interacts_with DB00382?
Multilocus variable-number tandem repeat analysis : a helpful tool for subtyping French Clostridium difficile PCR ribotype 027 isolates . The objective of this study was to evaluate the usefulness of multilocus variable-number tandem repeat analysis ( MLVA ) for typing and subtyping of Clostridium difficile . Sixty-eight strains were studied , including strains from PCR ribotypes 027 , 078/126 , 014/020/077 , 017 and 023 . The stability of variable-number tandem repeat ( VNTR ) loci was tested by comparing the MLVA results of two strains subcultured 11 times . After DNA extraction , seven tandem repeat loci ( A6 , P33681 , P13671 , E7 , P13726 , Q9UBA6 , Q13585 ) from published MLVA schemes were amplified by PCR and sequenced . The distance between two strains was determined by calculating the summed tandem repeat difference . Genomic diversity was evaluated by using the minimum spanning tree ( Bionumerics 5.1 software program ; Applied Maths ) . Among the 68 C. difficile isolates examined , 65 unique MLVA types were identified , suggesting a high discriminatory power . An overall good agreement was observed between MLVA types and PCR ribotypes . The stability of VNTR loci was good . MLVA could separate isolates of the hypervirulent PCR ribotype 027 clone in several clusters ; all 027 strains isolated within a hospital were grouped in a specific cluster or were placed very close to each other . Results of MLVA confirmed that strains from PCR ribotypes 078 and 126 were closely related although some were located in different branches of the tree . Similar results were observed for most strains from PCR ribotypes 014 , 020 and 077 . This highly discriminatory method is time-consuming and expensive , but is a valuable tool for subtyping of C. difficile , especially of 027 strains . DB05692 , a thrombin receptor ( P25116 ) antagonist for the prevention and treatment of atherothrombosis . DB05692 is a novel antiplatelet agent undergoing development by Schering-Plough Corp for the treatment and prevention of atherothrombosis . The compound is an orally administered himbacine analog that potently antagonizes the platelet thrombin receptor protease-activated receptor 1 ( P25116 ) , which leaves the procoagulant function of thrombin intact . In preclinical studies , DB05692 demonstrated no effect on bleed time or coagulation parameters . In both cynomolgus monkeys and humans , the compound had high bioavailability and inhibited ex vivo TRAP ( thrombin receptor-activating peptide ) -stimulated platelet aggregation in a potent and long-lasting manner . In a phase II clinical trial of patients undergoing percutaneous coronary intervention , DB05692 added to standard therapy with aspirin and clopidogrel did not increase major or minor thrombolysis in myocardial infarction bleeding , and demonstrated a trend toward decreased major adverse cardiovascular events versus placebo . At the time of publication , three phase III trials were underway to assess the efficacy and safety of DB05692 for at least 1 year in up to 35,000 patients with acute coronary syndromes or atherosclerosis . The distinct mechanism of action of DB05692 allows for cardiovascular protection without the liability of increased bleeding associated with other antiplatelet therapies . Phase III trials in high-risk patients will determine the use of DB05692 in cardiological practice . DB08901 induces apoptosis in imatinib-resistant human mast cells by dephosphorylating mutant D816V P10721 and silencing β-catenin signaling . Gain-of-function mutations of membrane receptor tyrosine kinase P10721 , especially gatekeeper D816V point mutation in P10721 , render kinase autoactivation , disease progression , and poor prognosis . D816V P10721 is found in approximately 80 % of the patients with systemic mastocytosis , and is resistant to the first and second generations of tyrosine kinase inhibitors ( TKI ) . The purpose of this investigation was aimed at exploring whether ponatinib ( DB08901 ) , a novel effective TKI against T315I Bcr-Abl , was active against D816V P10721 . We discovered that ponatinib abrogated the phosphorylation of P10721 harboring either V560G ( sensitive to imatinib ) or D816V mutation ( resistant to imatinib ) and the downstream signaling transduction . DB08901 inhibited the growth of D816V P10721 -expressing cells in culture and nude mouse xenografted tumor . DB08901 triggered apoptosis by inducing the release of cytochrome c and O95831 , downregulation of Mcl-1 . Furthermore , ponatinib abrogated the phosphorylation of β-catenin at the site Y654 , suppressed the translocation of β-catenin , and inhibited the transcription and DNA binding of TCF and the expression of its targets ( e.g. , Q9Y2T1 , c-MYC , and P24385 ) . Moreover , ponatinib was highly active against xenografted D816V P10721 tumors in nude mice and significantly prolonged the survival of mice with aggressive systemic mastocytosis or mast cell leukemia by impeding the expansion and infiltration of mast cells with imatinib-resistant D814Y P10721 . Our findings warrant a clinical trial of ponatinib in patients with systemic mastocytosis harboring D816V P10721 . Initial characterization of the glutamate-cysteine ligase modifier subunit Gclm(-/-) knockout mouse . Novel model system for a severely compromised oxidative stress response . Glutamate-cysteine ligase ( GCL ) is the rate-limiting enzyme in the DB00143 biosynthesis pathway . In higher eukaryotes , this enzyme is a heterodimer comprising a catalytic subunit ( P48506 ) and a modifier subunit ( P48507 ) , which change the catalytic characteristics of the holoenzyme . To define the cellular function of P48507 , we disrupted the mouse Gclm gene to create a null allele . Gclm(-/-) mice are viable and fertile and have no overt phenotype . In liver , lung , pancreas , erythrocytes , and plasma , however , DB00143 levels in Gclm(-/-) mice were 9-16 % of that in Gclm(+/+) littermates . DB00151 levels in Gclm(-/-) mice were 9 , 35 , and 40 % of that in Gclm(+/+) mice in kidney , pancreas , and plasma , respectively , but remained unchanged in the liver