MH_dev_207

ABT - 737 is highly effective against molecular subgroups of multiple myeloma . Multiple myeloma is a plasma cell malignancy that is heterogeneous with respect to its causative molecular abnormalities and the treatment response of patients . The Bcl - 2 protein family is critical for myeloma cell survival . ABT - 737 is a cell-permeant compound that binds to Bcl - 2 and Bcl-x ( L ) but not to Mcl - 1 . Using a myeloma cell line collection ( n = 25 ) representative of different molecular translocations , we showed that ABT - 737 effectively kills a subset of cell lines ( n = 6 ) , with a median lethal dose ranging from 7 ± 0.4 nM to 150 ± 7.5 nM . Of interest , all sensitive cell lines harbored a t ( 11 ; 14 ) . We demonstrated that ABT - 737 - sensitive and ABT - 737 - resistant cell lines could be differentiated by the P10415 REA / Q07820 REA expression ratio . A screen of a public expression database of myeloma patients indicates that the P10415 REA / Q07820 REA ratio of t ( 11 ; 14 ) and hyperdiploid patients was significantly higher than in all other groups ( P < . 001 ) . ABT - 737 first induced the disruption of Bcl - 2 / Bax , Bcl - 2 / Bik , or Bcl - 2 / Puma complexes , followed by the disruption of Bcl - 2 heterodimers with Bak and Bim . Altogether , the identification of a subset of cell lines and primary cells effectively killed by ABT - 737 alone supported the evaluation of DB05764 MEN , an orally active counterpart to ABT - 737 , for the treatment of t ( 11 ; 14 ) and hyperdiploid groups of myeloma harboring a Bcl - 2 ( high ) / Mcl - 1 ( low ) profile .

Inhibition of the invasion capacity of carcinoma cells by DB05476 MEN , a novel synthetic inhibitor of the urokinase-type plasminogen activator system . The overall survival rate of patients suffering from carcinomas has remained poor and nearly unchanged over the last decades . This is mainly due to the so-called minimal residual disease , i . e . , remaining tumor cells that overcome surgery and / or radiotherapy and are the cause of locoregional and distant metastases . To metastasize , tumor cells take advantage of proteases to invade and remodel surrounding tissues . Here , we analyzed the efficiency of DB05476 MEN , a novel 3 - amidinophenylalanine-based inhibitor of the uPA system , at inhibiting the invasive capacity of carcinoma cells . First , Q03405 REA expression was characterized in different carcinoma cell lines , including SCCHN , breast and cervical carcinoma . Thereafter , the invasive potential of these cell lines was determined using Matrigel invasion chambers and a spheroid cocultivation model with human fibroblasts . Q03405 REA expression levels correlated positively with invasion capacity , which could be significantly inhibited by DB05476 MEN . A decrease of tumor cell invasion by up to 50 % was achieved in both models with the SCCHN line FaDu and the cervical carcinoma line HeLa after treatment with DB05476 MEN . Thus , our results demonstrate the potential of DB05476 MEN in vitro as a promising adjuvant antimetastatic therapy of carcinomas .

