumatoid arthritis , inflammatory bowel disease , and BD . In addition , P01375 -α plays an important role in the pathophysiology of P43034 by inhibiting normal hematopoiesis and inducing the programmed cell death of normal total bone marrow cells and normal P28906 + cells . Recent clinical reports demonstrate the favorable effect of P01375 -α antagonists in patients with refractory intestinal BD and in those with P43034 . We present the case of a patient with intestinal BD and P43034 involving trisomy 8 who was successfully treated with adalimumab . Monomeric IgA can be produced in planta as efficient as IgG , yet receives different N-glycans . The unique features of IgA , such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE , make it a promising antibody isotype for several therapeutic applications . However , in contrast to IgG , reports on plant production of IgA are scarce . We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana : DB00065 and DB00051 , directed against P01375 -α , and Ustekinumab , directed against the interleukin-12p40 subunit . We evaluated antibody yield , quality and N-glycosylation . All six antibodies had comparable levels of expression between 3.5 and 9 % of total soluble protein content and were shown to have neutralizing activity in a cell-based assay . However , IgA1κ-based DB00051 and Ustekinumab were poorly secreted compared to their IgG counterparts . DB00065 was poorly secreted regardless of isotype backbone . This corresponded with the observation that both IgA1κ- and IgG1κ-based DB00065 were enriched in oligomannose-type N-glycan structures . For IgG1κ-based Ustekinumab and DB00051 , the major N-glycan type was the typical plant complex N-glycan , biantennary with terminal N-acetylglucosamine , β1,2-xylose and core α1,3-fucose . In contrast , the major N-glycan on the IgA-based antibodies was xylosylated , but lacked core α1,3-fucose and one terminal N-acetylglucosamine . This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein . Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives . DB00227 -stimulated superinduction of P16581 , P05362 and P19320 in P01375 activated human vascular endothelial cells . Inhibitors of P04035 ( statins ) reveal important pharmacological effects in addition to reducing the plasma LDL cholesterol level . In the pathogenesis of arteriosclerosis , transendothelial migration of various leukocytes including monocytes is a crucial step . We , therefore , investigated the expression of P16581 , intercellular cell adhesion molecule-1 ( P05362 ) and vascular cell adhesion molecule-1 ( P19320 ) in vascular endothelial cells as influenced by lovastatin . Human umbilical vein endothelial cells ( HUVECs ) express significant amounts of selectins and cell adhesion molecules ( CAMs ) within a few hours after stimulation with P01375 . This effect is potentiated by 100-200 % when the cells are pretreated with 0.1-2.5 microM lovastatin . The lovastatin-mediated increase in the cytoplasm and at the cell surface is dose-dependent and significant at lovastatin concentrations comparable to plasma levels in patients under lovastatin treatment . The lovastatin-potentiated increase of P16581 and CAMs is correlated with a corresponding increase of selectin- and P62158 -specific mRNA . We conclude that , in vivo , statin treatment could trigger an enhanced recruitment of macrophages that might support the cholesteryl ester efflux from the arteriosclerotic plaque . Onychomycosis in patients of nail psoriasis on biologic therapy : a randomized , prospective open label study comparing DB00005 , DB00065 and DB00051 . INTRODUCTION AND OBJECTIVE : The association between patients of psoriasis on anti P01375 therapy and onychomycosis has not been explored . The aim of this study was to determine the rate of onychomycosis in patients of psoriasis with nail involvement on anti P01375 therapy . MATERIALS AND METHODS : All patients of psoriasis with nail involvement seen between February 2007 - July 2012 were examined . All the patients with negative nail scrapings for fungus were enrolled . Patients found fit for biologics after investigations were randomly divided into 3 groups ( Group A : DB00065 , Group B : DB00005 and Group C : DB00051 ) . The patients were followed up every 4 weeks for 24 weeks . Repeat nail scrapings were done at week 24 . The results were compared with controls . RESULT : In total , 315 ( 178 males and 137 females ) patients were enrolled . The mean age was 37.5 ± 11.4 years . The results for scraping for fungus at the end of 24 weeks were as follows : 33 % ( 33/100 ) in patients