interacts_with DB00642?
DB00642 : mRNA expression of the target genes TS , GARFT and P00374 correlates with the in vitro chemosensitivity of human solid tumors . 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 . Integrative analysis of transcriptomics , proteomics , and metabolomics data of white adipose and liver tissue of high-fat diet and rosiglitazone-treated insulin-resistant mice identified pathway alterations and molecular hubs . The incidences of obesity and type 2 diabetes are rapidly increasing and have evolved into a global epidemic . In this study , we analyzed the molecular effects of high-fat diet ( HFD ) -induced insulin-resistance on mice in two metabolic target tissues , the white adipose tissue ( WAT ) and the liver . Additionally , we analyzed the effects of drug treatment using the specific PPARγ ligand rosiglitazone . We integrated transcriptome , proteome , and metabolome data sets for a combined holistic view of molecular mechanisms in type 2 diabetes . Using network and pathway analyses , we identified hub proteins such as P21912 and P53597 in WAT and deregulation of major metabolic pathways in the insulin-resistant state , including the TCA cycle , oxidative phosphorylation , and branched chain amino acid metabolism . Rosiglitazone treatment resulted mainly in modulation via Q07869 signaling and oxidative phosphorylation in WAT only . Interestingly , in HFD liver , we could observe a decrease of proteins involved in vitamin B metabolism such as Q6P996 and P00374 and the according metabolites . Furthermore , we could identify sphingosine ( Sph ) and sphingosine 1-phosphate ( SP1 ) as a drug-specific marker pair in the liver . In summary , our data indicate physiological plasticity gained by interconnected molecular pathways to counteract metabolic dysregulation due to high calorie intake and drug treatment . The anti-inflammatory drug BAY 11-7082 suppresses the MyD88-dependent signalling network by targeting the ubiquitin system . The compound BAY 11-7082 inhibits IκBα [ inhibitor of NF-κB ( nuclear factor κB ) α ] phosphorylation in cells and has been used to implicate the canonical IKKs ( IκB kinases ) and NF-κB in > 350 publications . In the present study we report that BAY 11-7082 does not inhibit the IKKs , but suppresses their activation in LPS (lipopolysaccharide)-stimulated RAW macrophages and IL (interleukin)-1-stimulated IL-1R ( IL-1 receptor ) P29320 ( human embryonic kidney ) -293 cells . BAY 11-7082 exerts these effects by inactivating the E2-conjugating enzymes Ubc ( ubiquitin conjugating ) 13 and P68036 and the E3 ligase LUBAC ( linear ubiquitin assembly complex ) , thereby preventing the formation of Lys63-linked and linear polyubiquitin chains . BAY 11-7082 prevents ubiquitin conjugation to P61088 and P68036 by forming a covalent adduct with their reactive cysteine residues via Michael addition at the P01024 atom of BAY 11-7082 , followed by the release of 4-methylbenzene-sulfinic acid . BAY 11-7082 stimulated Lys48-linked polyubiquitin chain formation in cells and protected HIF1α ( hypoxia-inducible factor 1α ) from proteasomal degradation , suggesting that it inhibits the proteasome . The results of the present study indicate that the anti-inflammatory effects of BAY 11-7082 , its ability to induce B-cell lymphoma and leukaemic T-cell death and to prevent the recruitment of proteins to sites of DNA damage are exerted via inhibition of components of the ubiquitin system and not by inhibiting NF-κB . P15941 in macrophage : contributions to cigarette smoke-induced lung cancer . Expression of the pro-oncogenic mucin P15941 is elevated by inflammation in airway epithelial cells , but the contributions of P15941 to the development of lung cancer are uncertain . In this study , we developed our finding that cigarette smoke increases Muc1 expression in mouse lung macrophages , where we hypothesized P15941 may contribute to cigarette smoke-induced transformation of bronchial epithelial cells . In human macrophages , cigarette smoke extract ( CSE ) strongly induced P15941 expression through a mechanism involving the nuclear receptor Q07869 -γ . CSE-induced extracellular signal-regulated kinase ( P29323 ) activation was also required for P15941 expression , but it had little effect on P15941 transcription . RNA interference-mediated attenuation of P15941 suppressed CSE-induced secretion of P01375 -α from macrophages , by suppressing the activity of the P01375 -α-converting enzyme ( P78536 ) , arguing that P15941 is required for CSE-induced and P78536 -mediated P01375 -α secretion . Similarly , P15941 blockade after CSE induction through suppression of Q07869 -γ or P29323 inhibited P78536 activity and P01375 -α secretion . Conditioned media from CSE-treated macrophages induced P15941 expression and potentiated CSE-induced transformation of