An overview of phenserine tartrate , a novel acetylcholinesterase inhibitor for the treatment of Alzheimer ' s disease . Existing cholinesterase ( ChE ) inhibitor therapies for Alzheimer ' s disease ( AD ) , while effective in improving cognitive , behavioral and functional impairments , do not alter disease progression . Novel drug design studies have focused on the classical ChE inhibitor , ( - ) - physostigmine , producing alterations in chemical composition and three-dimensional structure , which may offer an improved therapeutic index . The phenylcarbamate derivative , DB04892 MEN , is a selective , non-competitive inhibitor of acetylcholinesterase ( P22303 REA ) . In vivo , DB04892 MEN produces rapid , potent , and long-lasting P22303 REA inhibition . As a possible result of its preferential brain selectivity , DB04892 MEN is significantly less toxic than ( - ) - physostigmine . In studies using the Stone maze paradigm , DB04892 MEN has been shown to improve cognitive performance in both young learning-impaired and elderly rats . In addition to reducing inactivation of acetylcholine in the brain , DB04892 MEN appears to have a second mode of action . Reduced secretion of beta-amyloid ( Abeta ) has been observed in cell lines exposed to DB04892 MEN , occurring through translational regulation of beta-amyloid precursor protein ( beta - P05067 REA ) mRNA via a non-cholinergic mechanism . These in vitro findings appear to translate in vivo into animal models and humans . In a small study of patients with AD , DB04892 MEN treatment tended to reduce beta - P05067 REA and Abeta levels in plasma samples . Clinical studies also reveal that DB04892 MEN ( 5-10 mg b . i . d . ) had a favorable safety and pharmacological profile , produced significant improvements in cognitive function and was well tolerated in patients with AD treated for 12 weeks . Further randomized , double-blind , placebo-controlled Phase III studies assessing the efficacy , safety / tolerability and potential disease-modifying effects of DB04892 MEN in patients with AD are currently ongoing .

Clinical drug-drug interaction assessment of ivacaftor as a potential inhibitor of cytochrome P450 and P-glycoprotein . DB08820 MEN is approved in the USA for the treatment of cystic fibrosis ( CF ) in patients with a G551D - P13569 REA mutation or one of eight other P13569 REA mutations . A series of in vitro experiments conducted early in the development of ivacaftor indicated ivacaftor and metabolites may have the potential to inhibit cytochrome P450 ( CYP ) 2C8 , P11712 REA , CYP 3A , and P10635 REA , as well as P-glycoprotein ( P-gp ) . Based on these results , a series of clinical drug-drug interaction ( DB00900 ) studies were conducted to evaluate the effect of ivacaftor on sensitive substrates of P10632 REA ( rosiglitazone ) , CYP 3A ( midazolam ) , P10635 REA ( desipramine ) , and P-gp ( digoxin ) . In addition , a DB00900 study was conducted to evaluate the effect of ivacaftor on a combined oral contraceptive , as this is considered an important comedication in CF patients . The results indicate ivacaftor is a weak inhibitor of CYP 3A and P-gp , but has no effect on P10632 REA or P10635 REA . DB08820 MENMAX DB08820 MEN caused non-clinically significant increases in ethinyl estradiol and norethisterone exposure . Based on these results , caution and appropriate monitoring are recommended when concomitant substrates of P11712 REA , CYP 3A and / or P-gp are used during treatment with ivacaftor , particularly drugs with a narrow therapeutic index , such as warfarin .

Assessment of partially deoxygenated deoxynojirimycin derivatives as glucosylceramide synthase inhibitors . Q16739 REA ( Q16739 REA ) is an approved drug target for the treatment of Gaucher disease and is considered as a valid target for combating other human pathologies , including type 2 diabetes . The clinical drug N-butyldeoxynojirimycin ( DB00419 MEN ) is thought to inhibit through mimicry of its substrate , ceramide . In this work we demonstrate that , in contrast to what is proposed in this model , the P06681 REA - hydroxyl of the deoxynojirimycin core is important for Q16739 REA inhibition . Here we show that P13671 REA - OH appears of less important , which may set guidelines for the development of Q16739 REA inhibitors that have less affinity ( in comparison with DB00419 MEN ) for other glycoprocessing enzymes , in particular those hydrolases that act on glucosylceramide .