and erythrocytes . Comparing the hepatic GCL holoenzyme with P48506 in the genetic absence of P48507 , we found the latter had an approximately 2-fold increase in K(m) for glutamate and a dramatically enhanced sensitivity to DB00143 inhibition . The major decrease in DB00143 , combined with diminished GCL activity , rendered Gclm(-/-) fetal fibroblasts strikingly more sensitive to chemical oxidants such as H(2)O(2) . We conclude that the Gclm(-/-) mouse represents a model of chronic DB00143 depletion that will be very useful in evaluating the role of the P48507 subunit and DB00143 in numerous pathophysiological conditions as well as in environmental toxicity associated with oxidant insult . A Nile blue based infrared fluorescent probe : imaging tumors that over-express cyclooxygenase-2 . The first Golgi-localized cyclooxygenase-2 ( P35354 ) -specific near-infrared ( Q9Y3T9 ) fluorescent probe , Niblue- P13671 -IMC , able to detect cancer cells , was designed . Importantly , Niblue- P13671 -IMC preferentially labeled the tumors in a mouse tumor model with deep tissue penetration capacity . It may be a promising molecular tool for guiding tumor resection during surgery . Iso-intense neuroenteric cyst in the lower cervical spine treated with ventral resection and anterior fusion utilising sternal notch exposure : case report , technical note and literature review . A 36-year-old female patient with a long-standing asymptomatic lower thoracic scoliosis presented with sensory symptoms involving all limbs . Q9BWK5 scan demonstrated a rounded ventral intradural mass causing major deformity of the cervical cord at P13671 and P10643 levels . Unlike most previously reported neurenteric cysts , the Q9BWK5 signal characteristics of this mass were such that it could not be determined if it is cystic or solid , being iso-intense on T1- and hyperintense P24752 -weighted images . Resection was performed through a median corporectomy of P13671 and P10643 , the lesion being found to be a neurenteric cyst with an attachment to the anterior median fissure of the cord . Strut graft and cervical locking plate fixation from P01031 to P13671 was facilitated by extending the cervical incision into the sternal notch , with detachment of left-sided strap muscle insertion . The patient made an excellent recovery with complete resolution of neurological symptoms and solid fusion . The postoperative course was complicated by an anterior cervical P04141 collection which resolved spontaneously within 2 months . The literature regarding this rare condition and its management is reviewed . Although the majority of intraspinal neurenteric cysts are situated ventral to the cord , most reports of excision have been from a dorsal approach . Drainage and subtotal excision of neurenteric cysts have been previously advocated ; however , the recurrence rate is such that complete excision is advocated . This is facilitated by a ventral approach . A simplified method of utilising the sternal notch exposure is reported . The literature regarding the anatomical peculiarities pertinent to the sternal notch approach , and the reported literature regarding spinal neurenteric cysts is reviewed . The mouse IFN-alpha ( Ifa ) locus : correlation of physical and linkage maps by in situ hybridization . The physical location of the mouse IFN-alpha locus ( Ifa ) on chromosome 4 was defined by in situ hybridization of a cloned mouse IFN-alpha probe to metaphase spreads in which one chromosome 4 was present as part of a single metacentric chromosome , all other chromosomes being acrocentric . ( This approach greatly facilitates analysis and can be used even when it is difficult to obtain good banding ) . Using unbanded chromosomes , the grains were localized over the chromosome 4 part of the metacentric , in a region 0.61 +/- 0.07 ( SD ) of the distance from the centromere to the telomere . In Giemsa-banded spreads , the majority of the grains were in the region 4C3 ---- P13671 . Consideration of these results and of the known linkage maps for mouse and man indicates that the Galt - Aco-1 - Ifa syntenic group spans a distance of approximately 14 cM and suggests that the same group on human 9p will also occupy a similarly sized region , with P07902 proximal and IFL distal to the centromere . Immunohistochemical study of P15692 , angiopoietin 2 and their receptors in the neovascularization following microinjection of P13671 glioma cells into rat brain . BACKGROUND : Recent papers suggest that two angiogenic factors ( angiopoietin 2 and vascular endothelial growth factor ) cooperate in tumoral angiogenesis to support the growing tumor . The purpose of the present work was to demonstrate the existence of such cooperation in a longitudinal study of a brain tumor model during tumor growth by means of immunohistochemistry . MATERIALS AND METHODS : The study was performed on 31 rats bearing P13671 glioma . At different stages of tumor growth , the histological aspects were described and sections were immunostained for P15692 , Ang-2 and their receptors P17948 , P35968 and Tie-2 . Immunostaining was semi-quantitatively analyzed and the localization of immunostained cells was reported . RESULTS : Ang-2 and Tie-2 were detected in the endothelial cells of vessels surrounded by tumor cells , occuring early in our study , with immunostaining taking place from day 4 to day 24 . Immunostaining with P15692 ( on tumoral cells ) and P35968 ( on endothelial cells ) was present after 8 days of tumor growth . A clear increase of vessel density can be observed at the periphery of the tumors after 16 days of tumor growth . At that time , areas of necrosis were present in the tumor with concomitant P15692 and Ang-2 expression . CONCLUSION : The present study demonstrated cooperation between the early effect of Ang-2 and the secondary effect of P15692 to elaborate new vessels in a longitudinal study of experimental brain tumors . This study is favorable to the new model of tumor angiogenesis , with successive vessel cooption , regression and growth mediated by angiopoietins and P15692 . American DB01404 inhibits induced P35354 