on DB00065 followed by 15.45 % ( 17/110 ) , 13.33 % ( 14/105 ) in patients on treatment with DB00005 and DB00051 respectively as compared to 13.89 % ( 25/180 ) among controls . Onychomycosis in association with nail psoriasis was more common in males . CONCLUSION : This study revealed statistically significant association between fungal infections of the nail in patients of psoriasis on treatment with DB00065 . DB00051 : a new tumor necrosis factor antagonist and the treatment of Crohn 's disease . Antibody-neutralization studies have implicated tumor necrosis factor-alpha ( P01375 ) and IL-12 p40 in the pathogenesis of Crohn 's disease ( CD ) . Many randomized controlled trials ( RCTs ) have demonstrated the efficacy of infliximab , a chimeric monoclonal antibody to P01375 , in inducing and maintaining disease remission in patients with moderate to severe CD , including those with draining fistulas . However , infliximab is administered intravenously and is associated with immunological reactions and development of antibodies , and hence reduced efficacy . In this article , we describe the efficacy and safety profiles of a newly available recombinant , fully humanized , subcutaneously-administered , anti- P01375 monoclonal antibody , adalimumab ( Humira ) in patients with moderate to severe CD . Many recent RCTs have shown that adalimumab is not only similar to infliximab in the mode of action , efficacy , and safety , but it has the advantages of causing less anaphylactic or immunological reactions , being administered by the subcutaneous route , less need for hospitalization , and a half-life of 2 weeks that allows every other week dosage . DB00051 has also shown some efficacy in infliximab-intolerant or resistant cases . Therefore , it represents a new horizon in the treatment of CD patients , and may reduce the number of patients who require surgical intervention . DB00051 , a fully human anti- P01375 monoclonal antibody , treatment does not influence experimental UV response in the skin of rheumatoid arthritis patients . P01375 is known to play an important role in UV-induced immunomodulation and photodamage . It plays a role in UVB-mediated induction of apoptosis and is a strong inducer of the c-Jun N-terminal kinase ( JNK ) pathway , which eventually leads to the loss of dermal collagen and elastin content . Recently chimeric anti- P01375 has been introduced as a therapy for rheumatoid arthritis . The aim of the present study was to investigate the effect of anti- P01375 treatment on UV-induced DNA damage , apoptosis , and induction of matrix metallo proteinases . Twelve patients with rheumatoid arthritis were included and irradiated with 2 Q9UQD0 broadband UVB before and after administration of 0.5 mg/kg anti- P01375 monoclonal antibody . Twenty-four hours after irradiation biopsies were taken . Frozen and paraffin sections were stained for p53 , c-Jun , phosphorylated c-Jun , sunburn cells and P03956 . No significant changes were observed in the expression of p53 and sunburn cells and P03956 content after treatment with anti- P01375 , whereas a slight but significant decrease in c-Jun and phosphorylated c-Jun expression was noted ( P = 0.0250 and P = 0.0431 , respectively ) . Our results showed no influence of anti- P01375 on UV response at therapeutic doses in patients with rheumatoid arthritis . Pharmacoeconomics of adalimumab for rheumatoid arthritis , psoriatic arthritis , ankylosing spondylitis and Crohn 's disease . DB00051 is a monoclonal antibody that inhibits P01375 , an osteogenic cytokine involved in the pathogenesis of chronic , disabling inflammatory diseases . DB00051 is indicated for rheumatoid arthritis , psoriatic arthritis , ankylosing spondylitis and Crohn 's disease . It alleviates the symptoms of these diseases , prevents disease progression in some patients and , in the case of Crohn 's disease , induces and maintains remission . Compared with traditional disease-modifying antirheumatic drugs that offer significantly less benefit , adalimumab is much more costly . However , most studies to date demonstrate the cost-effectiveness of adalimumab treatment . Cost-effectiveness data for newer indications of adalimumab , including ankylosing spondylitis and Crohn 's disease , are needed . As longer term data for adalimumab become available , the cost-effectiveness data will have greater precision . A human monoclonal anti- P01375 alpha antibody ( adalimumab ) reduces airway inflammation and ameliorates lung histology in a murine model of acute asthma . BACKGROUND : A few experimental studies related to asthma have unveiled