Characterization of gonadotropin-releasing hormone analogs based on a sensitive cellular luciferase reporter gene assay . A novel cellular assay for the functional characterization of agonistic and antagonistic analogs of gonadotropin-releasing hormone ( DB00644 ) was developed . This assay is based on a fusion of the c-fos immediate-early gene promoter to Photinus pyralis luciferase ( Luc ) as a reporter gene , stably transfected in a recombinant cell line expressing the human P30968 REA . Transcription of endogenous c-fos and fos-Luc fusion gene are transiently induced quite similar by fetal calf serum or the superagonistic analog [ D-Trp 6 ] DB00644 in a selected cell line . The reporter gene was therefore used to monitor agonist-induced signaling via the human P30968 REA . Whereas Luc activity was induced in a dose-dependent manner by DB00644 or [ D-Trp 6 ] DB00644 , different antagonistic peptides completely inhibited this stimulation . The antagonistic potency ( IC50 ) of various peptides with DB00050 MEN and Antarelix as lead compounds in general correlated well with the binding affinity ( KD ) as determined from ligand binding experiments . The specificity of an inhibitory effect was confirmed by P30968 REA - independent stimulation with the phorbol ester 12 - O-tetradecanoylphorbol 13 - acetate or basic fibroblast growth factor . Since this new reporter gene assay is sensitive and simple and can be performed in a microtiter plate , it will significantly facilitate screening and functional characterization of DB00644 analogs .

Polymorphisms influencing olanzapine metabolism and adverse effects in healthy subjects . OBJECTIVE : The pharmacokinetics of olanzapine and response to treatment could be affected by polymorphisms in genes coding for drug-metabolizing enzymes , transporters , or receptors . The aim of this study was to identify genetic markers predictive of pharmacokinetics , pharmacodynamics , and adverse effects of olanzapine . METHODS : Sixty-three healthy volunteers receiving a single 5 - mg oral dose of olanzapine were genotyped for 39 genetic variants that could be related to the response to olanzapine . All genetic variants were analyzed by PharmaChip , but P14416 REA Taq 1A polymorphism was determined by real-time polymerase chain reaction . Olanzapine was measured using high-performance liquid chromatography combined with tandem mass spectrometry . The relationship of gender and polymorphisms with olanzapine pharmacokinetics , the change in prolactin levels , and the incidence of adverse effects were evaluated by multiple regression analysis . RESULTS : The pharmacokinetics of olanzapine was influenced by polymorphisms in P20815 REA , P21266 REA , and Q13224 REA . P01236 REA levels were affected by gender and polymorphisms in P14416 REA and 5 - P28223 REA . Polymorphisms in P11712 REA , P51580 REA , P22309 REA , P08183 REA , and 5 - P28223 REA were related to some adverse effects of olanzapine . CONCLUSIONS : Several polymorphisms can explain differences in the pharmacokinetics , pharmacodynamics , and safety of olanzapine in healthy subjects . Whether these genetic factors influence the risk of therapeutic failure or tolerability in patients remains to be established .

Genotoxicity and induction of DNA damage responsive genes by food-borne heterocyclic aromatic amines in human hepatoma HepG 2 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 CYP 1A 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 HepG 2 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 REA , P05177 REA and P22309 REA and expression of P04637 REA and its downstream regulated genes P38936 REA , GADD 45α and Q07812 REA . Consistent with the up-regulation of P38936 REA and GADD 45α 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 REA and down-regulation of Q07812 REA may indicate that HAAs could suppress apoptosis meaning higher probability of damaged cells to survive and mutate .

A meta-analysis of antipseudomonal penicillins and cephalosporins in pediatric patients with fever and neutropenia . BACKGROUND : Antipseudomonal penicillins ( P05067 REA ) and antipseudomonal cephalosporins ( P25054 REA ) play important roles in the management of pediatric patients with fever and neutropenia ( FN ) . Our primary objective was to describe the risk of treatment failure in children treated with an P05067 REA or P25054 REA as initial empiric therapy for FN . Our secondary objectives were to compare P05067 REA with P25054 REA and third - with fourth-generation P25054 REA as initial empiric therapy in this population . METHODS : We performed electronic searches of Ovid Medline , EMBASE , and the Cochrane Central Register of Controlled Trials , limiting studies to prospective pediatric trials in FN in which at least 1 treatment arm consisted of an P05067 REA or P25054 REA antibiotic with or without an aminoglycoside . Data abstraction was conducted by 2 independent reviewers . RESULTS : From 7281 reviewed articles , 41 studies comprising 51 treatment regimens were included in the meta-analysis . Treatment failure , including antibiotic modification , occurred in 34 % and 41 % of patients treated with P05067 REA and P25054 REA monotherapy , respectively , and 41 % and 33 % of patients treated with P05067 REA - and P25054 REA - aminoglycoside combination therapy , respectively . There were no statistically significant differences in treatment failure including modification , mortality , or adverse events when comparing P05067 REA with P25054 REA monotherapy , P05067 REA with P25054 REA combination therapy , or third - with fourth-generation P25054 REA therapy . CONCLUSIONS : Our meta-analysis suggests that P05067 REA and P25054 REA monotherapy , as well as combination therapy with an aminoglycoside , are efficacious and safe therapeutic options for the empiric management of pediatric patients with FN . Specific antibiotic selection should be based on other important factors , such as cost , availability , and local epidemiologic and resistance patterns .