and NFKB activation in breast cancer cells . BACKGROUND : Epidemiologic evidence suggests reduced breast cancer mortality in users of American DB01404 ( AG ) ( Panax quinquefolium ) . We hypothesized that AG extract decreases proliferation of human breast cancer cells via an anti-inflammatory effect applicable to the prevention of breast and other cancers . MATERIAL AND METHODS : A defined lyophilized aqueous extract of AG ( LEAG ) was dissolved in DB01093 1mg/mL , and serially diluted in saline . The cell lines MDA MB 231 and MCF7 were stimulated with the phorbol ester PDBu and treated with 100-500 mcg/mL LEAG . Proliferation was measured by MDA assay . Induced P35354 expression was assayed by ELISA . Activation of NFkappaB by phosphorylation of the p65 subunit was quantified by CASE ( cellular activation of signaling ELISA ) . RESULTS : Both cell lines had reduced proliferation when treated with LEAG . PDBu stimulation of MDA MB 231 increased expression of the P35354 protein 20-fold at 48 hours ( P < 0.005 ) . P35354 protein expression remained at baseline concentrations in PDBu- treated MDA MB 231 cells exposed to 100 mcg/mL LEAG . The CASE assay showed a 4-fold increase in p65 activation 24 hours after PDBu treatment in normal medium , while phosphorylated p65 dropped below baseline in the cells treated with PDBu plus LEAG . CONCLUSION : In MDA MB 231 , P35354 was inducible with PDBu . This induced P35354 expression was blocked by 100 microgram/mL LEAG in a time course consistent with the decline in the activated p65 subunit of NFkappaB . These results provide an anti-inflammatory mechanism for a possible anti-cancer effect of American DB01404 . Endogenous expression of histamine H1 receptors functionally coupled to phosphoinositide hydrolysis in P13671 glioma cells : regulation by cyclic AMP . 1. The effects of histamine receptor agonists and antagonists on phospholipid hydrolysis in rat-derived P13671 glioma cells have been investigated . 2 . P35367 -stimulation caused a concentration-dependent increase in the accumulation of total [ 3H ] -inositol phosphates in cells prelabelled with [ 3H ] -myo-inositol . The rank order of agonist potencies was histamine ( EC50 = 24 microM ) > N alpha-methylhistamine ( EC50 = 31 microM ) > 2-thiazolylethylamine ( EC50 = 91 microM ) . 3 . The response to 0.1 mM histamine was antagonized in a concentration-dependent manner by the H1-antagonists , mepyramine ( apparent Kd = 1 nM ) and (+)-chlorpheniramine ( apparent Kd = 4 nM ) . In addition , (-)-chlorpheniramine was more than two orders of magnitude less potent than its (+)-stereoisomer . 4 . Elevation of intracellular cyclic AMP accumulation with forskolin ( 10 microM , EC50 = 0.3 microM ) , isoprenaline ( 1 microM , EC50 = 4 nM ) or rolipram ( 0.5 mM ) , significantly reduced the histamine-mediated ( 0.1 mM ) inositol phosphate response by 37 % , 43 % and 26 % respectively . In contrast , 1,9-dideoxyforskolin did not increase cyclic AMP accumulation and had no effect on the phosphoinositide response to histamine . 5 . These data indicate the presence of functionally coupled , endogenous histamine H1 receptors in P13671 glioma cells . Furthermore , the results also indicate that H1 receptor-mediated phospholipid hydrolysis is inhibited by the elevation of cyclic AMP levels in these cells . P35367 antagonism by cetirizine in isolated guinea pig tissues : influence of receptor reserve and dissociation kinetics . We characterised histamine H(1) receptor antagonism by cetirizine and its enantiomers on isolated guinea pig ileum and trachea . Competitive or mixed ( competitive and apparent noncompetitive ) antagonism profiles were observed . The order of potency was : chlorpheniramine > or =mepyramine > levocetirizine > cetirizine > or =terfenadine > loratadine > dextrocetirizine . The inhibitory effects of cetirizine , levocetirizine , terfenadine and loratadine were slowly reversible compared to those of dextrocetirizine or mepyramine . DB00341 and its enantiomers were inactive on L-type Ca(2+) channels . Reduction of the histamine H(1) receptor reserve by dibenamine in the ileum ( 100-fold higher than in the trachea ) showed a gradual change from the competitive profile of dextrocetirizine and mepyramine to a mixed profile . The present results show that cetirizine and levocetirizine are selective competitive but slowly reversible histamine H(1) receptor antagonists . Their mixed antagonism profile observed in the trachea can be explained by the small receptor reserve present in this tissue compared to the ileum and their very slow dissociation rate from the histamine H(1) receptor . DB05595 in lung cancer . DB00158 is essential for proliferating cells and folate transport pathways and folate-dependent metabolic processes show promise as targets for anti-neoplastic therapy . DB00158 receptor α ( P15328 ) , a folate transporter , is an attractive target for anti-neoplastic therapy due to its high affinity for folate , restricted range of expression in normal tissue and differential over-expression in malignant tissue . P15328 is expressed in non-small cell lung cancer , with a higher expression in adenocarcinoma compared with squamous cell carcinoma . DB05595 is a monoclonal antibody targeting P15328 which in pre-clinical studies led to cytotoxicity of P15328 -expressing cells , inhibited tumor growth in animal models and showed limited reactivity with normal tissue . In phase I/II trials , farletuzumab was well tolerated as a single-agent and in combination , without additive toxicity with chemotherapy . An ongoing phase II , double blind , placebo-controlled study is evaluating farletuzumab in patients with P15328 expressing metastatic adenocarcinoma of lung . Thrombin receptors and their antagonists : an update on the patent literature . IMPORTANCE OF THE FIELD : Thrombin plays a central role in cardiovascular inflammation . Most of the cellular responses to thrombin are mediated by cell surface protease-activated receptors ( PARs ) . Several preclinical studies indicate that PARs are potential targets for treating cardiovascular diseases such as thrombosis , atherosclerosis and restenosis . Among PARs , P25116 has emerged as an important therapeutic target . AREAS COVERED IN THIS REVIEW : This review covers recent advances in the development of thrombin receptors antagonists . It is focused on the search for P25116 antagonists as this is at the moment the most promising and attractive target . However , some early promising studies on PAR-3 and -4 antagonists are also reported . WHAT THE READER WILL GAIN : The review has been written in order to give to the reader hints and references that cover , in our opinion , the most interesting and/or promising approaches in this research field . TAKE HOME MESSAGE : Research on P25116 antagonists has finally led to good clinical candidates such as DB05692 ( Schering-Plough ) and E-5555 ( Eisai Co. ) . Clinical trials clearly demonstrate that development of PAR1 antagonists is not only possible but most likely will lead to development of antiplatelet drugs as well as of drugs useful for the treatment of inflammatory , proliferative and neurodegenerative diseases . Generation and phenotypic characterization of new human ovarian cancer cell lines with the identification of antigens potentially recognizable by HLA-restricted cytotoxic T cells . This study describes a simple method for long-term establishment of human ovarian tumor lines and prediction of T-cell epitopes that could be potentially useful in the generation of tumor-specific cytotoxic T lymphocytes ( CTLs ) . Nine ovarian tumor lines ( INT.Ov ) were generated from solid primary or metastatic tumors as well as from ascitic fluid . Notably all lines expressed HLA class I , intercellular adhesion molecule-1 ( P05362 ) , polymorphic epithelial mucin ( P15941 ) and cytokeratin ( CK ) , but not HLA class II , P33681 .1 ( P33681 ) or Q13072 . While of the 9 lines tested 4 ( INT.Ov1 , 2 , 5 and 6 ) expressed the folate receptor ( P15328 ) and 6 ( INT.Ov1 , 2 , 5 , 6 , 7 and 9 ) expressed the epidermal growth factor receptor ( P00533 ) ; MAGE-1 and p185HER-2/neu were only found in 2 lines ( INT.Ov1 and 2 ) and Q13065 expression in 1 line ( INT.Ov2 ) . The identification of class I MHC ligands and T-cell epitopes within protein antigens was achieved by applying several theoretical methods including : 1 ) similarity or homology searches to MHCPEP ; 2 ) BIMAS and 3 ) artificial neural network-based predictions of proteins MAGE , GAGE , P00533 , p185HER-2/neu and P15328 expressed in INT.Ov lines . Because of the high frequency of expression of some of these proteins in ovarian cancer and the ability to determine HLA binding peptides efficiently , it is expected that after appropriate screening , a large cohort of ovarian cancer patients may become candidates to receive peptide-based vaccines . Characterization of the rat Q99259 gene promoter reveals elements important for basal transcription and glucose responsiveness . Q05329 and Q99259 are two isoforms of the enzyme glutamic acid decarboxylase which catalyze the production of GABA from glutamate , primarily in the brain . However , Q99259 and GABA also prevail in the retina , testes and islets of Langerhans . The main function of GABA is in neurotransmission , and it is involved in paracrine signalling in islets , but has also been suggested to play a role as a trophic factor in synaptogenesis and to be an important metabolite feeding into the tricarboxylic acid cycle via the GABA-shunt . Both Q99259 isoforms are subject to regulation , e.g. by synaptic activity . Q05329 is regulated at the level of enzyme activity by association and dissociation from its cofactor , PLP , whereas Q99259 is controlled at the level of its mRNA . To study this process in further detail , we have isolated and characterized the 5'-flanking region of the rat Q99259 gene . We report the transcriptional initiation sites and promoter sequences important for expression in islet beta-cells and P13671 glioma cells , and demonstrate that the Q99259 promoter harbors elements that are responsive to glucose in primary islet cells . DB00114 values in cerebrospinal fluid : reference values and diagnosis of Q9NVS9 deficiency in paediatric patients . Our aim was to establish reference values for cerebrospinal fluid ( P04141 ) pyridoxal 5'-phosphate ( PLP ) in a paediatric population for the diagnosis of pyridox(am)ine 5'-phosphate oxidase ( Q9NVS9 ) deficiency . For reference values , P04141 samples from 113 paediatric controls ( age range : 1 day-18 years ) from Barcelona and London were analysed . Cerebrospinal fluid PLP and biogenic amine concentrations were analysed by HPLC with fluorescence and electrochemical detection . DB00114 concentrations in 4 patients with Q9NVS9 deficiency were determined . A negative correlation between P04141 PLP values and age of controls was observed in both populations ( r=-0.503 ; p < 0.0001 and r=-0.542 ; p=0.002 ) . Reference values were stratified into 4 ( Barcelona ) and 3 age groups ( London ) . For the newborn period , P04141 PLP reference intervals were 32-78 and 44-89 nmol/L for the Barcelona and London centers , respectively ) . No correlation was observed in the different age groups between PLP values and biogenic amines metabolites . PLP values in neonates with Q9NVS9 deficiency were clearly decreased ( PLP=3.6 , 12.0 , 14.0 and 18.0 nmol/L ) compared with our reference ranges . In conclusion , reference values for P04141 PLP should be stratified according to age . No association was observed between PLP values and biogenic amines metabolites . In our 4 cases with Q9NVS9 deficiency , P04141 PLP values were clearly below the reference values . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . DB05595 , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC(50) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 did not block membrane binding of radiolabeled folic acid , 5-methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 had a minimal effect on the cytoplasmic accumulation of 5-methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC(50) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated . DB02342 in the human corpus luteum throughout the luteal phase and its influence on lutein cell steroidogenesis and angiogenic activity . OBJECTIVE : To quantitate 2-methoxyestradiol ( 2-ME ) in human corpus luteum ( CL ) of different ages and to determine the expression of cytochrome-P450-1A1 ( P04798 ) and catechol-O-methyl transferase ( P21964 ) in CL and the action of 2-ME on P , vascular endothelial growth factor ( P15692 ) secretion , and luteal angiogenesis . DESIGN : Experimental study . SETTING : University division of reproductive endocrinology . PATIENT(S) : Twenty-four women of reproductive age . INTERVENTION(S) : CL was collected from 15 women during the minilaparotomy for tubal sterilization . Granulosa lutein cells were harvested 36 hours after hCG administration in patients undergoing IVF . MAIN OUTCOMES MEASURE(S) : Levels of 2-ME were determined by high-performance liquid chromatography in CL . P04798 and P21964 were assessed by immunohistochemistry and Western blot . P and P15692 were measured by radioimmunoassay and ELISA . The angiogenic potential was analyzed using EA.hy926 cells . RESULT(S) : Plasma levels of E₂ decreased in the late luteal phase in association with an increase in luteal tissue of 2-ME concentrations . Concomitantly , there was a significant reduction of angiogenic activity in late CL . There was no significant variation in P04798 and P21964 expression in all CL . In physiological doses , 2-ME inhibited basal P15692 by granulosa lutein cells and diminished the angiogenic activity in conditioned media but did not prevent P and P15692 production stimulated by hCG . CONCLUSION(S) : These data suggest the participation of 2-ME in physiological luteolysis by reducing angiogenesis . However , 2-ME did not prevent in vitro hCG stimulation of P biosynthesis , providing a mechanism for CL rescue in the cycle of conception . NFκB inhibitors induce cell death in glioblastomas . Identification of novel target pathways in glioblastoma ( GBM ) remains critical due to poor prognosis , inefficient therapies and recurrence associated with these tumors . In this work , we evaluated the role of nuclear-factor-kappa-B ( NFκB ) in the growth of GBM cells , and the potential of NFκB inhibitors as antiglioma agents . NFκB pathway was found overstimulated in GBM cell lines and in tumor specimens compared to normal astrocytes and healthy brain tissues , respectively . Treatment of a panel of established GBM cell lines ( U138MG , U87 , U373 and P13671 ) with pharmacological NFκB inhibitors ( BAY117082 , parthenolide , MG132 , curcumin and arsenic trioxide ) and NFκB-p65 siRNA markedly decreased the viability of GBMs as compared to inhibitors of other signaling pathways such as MAPKs ( P29323 , JNK and p38 ) , PKC , P00533 and PI3K/Akt . In addition , NFκB inhibitors presented a low toxicity to normal astrocytes , indicating selectivity to cancerous cells . In GBMs , mitochondrial dysfunction ( membrane depolarization , bcl-xL downregulation and cytochrome c release ) and arrest in the G2/M phase were observed at the early steps of NFκB inhibitors treatment . These events preceded sub- P55008 detection , apoptotic body formation and caspase-3 activation . Also , NFκB was found overstimulated in cisplatin-resistant P13671 cells , and treatment of GBMs with NFκB inhibitors overcame cisplatin resistance besides potentiating the effects of the chemotherapeutics , cisplatin and doxorubicin . These findings support NFκB as a potential target to cell death induction in GBMs , and that the NFκB inhibitors may be considered for in vivo testing on animal models and possibly on GBM therapy . 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 . Effect of tumour necrosis factor-alpha on estrogen metabolic pathways in breast cancer cells . P01375 -alpha ( P01375 -α ) is a proinflammatory cytokine that has been linked to breast cancer development . Estrogen metabolic pathway is also involved in breast carcinogenesis and DNA adducts formation . In this study we investigated the effect of P01375 -α on the estrogen metabolic pathway in MCF-7 , a breast cancer cell line . Capillary liquid chromatography/mass spectrometry ( LC/MS ) and High performance liquid chromatography ( HPLC ) were used for analysis of estrogen metabolites and estrogen-DNA adducts levels respectively . Reporter gene assay , Real time reverse transcription polymerase chain reaction ( real time RT-PCR ) and Western blot were used to assess the expression of estrogen metabolizing genes and enzymes . P01375 -α significantly increased the total EM and decreased the estrone ( E1 ) / 17-β estradiol ( E2 ) ratio . Moreover , it altered the expression of genes and enzymes involved in E2 activation and deactivation pathways e.g. Cytochrome P-450 1A1 ( P04798 ) , Cytochrome P-450 1B1 ( Q16678 ) , Catechol-O-methyl transferase ( P21964 ) and DB02701 adenine dinucleotide phosphate-quinone oxidoreductase 1 ( P15559 ) . In addition , there were increased levels of some catechol estrogens e.g. 4-hydroxy-estrone ( 4-OHE1 ) and 2-hydroxyestradiol ( 2-OHE2 ) with decreased levels of methylated catechols e.g. 2-methoxy estradiol ( DB02342 ) . DNA adducts especially 4-OHE1-[2]-1-N3 DB00173 was significantly increased . P01375 -α directs the estrogen metabolism into more hormonally active and carcinogenic products in MCF-7 . This may implicate a new possible explanation for inflammation associated breast cancer . Subcellular localization of galactose-1-phosphate uridylyltransferase in the yeast Saccharomyces cerevisiae . The enzyme galactose-1-phosphate uridylyltransferase ( P07902 ) catalyzes the second step of the Leloir pathway of galactose metabolism , following galactokinase ( P51570 ) and preceding DB03501 -4-epimerase ( Q14376 ) . Impairment of P07902 in humans results in the potentially lethal disorder classic galactosemia . Standard lysis protocols of bacteria , yeast , or mammalian cells release all three Leloir enzymes in the soluble fraction , leading to the historical assumption that all three function as free cytosolic enzymes . We have tested this assumption with regard to P07902 in vivo using the yeast Saccharomyces cerevisiae , by linking a GFP-tag onto the amino terminus of Gal7p , the endogenous yeast P07902 . We find clear evidence of localization of the fusion protein to discrete spots in the cytoplasm of the majority of cells expressing all three Leloir enzymes , although GFP alone appears freely cytosolic . In contrast , yeast expressing GFP-Gal7p but lacking Gal1p ( P51570 ) , Gal10p ( Q14376 ) , or both do not demonstrate spots in the majority of cells , implicating a role , either direct or indirect , for these other Leloir proteins in the Gal7p localization process . Preliminary truncation experiments reveal that amino acids 1-134 of Gal7p are sufficient to drive localization of the fusion protein , while amino acids 1-66 are not . Finally , GFP-tagged human P07902 expressed in yeast also localizes to spots , demonstrating that at least some of the intrinsic determinants of localization have been conserved . These observations raise the intriguing possibility that P07902 may function in a sequestered rather than a freely diffusible state , and that this subcellular organization may have been conserved through evolution . Amelioration of scopolamine induced cognitive dysfunction and oxidative stress by Inonotus obliquus - a medicinal mushroom . The present study was aimed to investigate the cognitive enhancing and anti-oxidant activities of Inonotus obliquus ( Chaga ) against scopolamine-induced experimental amnesia . Methanolic extract of Chaga ( Q9NRJ3 ) at 50 and 100 mg kg (-1)doses were administered orally for 7 days to amnesic mice . Learning and memory was assessed by passive avoidance task ( PAT ) and Morris water maze ( MWM ) test . Tacrine ( DB00382 , 10 mg kg ( -1 ) , orally ( p.o ) ) used as a reference drug . To elucidate the mechanism of the cognitive enhancing activity of Q9NRJ3 , the activities of acetylcholinesterase ( P22303 ) , anti-oxidant enzymes , the levels of acetylcholine ( ACh ) and nitrite of mice brain homogenates were evaluated . Q9NRJ3 treatment for 7 days significantly improved the learning and memory as measured by PAT and MWM paradigms . Further , Q9NRJ3 significantly reduced the oxidative-nitritive stress , as evidenced by a decrease in malondialdehyde and nitrite levels and restored the glutathione and superoxide dismutase levels in a dose dependent manner . In addition , Q9NRJ3 treatment significantly decreased the P22303 activity in both the salt and detergent-soluble fraction of brain homogenates . Further , treatment with Q9NRJ3 restored the levels of ACh as did DB00382 . Thus , the significant cognitive enhancement observed in mice after Q9NRJ3 administration is closely related to higher brain anti-oxidant properties and inhibition of P22303 activity . These findings stress the critical impact of Chaga , a medicinal mushroom , on the higher brain functions like learning and memory . Combined environmental risk assessment for 5-fluorouracil and capecitabine in Europe . An environmental risk assessment ( ERA ) was made for the old cytostatic active pharmaceutical ingredient 5-fluorouracil ( DB00544 ) and for capecitabine ( CAP ) , which is a prodrug of DB00544 . This ERA is based on published and company internal data as well as new test results for physicochemical , human metabolism , biodegradability , environmental partitioning and fate , and acute and chronic ecotoxicity properties of the active substance DB00544 as well as on use sales data for DB00544 and CAP in Europe . Predicted environmental concentrations ( PECs ) were extrapolated following the EMEA 2006 Guideline on ERA for human pharmaceuticals and the European Union 2003 Technical Guidance Document ( TGD ) for risk assessment as well as the TGD-based application EUSES v2.0 . Actual amounts sold were taken from IMS Health Databases , in order to refine the default use and EMEA penetration factor as well as the PECs . Moreover , available measured environmental concentrations ( MECs ) were used to supplement PECs . A predicted no-effect concentration ( PNEC ) for DB00544 was derived from chronic ecotoxicity data . Except for the simplistic EMEA Phase I default PEC , the risk characterization by PEC:PNEC and Q9NRJ3 :PNEC ratios for various environmental compartments resulted in no significant risk . As the EMEA Phase I PEC does not integrate documented human metabolism and environmental degradation , in contrast to refined PEC derivations , it is inferred that the current use of CAP and DB00544 does not present any evident risk to the environment . An additional evaluation of persistence , bioaccumulation , and toxicity ( P10721 ) properties supports the conclusion of no significant environmental risk for DB00544 and CAP . DB01404 saponin metabolite induces apoptosis in MCF-7 breast cancer cells through the modulation of AMP-activated protein kinase . Previous studies have shown that the ginseng saponin metabolite , Compound K ( 20-O-d-glucopyranosyl-20(S)-protopanaxadiol , IH901 ) , suppresses proliferation of various cancers and induces apoptosis . AMP-activated protein kinase ( AMPK ) is a sensor of cellular energy states and is involved in apoptosis of cancer cells . We hypothesized that Compound K may exert cytotoxicity in MCF-7 human breast cancer cells through modulation of AMPK , followed by a decrease in cyclooxygenase-2 ( P35354 ) expression . Compound K inhibited cell growth , induced apoptosis via generation of reactive oxygen species ( ROS ) , as well as decreasing P35354 expression and prostaglandin E(2) ( PGE(2) ) levels . These effects of Compound K were induced via an AMPK-dependent pathway and were abrogated by a specific AMPK inhibitor . These results suggest that Compound K induced apoptosis by modulating AMPK- P35354 signaling in MCF-7 human breast cancer cells . Investigation of P25116 -type thrombin receptors in rat glioma P13671 cells with a novel monoclonal anti- P25116 antibody ( Mab COR7-6H9 ) . Rat glioma P13671 cells have been demonstrated to be a suitable model in the investigation of P25116 -type thrombin receptors in brain . However , anti- P25116 antibodies , which should be very helpful tools in studying P25116 in rat cells , have not been available up until now . Therefore , we prepared a monoclonal anti-thrombin receptor antibody ( Mab COR7-6H9 ) directed against the peptide sequence GRAVYLNKSRFPPMPPPPFISEDASG in the N-terminus below the thrombin cleavage site of the rat P25116 -type thrombin receptor . Using this antibody , we demonstrated the presence of P25116 binding sites on the plasma membrane of rat glioma P13671 cells both with confocal laser fluorescence and with scanning electron microscopy . In addition , Mab COR7-6H9 was shown to block P25116 -mediated transmembranal signaling as demonstrated by measurement of free intracellular calcium and cyclic AMP . This novel anti- P25116 antibody is therefore likely to be a very helpful tool in studying P25116 -type thrombin receptors in rat brain . Distinct roles of galactose-1P in galactose-mediated growth arrest of yeast deficient in galactose-1P uridylyltransferase ( P07902 ) and DB03501 4'-epimerase ( Q14376 ) . Galactose is metabolized in humans and other species by the three-enzyme Leloir pathway comprised of galactokinase ( P51570 ) , galactose 1-P uridylyltransferase ( P07902 ) , and DB03501 4'-epimerase ( Q14376 ) . Impairment of P07902 or Q14376 in humans results in the potentially lethal disorder galactosemia , and loss of either enzyme in yeast results in galactose-dependent growth arrest of cultures despite the availability of an alternate carbon source . In contrast , loss of P51570 in humans is not life-threatening , and in yeast has no impact on the growth of cultures challenged with galactose . Further , the growth of both P07902 -null and Q14376 -null yeast challenged with galactose is rescued by loss of P51570 , thereby implicating the P51570 reaction product , gal-1P , for a role in the galactose-sensitivity of both strains . However , the nature of that relationship has remained unclear . Here we have developed and applied a doxycycline-repressible allele of galactokinase to define the quantitative relationship between galactokinase activity , gal-1P accumulation , and growth arrest of galactose-challenged P07902 or Q14376 -deficient yeast . Our results demonstrate a clear threshold relationship between gal-1P accumulation and galactose-mediated growth arrest in both P07902 -null and Q14376 -null yeast , however , the threshold for the two strains is distinct . Further , we tested the galactose-sensitivity of yeast double-null for P07902 and Q14376 , and found that although loss of P07902 barely changed accumulation of gal-1P , it significantly lowered the accumulation of DB03501 , and also dramatically rescued growth of the Q14376 -null cells . Together , these data suggest that while gal-1P alone may account for the galactose-sensitivity of P07902 -null cells , other factors , likely to include DB03501 accumulation , must contribute to the galactose-sensitivity of Q14376 -null cells . Impairment of nitrergic system and delayed gastric emptying in low density lipoprotein receptor deficient female mice . BACKGROUND : In the current study , we have investigated whether low density lipoprotein receptor knockout mice ( P01130 -KO ) , moderate oxidative stress model and cholesteremia burden display gastroparesis and if so whether nitrergic system is involved in this setting . In addition , we have investigated if sepiapterin ( O43157 ) supplementation attenuated impaired nitrergic system and delayed gastric emptying . METHODS : Gastric emptying and nitrergic relaxation were measured in overnight fasting mice . nNOSα dimerization , anti-oxidant markers such as Nrf2 , P48507 , P48506 , P09601 , catalase ( CAT ) , and superoxide dismutase ( P00441 ) were measured using standard methods . DB03886 levels and intestinal transit time were measured using HPLC and dye migration assay , respectively . Wild type ( WT ) and P01130 -KO were supplemented with O43157 . KEY RESULTS : In P01130 null stomachs : ( i ) significant reduction in rate of gastric emptying , gastric pyloric and fundus nitrergic relaxation and nNOSα dimerization , ( ii ) elevated oxidized biopterins and reduced ratio of BH(4) /BH(2) + B , ( iii ) reduced Nrf2 and P48506 protein expression and no change in P48507 , P09601 , CAT , P00441 , and ( iv ) accelerated small intestinal motility were noticed . Supplementation of O43157 restored delayed gastric emptying , impaired pyloric and fundus nitrergic relaxation with restoration of P29475 dimerization and P29475 expression . CONCLUSIONS & INFERENCES : This novel data suggests that hyperlipidemia and/or suppression of selective antioxidants may be a potential cause of developing gastroparesis in diabetic patients . Intraspinal sarcoidosis : clinical features , MR imaging and electrophysiological study . We presented the case of a 58-year-old woman with spinal sarcoidosis at the C4- P13671 level . The lesion itself as visualized with magnetic resonance imaging , and its neurological findings responded favorably to treatment with prednisolone . Comparison of pre- and post-treatment values for somatosensory evoked potential ( O43157 ) and central motor conduction time ( CMCT ) also showed improvement . Analysis of O43157 and CMCT can be a useful tool for following up spinal sarcoidosis . Cinnamon polyphenols attenuate the hydrogen peroxide-induced down regulation of S100β secretion by regulating sirtuin 1 in P13671 rat glioma cells . AIMS : It is well established that the brain is particularly susceptible to oxidative damage due to its high consumption of oxygen . The objective of this study was to investigate the protective effects of a water soluble polyphenol-rich extract of cinnamon and the possible mechanisms , under conditions of oxidative stress-induced by hydrogen peroxide , in rat P13671 glioma cells . MAIN METHODS : After 24h of H2O2 incubation , the secretion and intracellular expression of S100β were determined by immunoprecitation/immunoblotting and immunofluorescence imaging . KEY FINDINGS : Cinnamon polyphenols ( CP ) counteracted the oxidative effects of H2O2 on S100β secretion and expression . CP also enhanced the impaired protein levels of sirtuins 1 , 2 , and 3 , which are