the beneficial effects of P01375 alpha blocking agents on the airway histology , cytokine levels in bronchoalveolar lavage and bronchial hyper-responsiveness . In the current study , we aimed to assess the effect of adalimumab on the inflammation and histology of asthma in a murine model . METHOD : Twelve-week-old BALB/c ( H-2d/d ) female rats ( n=18 ) were allocated into three groups , including ( group I ) control ( phosphate-buffered saline was implemented ) , ( group II ) asthma induced with OVA ( n=6 ) , and ( group III ) asthma induced with OVA+treated with adalimumab ( n=6 ) . Rats were executed on the 28th day of the study . The lung samples were fixed in 10 % neutral buffered formalin . Lung parenchyma , alveolus , peribronchial and perivascular inflammation were assessed . Lung pathological scoring was performed . RESULT : Severity of lung damage was found to be reduced significantly in the asthma induced with OVA+treated with adalimumab group . When compared with the untreated group , adalimumab significantly reduced the inflammatory cells around the bronchi and bronchioles , and reduced inflammation of the alveolar wall and alveolar wall thickness as well ( median score=1 , p=0.52 ) . Peribronchial smooth muscle hypertrophy and oedema were significantly reduced after adalimumab administration . CONCLUSION : DB00051 ( a human monoclonal anti- P01375 alpha antibody ) therapy significantly reduced the severity of lung damage by decreasing cellular infiltration and improvement on the lung histology in a murine model of acute asthma . DB00051 for the treatment of ankylosing spondylitis . Ankylosing spondylitis is a chronic inflammatory disease , with a prevalence of approximately 0.5 % , which starts in the third decade of life . Treatment was , until recently , limited . Conventional disease-modifying drugs are not effective for the spinal manifestations , and NSAIDs and physical therapy were the standard treatment , without any other options for patients who did not respond to this treatment . Therefore , the high efficacy of the new group of P01375 -blockers for the treatment of active ankylosing spondylitis represents a breakthrough for NSAID-refractory patients . Following the introduction of the two P01375 -blockers , infliximab and etanercept , the fully humanized , anti- P01375 monoclonal antibody adalimumab is now the third product that has been approved for the treatment of ankylosing spondylitis . DB00051 is given subcutaneously every 2 weeks at a dose of 40 mg . In open and placebo-controlled trials , the drug was shown to be safe and effective in ankylosing spondylitis patients . Long-term treatment data of up to 2 years are now available , confirming efficacy and acceptable safety . DB00051 treatment in Crohn 's disease does not induce early changes in regulatory T cells . OBJECTIVE : Anti- P01375 -α antibodies has been suggested to modulate regulatory T cell ( Treg ) percentages in rheumatoid arthritis , but results from studies of Crohn 's disease ( CD ) are conflicting . We investigated dynamic changes of circulating Tregs in CD during treatment with the anti- P01375 -α-antibody adalimumab ( Humira® , Abbott Laboratories A/S , Emdrupvej 28C , DK-2100 Copenhagen ) . MATERIAL AND METHODS : Blood samples from 26 CD patients were analysed using flow cytometry before and 1 and 26 weeks after initiation of adalimumab treatment to determine the percentage of Tregs among P01730 + T cells . RESULTS : In spite of a significant decline in disease activity scores and biochemical markers of inflammation , during the first week of treatment , we did not observe early modulating effects of adalimumab on Treg percentages . However , we found a long-term increase in Treg percentages in responders who had low Treg percentages ( < 5 % ) at baseline ( p = 0.04 ) . Treg percentage was inversely associated with disease activity ( CD activity index or CDAI ) ( Spearman 's rank correlation , ρ = -0.47 , p = 0.02 ) . High Treg percentages among P01730 + T cells at baseline predicted clinical response to adalimumab . CONCLUSIONS : DB00051 treatment did not induce early modulatory effects on Treg percentage , even in responders . This finding suggests that adalimumab does not have a direct or selective effect on Tregs . However , Treg percentage was associated with disease activity and high Treg percentage predicted response to adalimumab . Refractory dissecting Cellulitis of the Scalp Successfully controlled with DB00051 . Dissecting cellulitis of the scalp ( DB00260 ) is an uncommon inflammatory disease that often results in scarring alopecia . Numerous