Inhibition of innate co-receptor Q9NP99 signaling reduces P01730 REA ( + ) T cell activation and prolongs cardiac allograft survival . The innate receptor " triggering-receptor-expressed-on-myeloid-cells - 1 " ( Q9NP99 ) enhances downstream signaling of " pattern recognition receptor " ( PRR ) molecules implicated in inflammatory responses . However the mechanistic role of Q9NP99 in chronic heart rejection has yet to be elucidated . We examined the effect of Q9NP99 ( + ) antigen-presenting cells ( P25054 REA ) on alloreactive P01730 REA ( + ) lymphocytes . Bm12 donor hearts were transplanted into wild-type MHC-class-II-mismatched C57BL / 6J recipient mice . Progressive allograft rejection of bm12 - donor hearts with decreased organ function , severe vasculopathy and allograft fibrosis was evident within 4 weeks . Q9NP99 ( + ) CD11b ( + ) MHC-II ( + ) F4 /8 0 ( + ) P41597 REA ( + ) P25054 REA and IFNγ-producing P01730 REA ( + ) cells were detected during chronic rejection . Peptide inhibition of Q9NP99 attenuated graft vasculopathy , reduced graft-infiltrating leukocytes and prolonged allograft survival , while being accompanied by sustained low levels of P01730 REA ( + ) and CD8 ( + ) cell infiltration . Remarkably , temporary inhibition of Q9NP99 during early immune activation was sufficient for long-term allograft survival . Mechanistically , Q9NP99 inhibition leads to reduced differentiation and proliferation of IFNγ-producing Th1 cells . In conclusion , Q9NP99 influences chronic heart rejection by regulating the infiltration and differentiation of P01730 REA ( + ) lymphocytes .

[ Toxicogenetics of antiretroviral treatment ( II ) : neurotoxicity , hepatotoxicity , lactic acidosis , kidney damage , and other adverse effects of antiretroviral drugs ] . Several pharmacogenetics studies have analyzed the influence of specific genetic polymorphisms on the toxicity of antiretroviral treatment . The present review describes some of the adverse effects of antiretroviral drugs in which a genetic predisposition may be involved : efavirenz-induced neurological toxicity , generally associated with the 516G > T polymorphism of liver enzyme cytochrome P450 2B6 ( P20813 REA ) ; hypersensitivity reactions to nevirapine , associated with specific alleles of major histocompatibility complex , mainly the HLA - Q8IUH3 * 0101 allele , which , in combination with a high P01730 REA lymphocyte count , has been associated with systemic reactions and hepatitis in Caucasians , and the HLA-Cw 8 allele , which is associated with hypersensitivity reactions in persons from the Italian island of Sardinia and from Japan ; nevirapine-induced hepatotoxicity associated with the C > T polymorphism in position 3435T of the P08183 REA ( MDR - 1 ) gene codifying for glycoprotein P ( lower risk ) ; hyperbilirubinemia in patients exposed to atazanavir or indinavir carrying the P22309 REA * 28 polymorphism ; peripheral neuropathy with nucleoside analogues associated with haplogroup T of the mitochondrial genome ( higher risk ) and with the Q30201 REA C282Y genotype of the hemochromatosis gene ( lower risk ) ; the mutation in codon 964 ( R964C ) of the P54098 gene that codifies the mitochondrial polymerase DNA gamma described in a Thai patient with lactic acidosis ; the Q92887 REA gene haplotypes associated with tenofovir-induced proximal tubulopathy , and the risk of pancreatitis in persons with mutations in the P13569 REA and SPINK - 1 genes .