deacetylases important in cell survival . H2O2 also induced the overexpression of the proinflammatory factors , P01375 -α , phospho-NF-κB p65 , as well as of Bcl-xl , Bax and P42574 , which are all the members of the Bcl-2 family . CP not only suppressed the expression of these proteins but also attenuated the phosphorylation induced by H2O2 . CP also upregulated the decreased Bcl-2 protein levels in H2O2 treated P13671 cells . The effects of CP on H2O2-induced downregulation of S100β secretion were blocked by Q96EB6 siRNA demonstrating that Q96EB6 plays a regulatory role in CP-mediated prevention by H2O2 . SIGNIFICANCE : These data demonstrate that Cinnamon polyphenols may exert neuroprotective effects in glial cells by the regulation of Bcl-2 family members and enhancing Q96EB6 expression during oxidative stress . P42574 -Dependent Cleavage of the Glutamate- DB00151 Ligase Catalytic Subunit during Apoptotic Cell Death . Apoptotic cell death is usually accompanied by activation of a family of cysteine proteases termed caspases . Caspases mediate the selective proteolysis of multiple cellular targets often resulting in the disruption of survival pathways . Intracellular levels of the antioxidant glutathione ( DB00143 ) are an important determinant of cellular susceptibility to apoptosis . The rate-limiting step in DB00143 biosynthesis is mediated by glutamate-L-cysteine ligase ( GCL ) , a heterodimeric enzyme consisting of a catalytic ( P48506 ) and a modifier ( P48507 ) subunit . In this report we demonstrate that P48506 is a direct target for caspase-mediated cleavage in multiple models of apoptotic cell death . Mutational analysis revealed that caspase-mediated cleavage of P48506 occurs at DB00128 (499) within the sequence AVVD(499)G . P48506 cleavage occurs upstream of DB00151 (553) , which is thought to be important for association with P48507 . P48506 cleavage is accompanied by a rapid loss of intracellular DB00143 due to caspase-mediated extrusion of DB00143 from the cell . However , while P48506 cleavage is dependent on caspase-3 , DB00143 extrusion occurs by a caspase-3-independent mechanism . Our identification of P48506 as a target for caspase-3-dependent cleavage during apoptotic cell death suggests that this post-translational modification may represent a novel mechanism for regulating DB00143 biosynthesis during apoptosis . DB08901 ( DB08901 ) , a multitargeted pan-FGFR inhibitor with activity in multiple FGFR-amplified or mutated cancer models . Members of the fibroblast growth factor receptor family of kinases ( P11362 -4 ) are dysregulated in multiple cancers . DB08901 ( DB08901 ) is an oral multitargeted tyrosine kinase inhibitor being explored in a pivotal phase II trial in patients with chronic myelogenous leukemia due to its potent activity against P11274 - P00519 . DB08901 has also been shown to inhibit the in vitro kinase activity of all four FGFRs , prompting us to examine its potential as an FGFR inhibitor . In Ba/ P13726 cells engineered to express activated P11362 -4 , ponatinib potently inhibited FGFR-mediated signaling and viability with IC(50) values < 40 nmol/L , with substantial selectivity over parental Ba/ P13726 cells . In a panel of 14 cell lines representing multiple tumor types ( endometrial , bladder , gastric , breast , lung , and colon ) and containing FGFRs dysregulated by a variety of mechanisms , ponatinib inhibited FGFR-mediated signaling with IC(50) values < 40 nmol/L and inhibited cell growth with GI(50) ( concentration needed to reduce the growth of treated cells to half that of untreated cells ) values of 7 to 181 nmol/L . Daily oral dosing of ponatinib ( 10-30 mg/kg ) to mice reduced tumor growth and inhibited signaling in all three tumor models examined . Importantly , the potency of ponatinib in these models is similar to that previously observed in P11274 - P00519 -driven models and plasma levels of ponatinib that exceed the IC(50) values for P11362 -4 inhibition can be sustained in patients . These results show that ponatinib is a potent pan-FGFR inhibitor and provide strong rationale for its evaluation in patients with FGFR-driven cancers . Inorganic lead enhances cytokine-induced elevation of matrix metalloproteinase P14780 expression in glial cells . Inorganic lead ( Pb ) is a ubiquitous environmental contaminant that produces a variety of deleterious effects in the central nervous system ( CNS ) . Matrix metalloproteinases ( MMPs ) , specifically P14780 , induced by inflammatory cytokines , are increasingly being implicated in CNS pathology . The present study demonstrates that low concentrations of either pro-inflammatory cytokines ( P01375 and IL-1beta ) or Pb did not influence the P14780 expression in a glial cell line ( P13671 ) when added separately . However , combined administration of Pb and cytokines induced a marked synergized elevation of P14780 expression in spite of a reduction in the number of glial cells . These results demonstrate a possible new mechanism by which Pb may induce neuropathological processes . DB00114 ( PLP ) deficiency might contribute to the onset of type I diabetes . The incidence of type I diabetes is rising worldwide , particularly in young children . Type I diabetes is considered a multifactorial disease with genetic predisposition and environmental factors participating . Currently , despite years of research , there is no consensus regarding the factors that initiate the autoimmune response . Type I diabetes is preceded by autoimmunity to islet antigens , among them the protein glutamic acid decarboxylase , Q05329 . DB00114 ( PLP ) is formed from vitamin B6 by the action of pyridoxal kinase . Interaction of Q05329 with PLP is necessary for Q05329 -mediated synthesis of the neurotransmitter γ-aminobutyric acid ( GABA ) . PLP is also a required cofactor for dopamine synthesis by L-aromatic decarboxylase ( L- P20711 ) . Both Q05329 and L- P20711 are expressed in pancreatic islets . Here it is proposed that lack of the vitamin B6 derivative pyridoxal 5'-phosphate might contribute to the appearance of pancreatic islet autoimmunity and type I diabetes onset .