therapies have either proved ineffective or only temporarily effective in the management of this condition . Recent reports show adequate responses to tumor necrosis factor ( P01375 ) inhibitors in cases of DB00260 . We report a case of severe recalcitrant DB00260 successfully treated with adalimumab . Microglial activation , increased P01375 and P31645 expression in the prefrontal cortex define stress-altered behaviour in mice susceptible to anhedonia . A chronic stress paradigm comprising exposure to predation , tail suspension and restraint induces a depressive syndrome in C57BL/6J mice that occurs in some , but not all , animals . Here , we sought to extend our behavioural studies to investigate how susceptibility ( sucrose preference < 65 % ) or resilience ( sucrose preference > 65 % ) to stress-induced anhedonia affects the 5HT system and the expression of inflammation-related genes . All chronically stressed animals , displayed increased level of anxiety , but susceptible mice exhibited an increased propensity to float in the forced swim test and demonstrate hyperactivity under stressful lighting conditions . These changes were not present in resilient or acutely stressed animals . Compared to resilient animals , susceptible mice showed elevated expression of tumour necrosis factor alpha ( P01375 ) and the 5-HT transporter ( P31645 ) in the pre-frontal area . Enhanced expression of 5HT(2A) and P23219 in the pre-frontal area was observed in all stressed animals . In turn , indoleamine-2,3-dioxygenase ( P14902 ) was significantly unregulated in the raphe of susceptible animals . At the cellular level , increased numbers of Iba-1-positive microglial cells were also present in the prefrontal area of susceptible animals compared to resilient animals . Consequently , the susceptible animals display a unique molecular profile when compared to resilient , but anxious , animals . Unexpectedly , this altered profile provides a rationale for exploring anti-inflammatory , and possibly , P01375 -targeted therapy for major depression . [ Differences in pharmacology of tumor necrosis factor ( P01375 ) antagonists ] . The commercially available inhibitors of P01375 are constituted by two classes of molecules : the soluble receptors ( DB00005 : Amgen Inc. Wyeth ) and the monoclonal antibodies ( DB00051 : Abbott Laboratories and DB00065 : Centocor , Inc. ) . The differences in their molecular structure , mechanism of action , pharmacokinetics ( PK ) and pharmacodynamics ( PD ) are discussed , along with the differences concerning dose , administration regimens , drug concentrations and pharmacological interactions . In order to explain the clinical differences observed when these agents are used in the " real world " , which can arise from the respective PK characteristics ( kinetics , route and frequency of administration , type of P01375 binding , effects on cytokines ) and PD responses and peculiar mechanisms of action , with distinctive immune function ( LFTα inactivation ; apoptosis induction , P01375 immunoprecipitation , C1q binding and CDC induction ; Fcγ cross-linking and ADCC induction ) , the dynamics of interaction of the two classes of neutralizing molecules with P01375 , and the ability in restoring P01375 homeostasis , are outlined . P01375 inhibitors to treat ulcerative colitis in a metastatic breast cancer patient : a case report and literature review . DB00051 ( P00813 ) is a tumor necrosis factor ( P01375 ) inhibitor , used for the treatment of inflammatory bowel disease . Previous studies have reported an increased risk of cancer following exposure to P01375 inhibitors , but little has been reported for patients with cancer receiving P01375 -inhibitor treatment . We present a female patient with metastatic breast cancer and ulcerative colitis ( UC ) who was treated with P00813 . A 54-year-old African American female with a past history of left-sided breast cancer ( BC ) diagnosed at age 30 was initially treated with left-breast lumpectomy , axillary dissection , followed by chemotherapy and radiation therapy . Years after initial diagnosis , she developed recurrent , bilateral BC and had bilateral mastectomy . Subsequent restaging computed tomography ( CT ) scan demonstrated distant metastases to the bone and lymph nodes . Three years into her treatment of metastatic breast cancer , she was diagnosed with UC by colonoscopy . Her UC was not controlled for 5 mo with 5-aminosalicylates . Subcutaneous P00813 was started and resulted in dramatic improvement of UC . Four months after starting P00813 , along with ongoing chemotherapy , restaging