Discovery of DB05130 MEN , a Potent , Selective , and Orally Bioavailable hCCR 2 Antagonist . We report the identification of 13 ( DB05130 MEN ) as a potent human P41597 REA ( hCCR 2 ) antagonist . DB05130 MEN exhibited an IC50 of 3.7 nM in antagonism of monocyte chemoattractant protein - 1 binding to hCCR 2 , an IC50 of 4.7 nM in antagonism of chemotaxis activity , an IC50 of 84 μM in inhibition of the hERG potassium current , a free fraction of 58 % in protein binding , high selectivity over other chemokine receptors and G-protein-coupled receptors , and acceptable oral bioavailability in rodents and primates . In human clinical trials , DB05130 MEN exhibited a pharmacokinetic profile suitable for once-a-day dosing ( T 1/2 = 15 h ) .

Lessons learned from the irinotecan metabolic pathway . DB00762 SUB , a camptothecin analogue , is a prodrug which requires bioactivation to form the active metabolite SN - 38 . SN - 38 acts as a P11387 REA poison . DB00762 SUB has been widely used in the treatment of metastatic colorectal cancer , small cell lung cancer and several other solid tumors . However , large inter-patient variability in irinotecan and SN - 38 disposition , as well as severe but unpredictable diarrhea limits the clinical potential of irinotecan . Intense clinical pharmacology studies have been conducted to elucidate its complicated metabolic pathways and to provide scientific rationale in defining strategies to optimize drug therapy . DB00762 SUB is subjected to be shunted between P08684 REA mediated oxidative metabolism to form two inactive metabolites P25054 REA or NPC and tissue carboxylesterase mediated hydrolysis to form SN - 38 which is eventually detoxified via glucuronidation by P22309 REA to form SN - 38G . The pharmacology of this compound is further complicated by the existence of genetic inter-individual differences in activation and deactivation enzymes of irinotecan ( e . g . , P08684 REA , P20815 REA , P22309 REA ) and sharing competitive elimination pathways with many concomitant medications , such as anticonvulsants , St . John ' s Wort , and ketoconazole . Efflux of the parent compound and metabolites out of cells by several drug transporters ( e . g . , Pgp , Q9UNQ0 , MRP 1 , Q92887 REA ) also occurs . This review highlights the latest findings in drug activation , transport mechanisms , glucuronidation , and CYP 3A - mediated drug-drug interactions of irinotecan in order to unlock some of its complicated pharmacology and to provide ideas for relevant future studies into optimization of this promising agent .

DB00996 MEN prevents oxaliplatin-induced central sensitization in the dorsal horn neurons in rats . OBJECTIVES : The present study aims to study the alteration of glutamatergic transmission in the dorsal horn neurons and the effect of gabapentin on oxaliplatin-induced neuropathic pain in rats . MATERIALS AND METHODS : DB00526 ( 5 mg / kg ) or saline was administered to adult male Sprague-Dawley rats . DB00996 MEN ( 60 mg / kg , IP ) or vehicle was injected daily . Mechanical allodynia was assessed using a series of von Frey filaments . The expression of glutamate receptor subunits ( Q13224 REA and GluR 1 ) and brain-derived neurotrophic factor ( P23560 REA ) was measured in the dorsal horn . The glutamatergic strength was recorded in the spinal cord slices . RESULTS : Administration of oxaliplatin induced significant hyperreactivity to mechanical stimuli in rats , which was attenuated by gabapentin . Significant increase in the expression of P23560 REA was found in the dorsal horn in rats receiving oxaliplatin , which was prevented by gabapentin . Further studies also observed a significant increase in the expression of GluR 1 and Q13224 REA , as well as enhanced glutamatergic transmission in the dorsal horn neurons in rats treated with oxaliplatin . The upregulation of glutamatergic transmission was significantly reversed by gabapentin . CONCLUSION : These results illustrated an increased expression of P23560 REA and enhanced glutamatergic transmission in rats with oxaliplatin-induced neuropathic pain , which was markedly attenuated by gabapentin .