human bronchial epithelial cells in a P01375 -α-dependent manner . Together , our results identify a signaling pathway involving Q07869 -γ , P29323 , and P15941 for P01375 -α secretion induced by CSE from macrophages . Furthermore , our results show how P15941 contributes to smoking-induced lung cancers that are driven by inflammatory signals from macrophages . Regulation of hepatic ApoC3 expression by P20142 -1β mediates hypolipidemic effect of nicotinic acid . Peroxisome proliferator-activated receptor ( Q07869 ) γ coactivator-1β ( P20142 -1β ) is a transcriptional coactivator that induces hypertriglyceridemia in response to dietary fats through activating hepatic lipogenesis and lipoprotein secretion . The expression of P20142 -1β is regulated by free fatty acids . Here we show that P20142 -1β regulates plasma triglyceride metabolism through stimulating apolipoprotein P01024 ( P02656 ) expression and elevating P02656 levels in circulation . Remarkably , liver-specific knockdown of P02656 significantly ameliorates P20142 -1β-induced hypertriglyceridemia in mice . Hepatic expression of P20142 -1β and P02656 is reduced in response to acute and chronic treatments with nicotinic acid , a widely prescribed drug for lowering plasma triglycerides . Adenoviral-mediated knockdown of P20142 -1β or P02656 in the liver recapitulates the hypolipidemic effect of nicotinic acid . Proteomic analysis of hepatic P20142 -1β transcriptional complex indicates that it stimulates P02656 expression through coactivating orphan nuclear receptor ERRα and recruiting chromatin-remodeling cofactors . Together , these studies identify P20142 -1β as an important regulator of the P02656 gene cluster and reveal a mechanism through which nicotinic acid achieves its therapeutic effects . Neuroinflammation in Alzheimer 's disease : chemokines produced by astrocytes and chemokine receptors . Chemokines secreted by astrocytes play multiple roles in the pathology of Alzheimer 's disease , a chronic inflammation disorder of central nervous system . The level of chemokines in serum , cerebrospinal fluid and brain tissue and their receptors both significantly changed in patients with Alzheimer 's disease . In this review , we briefly summarized the involvement of astrocytes and chemokines in Alzheimer 's disease , and the role of chemokine/chemokine receptors in the occurrence and development of Alzheimer 's disease . Clarification of the involvement of chemokines and their receptors , such as P13500 / P41597 , fractalkine/ P49238 , SDF-1α/ P61073 , MIP-1α/ P51681 , P02778 / P49682 , P10145 / P25024 , P25025 , and RANTES/ P32246 , P51677 , P51681 , will provide a new strategy and more specific targets for the treatment of Alzheimer 's disease . IL-1 transcriptionally activates the neutrophil chemotactic factor/ P10145 gene in endothelial cells . Leucocytes and vascular cells interact closely in inflammation and immunity and cytokines are important mediators of this interaction . The present study was designed to define the capacity of human endothelial cells ( O14777 ) to produce a monocyte-derived neutrophil chemotactic factor ( provisionally termed P10145 ) . P10145 is a polypeptide chemotactic for neutrophils originally identified in the culture supernatant of lipopolysaccharide ( LPS ) -stimulated monocytes . IL-1 induced high levels of production of neutrophil chemotactic activity in culture supernatants of O14777 . Optimal stimulation of activity was observed when O14777 were cultured with 10-100 ng/ml P01584 for 16 hr . Anti- P10145 antibody blocked the chemotactic activity for neutrophils of IL-1-activated O14777 supernatants . IL-1-treated O14777 expressed high levels of P10145 mRNA transcripts , as assessed by Northern blot analysis . Tumour necrosis factor ( P01375 ) and LPS , unlike the inflammatory monokine P05231 , also induced P10145 expression . Nuclear run-off experiments revealed that IL-1 activated transcription of the P10145 gene . The production of P10145 may represent a mechanism whereby endothelial cells , exposed to inflammatory signals , participate in the regulation of neutrophil extravasation . Autologous P15941 -specific Th1 effector cell immunotherapy induces differential levels of systemic TReg cell subpopulations that result in increased ovarian cancer patient survival . Adoptive T cell immunotherapy using autologous lymphocytes is a viable treatment for patients with cancer and requires participation of Ag-specific P01730 and CD8 T cells . Here , we assessed the immunotherapeutic effects of autologous P15941 peptide-stimulated P01730 (+) effector cells following adoptive transfer in patients with ovarian cancer . Using P15941 peptide and P60568 for ex vivo P01730 (+)/Th1 effector cell generation , we show that three monthly treatment cycles of peripheral blood T cell restimulation and