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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 ." ]
[ "DB00072", "DB00294", "DB00338", "DB00341", "DB00588", "DB00820", "DB02546", "DB02901", "DB04844" ]
"DB00072"
"MH_train_0"
"interacts_with DB09079?"
[ "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 ." ]
[ "DB00294", "DB00313", "DB00588", "DB00755", "DB00783", "DB02546", "DB02901", "DB06822", "DB08910" ]
"DB06822"
"MH_train_1"
"interacts_with DB00083?"
[ "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) ." ]
[ "DB00035", "DB00293", "DB00322", "DB00341", "DB00351", "DB00588", "DB00741", "DB00951", "DB01200" ]
"DB00341"
"MH_train_2"
"interacts_with DB00083?"
[ "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 ." ]
[ "DB00619", "DB00630", "DB00783", "DB00819", "DB00989", "DB01067", "DB01171", "DB02546", "DB04844" ]
"DB01171"
"MH_train_3"
"interacts_with DB06813?"
[ "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 ." ]
[ "DB00035", "DB00313", "DB00338", "DB00501", "DB00820", "DB00951", "DB01050", "DB02901", "DB08910" ]
"DB01050"
"MH_train_4"
"interacts_with DB06288?"
[ "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 ." ]
[ "DB00035", "DB00290", "DB00293", "DB00338", "DB00341", "DB00951", "DB01200", "DB01296", "DB02901" ]
"DB01200"
"MH_train_5"
"interacts_with DB00862?"
[ "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 ." ]
[ "DB00009", "DB00035", "DB00203", "DB00588", "DB00755", "DB00951", "DB02901", "DB04844", "DB06822" ]
"DB00203"
"MH_train_6"
"interacts_with DB00773?"
[ "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 ." ]
[ "DB00290", "DB00452", "DB00630", "DB00783", "DB00977", "DB01067", "DB01656", "DB06779", "DB08910" ]
"DB01656"
"MH_train_7"
"interacts_with DB01233?"
[ "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 ." ]
[ "DB00015", "DB00341", "DB00741", "DB00820", "DB01576", "DB02546", "DB02901", "DB04844", "DB04905" ]
"DB04844"
"MH_train_8"
"interacts_with DB00277?"
[ "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 ." ]
[ "DB00338", "DB00351", "DB00459", "DB00588", "DB00820", "DB00989", "DB01182", "DB01259", "DB09068" ]
"DB01182"
"MH_train_9"
"interacts_with DB01367?"
[ "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 ." ]
[ "DB00035", "DB00290", "DB00294", "DB00313", "DB00322", "DB00588", "DB00620", "DB01393", "DB04905" ]
"DB01393"
"MH_train_10"
"interacts_with DB09079?"
[ "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 ." ]
[ "DB00009", "DB00104", "DB00207", "DB00452", "DB00819", "DB01296", "DB04871", "DB04946", "DB06271" ]
"DB06271"
"MH_train_11"
"interacts_with DB05773?"
[ "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 ." ]
[ "DB00015", "DB00072", "DB00203", "DB00351", "DB00712", "DB00741", "DB00783", "DB00951", "DB01050" ]
"DB00072"
"MH_train_12"
"interacts_with DB00921?"
[ "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 ." ]
[ "DB00293", "DB00461", "DB00588", "DB00603", "DB00755", "DB00988", "DB01067", "DB01200", "DB08910" ]
"DB08910"
"MH_train_13"
"interacts_with DB00700?"
[ "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 ." ]
[ "DB00035", "DB00222", "DB00351", "DB00501", "DB00622", "DB00677", "DB00977", "DB02901", "DB06779" ]
"DB06779"
"MH_train_14"
"interacts_with DB01006?"
[ "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 ." ]
[ "DB00184", "DB00294", "DB00322", "DB00351", "DB00619", "DB00630", "DB00755", "DB06822", "DB09026" ]
"DB00184"
"MH_train_15"
"interacts_with DB01233?"
[ "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 ." ]
[ "DB00452", "DB00501", "DB00622", "DB00951", "DB00977", "DB00988", "DB00989", "DB01030", "DB01296" ]
"DB00989"
"MH_train_16"

Dataset Card for "qangaroo"

Dataset Summary

We have created two new Reading Comprehension datasets focussing on multi-hop (alias multi-step) inference.

Several pieces of information often jointly imply another fact. In multi-hop inference, a new fact is derived by combining facts via a chain of multiple steps.

Our aim is to build Reading Comprehension methods that perform multi-hop inference on text, where individual facts are spread out across different documents.

The two QAngaroo datasets provide a training and evaluation resource for such methods.

Supported Tasks and Leaderboards

More Information Needed

Languages

More Information Needed

Dataset Structure

Data Instances

masked_medhop

  • Size of downloaded dataset files: 324.10 MB
  • Size of the generated dataset: 107.41 MB
  • Total amount of disk used: 431.51 MB

An example of 'validation' looks as follows.


masked_wikihop

  • Size of downloaded dataset files: 324.10 MB
  • Size of the generated dataset: 373.82 MB
  • Total amount of disk used: 697.92 MB

An example of 'validation' looks as follows.


medhop

  • Size of downloaded dataset files: 324.10 MB
  • Size of the generated dataset: 105.30 MB
  • Total amount of disk used: 429.40 MB

An example of 'validation' looks as follows.


wikihop

  • Size of downloaded dataset files: 324.10 MB
  • Size of the generated dataset: 349.87 MB
  • Total amount of disk used: 673.97 MB

An example of 'validation' looks as follows.


Data Fields

The data fields are the same among all splits.

masked_medhop

  • query: a string feature.
  • supports: a list of string features.
  • candidates: a list of string features.
  • answer: a string feature.
  • id: a string feature.

masked_wikihop

  • query: a string feature.
  • supports: a list of string features.
  • candidates: a list of string features.
  • answer: a string feature.
  • id: a string feature.

medhop

  • query: a string feature.
  • supports: a list of string features.
  • candidates: a list of