Detection of thymidylate synthase modulators by a novel screening assay . P04818 REA ( 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 MEN ) ; 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 REA cells with DB00544 , DB01629 , FUdR , DB00432 MEN , 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 .

[ Quantitative analysis of P11387 REA activity in human and rat glioma : characterization and mechanism of resistance to antitopoisomerase chemical , camptothecin - 11 ] . DB00762 SUB ( CPT - 11 ) is a new derivation of camptothecin , a plant alkaloid antitumor agent . Previous studies indicated that antitumor activity of CPT - 11 was mediated through interaction of the drugs with its target enzyme , P11387 REA ( topo I ) . In this study , we studied the relation between sensitivity to CPT - 11 and topo I activity of glioma cells . Furthermore , we established CPT - 11 resistant cell lines in order to elucidate potential mechanisms of drug resistance . A clear correlation between the sensitivities to CPT - 11 and topo I activities in surgical glioma specimens was demonstrated . Activities of topo I in CPT - 11 sensitive group ( IC50 values for CPT - 11 ; < 50 micrograms / ml ) tended to be higher than those in CPT - 11 resistant group ( IC50 values ; > or = 50 ) . Topo I activity may serve as a novel marker to predict the sensitivity of gliomas to topo inhibitors . CPT - 11 resistance cell lines ( T98G / CPT - 11 and P13671 REA ) respectively exhibit a 5.4- and 7.3- fold increase in resistance to CPT - 11 . No differences in topo I activity and intracellular accumulation of CPT - 11 were observed between parent and CPT - 11 resistant lines . On the other hand , topo I from T98G / CPT - 11 and P13671 REA / CPT - 11 cells were at least 4 - and 2 - fold resistant to the inhibitory effect of the CPT - 11 on the relaxation activity of topo I in comparison with their parent lines . This enzymological difference may be responsible for the resistance to CPT - 11 .

The human gonadotropin releasing hormone type I receptor is a functional intracellular GPCR expressed on the nuclear membrane . The mammalian type I gonadotropin releasing hormone receptor ( P30968 REA ) is a structurally unique G protein-coupled receptor ( GPCR ) that lacks cytoplasmic tail sequences and displays inefficient plasma membrane expression ( PME ) . Compared to its murine counterparts , the primate type I receptor is inefficiently folded and retained in the endoplasmic reticulum ( ER ) leading to a further reduction in PME . The decrease in PME and concomitant increase in intracellular localization of the mammalian DB00644 - RI led us to characterize the spatial distribution of the human and mouse DB00644 receptors in two human cell lines , P29320 REA 293 and HTR - 8/ SVneo . In both human cell lines we found the receptors were expressed in the cytoplasm and were associated with the ER and nuclear membrane . A molecular analysis of the receptor protein sequence led us to identify a putative monopartite nuclear localization sequence ( NLS ) in the first intracellular loop of DB00644 - RI . Surprisingly , however , neither the deletion of the NLS nor the addition of the Xenopus P30968 REA cytoplasmic tail sequences to the human receptor altered its spatial distribution . Finally , we demonstrate that DB00644 treatment of nuclei isolated from P29320 REA 293 cells expressing exogenous DB00644 - RI triggers a significant increase in the acetylation and phosphorylation of histone H3 , thereby revealing that the nuclear-localized receptor is functional . Based on our findings , we conclude that the mammalian DB00644 - RI is an intracellular GPCR that is expressed on the nuclear membrane . This major and novel discovery causes us to reassess the signaling potential of this physiologically and clinically important receptor .