intraperitoneal re-infusion selectively modulated endogenous T cell-mediated immune responses that correlated with diminished serum Q8WXI7 tumor marker levels and enhanced patient survival . One patient remains disease-free , another patient survived long-term for nearly 16 months with recurrent disease and two patients expired within 3-5 months following final infusion . Although PBL from all patients showed elevated P15941 cytolytic activity following therapy , such responses did not correlate with therapeutic efficacy . Long-term survivors showed elevated levels of systemic memory ( CD45RO ) and naïve ( CD45RA ) CD3/ P01730 /CD25(+) T cells when compared to that of pre-treatment levels and similarly treated short-term survivors . Such cells co-expressed different levels of Foxp3 and P16410 that resulted in progressively lower systemic Foxp3/ P16410 memory T cell ratios that further correlated with disease-free survival . Lastly , these patients showed elevated levels of P15941 -specific T cells expressing the P51681 and P32246 chemokine receptors and the chemokine P13236 associated with Th1 cell differentiation/memory . We suggest that effective immunotherapy with autologous P15941 -stimulated P01730 (+) effector cells induces differential levels of systemic " Ag-experienced " and " Ag-inexperienced " P01730 /CD25(+) TReg cell subpopulations that influence long-term tumor immunity in ovarian cancer patients . Metabolism of risperidone to 9-hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9-hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 , P05177 , P10632 , P11712 -arg144 , P11712 -cys144 , P33261 , P10635 , P08684 and P20815 supplemented with an NADPH-generating system . DB01267 was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9-hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol(-1) CYP min(-1) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9-hydroxyrisperidone is highly correlated with P10635 and 3A activities . Thus , both P10635 and 3A4 are involved in the 9-hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 ) and ketoconazole ( inhibitor of P08684 ) can inhibit the formation of 9-hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9-hydroxyrisperidone in rat . The formation of 9-hydroxyrisperidone is highly correlated with testosterone 6beta-hydroxylase activities , suggesting that inducible CYP3A contributes significantly to the metabolism of risperidone in rat . New data integrating multitargeted antifolates into treatment of first-line and relapsed non-small-cell lung cancer . Non-small-cell lung cancer ( NSCLC ) represents approximately 80 % of all lung cancers . With modern platinum- based combination regimens , overall median survival has reached 9-12 months . Antifolates are active against several solid tumors and hematologic malignancies . The cytotoxic action of antifolates is mainly related to their ability to inhibit several different folate-dependent enzymes involved in DNA synthesis . DB00642 is a novel multitargeted antifolate that inhibits at least 3 of the enzymes involved in purine and pyrimidine synthesis : thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyltransferase ( GARFT ) . DB00642 was approved for the treatment of relapsed NSCLC as it produced equivalent response and survival rates and less toxicity compared with docetaxel . DB00642 in combination with platinum analogues or with gemcitabine showed equivalent clinical impact compared with standard combinations of platinum plus third-generation agents . We analyze the potential implications of pemetrexed 's role in first-line chemotherapy of NSCLC as well as hints of differential cytotoxic action according to histology , new schedules of vitamin supplementation , and target enzymes expression levels . Issues of pharmacogenomics are becoming relevant in defining pemetrexed efficacy . Chemosensitivity was significantly linked to low levels of TS , GARFT , and P00374 in preclinical models . Consequently , the differential expression of TS according to histology might explain the different activity of pemetrexed according to histology , as recently postulated . Development of stable cell lines expressing different subtypes of GABAA receptors . The experiments reported here were motivated by our interest to express in stably-transfected cells large amounts of recombinant rat GABAA receptors . For this , we developed an original two step selection strategy , in which the first step consisted of transfecting P29320 293 cells with rat GABAA receptor alpha and beta subunits . G 418 resistant colonies isolated at this step were screened for [ 3H ] muscimol binding to select for those that coexpressed alpha- and beta-subunits . The best alpha and beta subunit expressing colony was then supertransfected with a plasmid coding for the gamma rat GABAA receptor subunit and a