CT scan showed resolution of the previously seen metastatic lymph nodes . Bone scan and follow-up positron emission tomography/CT scans performed every 6 mo indicated the stability of healed metastatic bone lesions for the past 3 years on P00813 . While P01375 -α inhibitors could theoretically promote further metastases in patients with prior cancer , this is the first report of a patient with metastatic breast cancer in whom the cancer has remained stable for 3 years after P00813 initiation for UC . Serum cytokine profile in adalimumab-treated refractory uveitis patients : decreased Q9GZX6 correlates with clinical responses . PURPOSE : To report the effect of adalimumab on serum cytokines in chronic refractory uveitis . METHODS : Prospective study on the effects of adalimumab on serum cytokine levels at different time points in a cohort of 12 refractory chronic uveitis patients . Results were analyzed according to clinical outcomes and compared with systemic steroid-treated recurrent uveitis patients . RESULTS : Before treatment , patients exhibited significantly increased IL-1β , P05231 , P01375 α , IL-12 P08133 , P22301 , and Q9GZX6 . DB00051 significantly decreased P05231 , and IL-12p70 at early time points ( after 1 month of treatment ) . DB00051 effects on P22301 and Q9GZX6 appeared later ( after 6 months of treatment ) . IL-1β , Q16552 , and P01375 α were not modified at any time point . Only decreased Q9GZX6 serum levels correlated with disease activity ( p = 0.011 ) . These effects were not observed in steroid-treated patients . CONCLUSIONS : DB00051 induced drug-specific and time-dependent declines in serum levels of particular cytokines , although only Q9GZX6 correlated with disease activity in 10 out 12 patients . DB00051 : a review of its use in adult patients with rheumatoid arthritis . DB00051 ( Humira ) is a recombinant , fully human anti-tumor necrosis factor ( P01375 ) monoclonal antibody approved in the US and Europe for the treatment of adult patients with moderate to severe , active rheumatoid arthritis ( RA ) . In combination with methotrexate or standard antirheumatic therapy or as monotherapy , adalimumab effectively reduced signs and symptoms of RA , induced remission , improved physical function and inhibited the progression of structural damage in several randomized , double-blind , placebo-controlled phase III trials . The drug was generally well tolerated , with most adverse events being mild to moderate , and the serious adverse events profile being similar to that generally seen in patients with RA not receiving anti- P01375 agents . DB00051 was at least as cost effective as other anti- P01375 agents used in the therapy of RA , and provided significant improvements in patients ' health-related quality of life . Overall , adalimumab in combination with methotrexate or standard antirheumatic therapy is valuable as a first-line therapeutic option in patients with early , aggressive RA , and a second-line therapeutic option in patients with long-standing , moderate to severe RA . For the latter indication , adalimumab may also be used as monotherapy . DB00203 attenuates inflammation and oxidative stress in pelvic ganglia neurons after bilateral cavernosal nerve damage . Erectile dysfunction is a common complication for patients undergoing surgeries for prostate , bladder , and colorectal cancers , due to damage of the nerves associated with the major pelvic ganglia ( P29372 ) . Functional re-innervation of target organs depends on the capacity of the neurons to survive and switch towards a regenerative phenotype . O76074 inhibitors ( PDE5i ) have been successfully used in promoting the recovery of erectile function after cavernosal nerve damage ( BCNR ) by up-regulating the expression of neurotrophic factors in P29372 . However , little is known about the effects of PDE5i on markers of neuronal damage and oxidative stress after BCNR . This study aimed to investigate the changes in gene and protein expression profiles of inflammatory , anti-inflammatory cytokines and oxidative stress related-pathways in P29372 neurons after BCNR and subsequent treatment with sildenafil . Our results showed that BCNR in Fisher-344 rats promoted up-regulation of cytokines ( interleukin- 1 ( IL-1 ) β , P05231 , P22301 , transforming growth factor β 1 ( TGFβ1 ) , and oxidative stress factors ( DB02701 adenine dinucleotide phosphate ( NADPH ) oxidase , P05164 ( P05164 ) , inducible nitric oxide synthase ( P35228 ) , P01375 receptor superfamily member 5 ( P25942 ) that were normalized by sildenafil treatment given in the drinking water . In summary , PDE5i can attenuate the production of damaging factors and can up-regulate the expression of beneficial factors in the P29372 that may ameliorate neuropathic pain , promote neuroprotection , and favor nerve regeneration . DB00051 in recalcitrant severe psoriasis associated with atopic dermatitis . P01375 -α inhibitors may induce various cutaneous side effects including eczematous-like lesions . The management of such side effects can be challenging . Herein , we report a case of a 55-year-old man who had a flare-up and subsequent improvement of atopic dermatitis during treatment of severe psoriasis with adalimumab . DB00051 , a human anti- P01375 monoclonal antibody , outcome study for the prevention of joint damage in Japanese patients with early rheumatoid arthritis : the HOPEFUL 1 study . OBJECTIVES : To evaluate the efficacy and safety of adalimumab+methotrexate ( MTX ) in Japanese patients with early rheumatoid arthritis ( RA ) who had not previously received MTX or biologics . METHODS : This randomised , double-blind , placebo-controlled , multicentre study evaluated adalimumab 40 mg every other week+MTX 6-8 mg every week versus MTX 6-8 mg every week alone for 26 weeks in patients with RA ( ≤2-year duration ) . The primary endpoint was inhibition of radiographic progression ( change ( Δ ) from baseline in modified total Sharp score ( mTSS ) ) at week 26 . RESULTS : A total of 171 patients received adalimumab+MTX ( mean dose , 6.2±0.8 mg/week ) and 163 patients received MTX alone ( mean dose , 6.6±0.6 mg/week , p < 0.001 ) . The mean RA duration was 0.3 years and 315 ( 94.3 % ) had high disease activity ( DAS28 > 5.1 ) . DB00051 +MTX significantly inhibited radiographic progression at week 26 versus MTX alone ( ΔmTSS , 1.5±6.1 vs 2.4±3.2 , respectively ; p < 0.001 ) . Significantly more patients in the adalimumab+MTX group ( 62.0 % ) did not show radiographic progression ( ΔmTSS≤0.5 ) versus the MTX alone group ( 35.4 % ; p < 0.001 ) . Patients treated with adalimumab+MTX were significantly more likely to achieve American College of Rheumatology responses and achieve clinical remission , using various definitions , at 26 weeks versus MTX alone . Combination therapy was well tolerated , and no new safety signals were observed . CONCLUSIONS : DB00051 in combination with low-dose MTX was well tolerated and efficacious in suppressing radiographic progression and improving clinical outcomes in Japanese patients with early RA and high disease activity . Alopecia areata universalis elicited during treatment with adalimumab . DB00051 is a fully humanized recombinant anti-tumour-necrosis-factor ( P01375 ) monoclonal antibody which has been approved for rheumatoid arthritis , active ankylosing spondylitis , psoriatic arthritis and Crohn 's disease . We report a case of alopecia areata ( AA ) universalis occurring 6 months after administration of adalimumab monotherapy in a patient with a long-standing history of psoriatic arthritis and psoriasis . The diagnosis was confirmed by a scalp biopsy which showed a peribulbar infiltrate of both P01730 + and CD8+ T cells , CD1a+ dendritic cells as well as P34810 + and Q86VB7 + macrophages . In addition , immunofluorescence staining for P01375 was found in the mononuclear cell infiltrate . This case suggests a complex role of P01375 in the induction of AA . Acute renal failure from hemoglobinuric and interstitial nephritis secondary to iodine and mefenamic acid . DB00784 ingestion , usually in excess and over prolonged period is known to produce interstitial nephritis , or less commonly papillary necrosis , with acute renal failure . However , it is not dose-dependent for the induction of tubulointerstitial damage . Excess iodine ingestion is known to produce toxicity and possible death , but acute renal failure is rare . There is evidence from clinical and experimental data that iodine has toxic effect on tubular epithelial cells . Iodine has not been documented to produce red cell hemolysis and hemoglobinuria . We present a unique case of acute renal failure from hemoglobinuric and acute interstitial nephritis secondary to suicidal ingestion of potassium iodide solution and also ingestion of a few mefenamic acid tablets . These agents led to potentiation of the renal injury from hemoglobinuric tubulopathy , probably from the iodine , and renal dysfunction from alteration of renal perfusion by selective P23219 inhibition of prostaglandin production , and induction of acute interstitial nephritis from mefenamic acid , leading to acute renal failure which was reversible by hemodialysis and supportive therapy .