High expression levels of P48061 REA and P61073 REA predict recurrence of adamanti-nomatous craniopharyngiomas in children . BACKGROUND : Adamantinomatous craniopharyngioma ( O14561 REA ) is a benign but maldevelopmental tumor with a high recurrence rate . OBJECTIVE : Theaim of this study was to investigate the dysregulated biological molecules that play important roles in the recurrence of O14561 REA . METHODS : We first performed microarray analysis on tumor samples from two pediatric patients with recurrent O14561 REA and from two pediatric O14561 REA patients without recurrence after a one-year follow-up . The expression of P48061 REA and P61073 REA in 45 specimens of pediatric O14561 REA was further evaluated by immunohistochemistry . These results were correlated with the clinicopathological parameters and survival of the patients . RESULTS : Four downregulated genes ( P25054 REA , ITGA , P43121 , and Q99727 ) and 16 upregulated genes ( O76096 , P43235 REA , P07711 REA , P48061 REA , P61073 REA , P02751 REA , Q96DB9 , P05106 REA , P08253 REA , P08254 REA , P09237 REA , P14780 REA , Q92570 REA , Q03405 REA , P16035 REA , and P15692 REA ) were found in the recurrent patients . P48061 REA and P61073 REA were highly expressed in 13 patients ( 28.9 % ) and 14 patients ( 31.1 % ) , respectively . High levels of P48061 REA and P61073 REA expression were significantly associated with a poor recurrence-free survival and were the prognostic factors for O14561 REA recurrence in pediatric patients . CONCLUSIONS : High levels of P48061 REA and P61073 REA expression were associated with O14561 REA recurrence . The role of P48061 REA and P61073 REA in the development of brain tumors requires further research .

Novel camptothecin analogues that circumvent Q9UNQ0 - associated drug resistance in human tumor cells . DB00762 SUB ( 7 - ethyl - 10 - [ 4 - ( 1 - piperidino ) - 1 - piperidino ] - carbonyloxycamptothecin ; CPT - 11 ) is a widely used potent antitumor drug that inhibits mammalian P11387 REA ( Topo I ) ; however , overexpression of Q9UNQ0 ( Q9UNQ0 / Q9UNQ0 / Q9UNQ0 ) can confer cancer cell resistance to SN - 38 , the active form of CPT - 11 . We have recently demonstrated that plasma membrane vesicles prepared from Q9UNQ0 - overexpressing PC - 6 / Q8WUX1 - 5H cells transported SN - 38 and its glucuronide conjugate in an DB00171 - dependent manner ( Nakatomi et al . , Biochem Biophys Res Commun 2001 ; 288:827- 32 ) . In the present study , we have characterized a total of 14 new camptothecin ( CPT ) analogues with respect to both the inhibition of Topo I and the substrate specificity of Q9UNQ0 . All of the tested CPT analogues , which have different substitutions at positions 10 and 11 , strongly inhibited the Topo I activity in a cell-free system , as did SN - 38 . Their antitumor activities in the SN - 38 - resistant PC - 6 / Q8WUX1 - 5H2 cell line greatly varied , however , being correlated with intracellular accumulation levels . We have examined DB00171 - dependent transport of those CPT analogues by using plasma membrane vesicles prepared from both PC - 6 / Q8WUX1 - 5H2 cells and Q9UNQ0 - transfected P29320 REA - 293 cells . Based on the substrate specificity of Q9UNQ0 thus evaluated , it is strongly suggested that CPT analogues with high polarity are good substrates for Q9UNQ0 and are therefore effectively extruded from cancer cells . In this context , to circumvent Q9UNQ0 - associated drug resistance , low-polarity CPT analogues are considered to be potent lead compounds . The present study provides a practical approach to discover new CPT-based drugs for the chemotherapy of drug-resistant human cancer .