mutant P00374 gene . After a second round of selection , this time in presence of methotrexate , those colonies that coexpressed ternary alpha beta gamma GABAA receptor combinations were distinguished using [ 3H ] flumazenil as a probe . This strategy was applied to the isolation of 3 GABAA receptor clones , alpha 1 beta 2 gamma 2s , alpha 3 beta 2 gamma 2s and alpha 5 beta 3 gamma 2s , that expressed relatively high levels of these proteins . These 3 cell lines exhibited pharmacological and functional properties similar to cells transiently-transfected with equivalent subunit combinations . These cell lines therefore provide attractive models with which to evaluate the intrinsic activity and potency of compounds at recombinant GABAA receptor subtypes . Recent advances in classical and non-classical antifolates as antitumor and antiopportunistic infection agents : Part II . Antifolates that inhibit the key enzymes thymidylate synthase ( TS ) and dihydrofolate reductase ( P00374 ) have found clinical utility as antitumor and antiopportunistic agents . DB00563 { MTX , ( 1 ) } and 5-fluorouracil ( DB00544 ) were among the first clinically useful P00374 and TS inhibitors , respectively . The development of resistance to DB00544 , its occasional unpredictable activity and toxicity resulted in the search of novel antifolates . DB00642 ( 4 ) and raltitrexed ( 5 ) are newer antifolates that specifically inhibit TS , and are clinically useful as antitumor agents . A major mechanism of tumor resistance to clinically useful antifolates is based on their need for polyglutamylation via the enzyme folylpoly-gamma-glutamate synthetase ( Q05932 ) . Recently , classical antifolates that do not need to be polyglutamylated have also been developed and include plevitrexed ( 6 ) and GW1843 ( 7 ) . Nolatrexed ( 8 ) , trimethoprim { P54849 , ( 11 ) } and piritrexim { PTX , ( 12 ) } are nonclassical antifolates for antitumor and parasitic chemotherapy that passively diffuse into cells and hence do not have to depend on Q05932 or the reduced folate carrier ( P41440 ) . Structural requirements for inhibition with antifolates have been studied extensively and novel agents that exploit key interactions in the active site of TS , P00374 , Q05932 , and P41440 have been proposed . This two-part review discusses the design , synthesis and structural requirements for TS and P00374 inhibition and their relevance to antitumor and parasitic chemotherapy , since 1996 . Monocyclic and 6-5 fused bicyclic antifolates were discussed in Part I . The 6-6 bicyclic and tricyclic antifolates will be discussed here in Part II . Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 , Q16678 , P11712 , P33261 , P05181 , P05093 , P11511 , P35869 , P03372 , Q92731 , ERRRG , P06401 , P07099 , P34913 , P37059 , P37058 , P28161 , P21266 , GSTT2 , P09211 , NAT1 , NAT2 , P21964 , P07327 , P00325 , P00326 , P05091 , P35228 , NOS3 , P01583 , P01584 , O15527 , P36639 [ P36639 ] , P14416 , P35462 , P21917 , P31645 , P04150 [ GCCR ] , P42898 , and P15559 . In the present study , the Japanese allele frequencies were verified by using nationwide population samples . DB00642 alters folate phenotype and inflammatory profile in EA.hy 926 cells grown under low-folate conditions . Elevated homocysteine is a risk marker for several major human pathologies . Emerging evidence suggests that perturbations of folate/homocysteine metabolism can directly modify production of inflammatory mediators . DB00642 acts by inhibiting thymidylate synthetase ( P04818 ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyltransferase ( GARFT ) . EA.hy 926 cells grown under low ( " Lo " ) and high ( " Hi " ) folate conditions were treated with pemetrexed . The concentrations of several intracellular folate derivatives were measured using LC-MRM/MS . Lo cells had lower total folate concentrations and a different distribution of the intracellular folate derivatives than Hi cells . Treatment with pemetrexed caused a decrease in individual folate analytes . Microarray analysis showed that several genes were significantly up or down-regulated in pemetrexed treated Lo cells . Several of the significantly up-regulated transcripts were inflammatory . Changes in transcript levels of selected targets , including P01024 , P10145 , and P00374 , were confirmed by quantitative RT-PCR . P01024 and P10145 transcript levels were increased in pemetrexed-treated Lo cells relative to Lo controls ; P00374 transcript levels were decreased . In Lo cells , P10145 and P01024 protein concentrations were increased following pemetrexed treatment . DB00642 drug treatment was shown in this study to have effects that lead to an increase in pro-inflammatory mediators in Lo cells . No such changes were observed in Hi cells , suggesting that pemetrexed could not modify the inflammatory profile in the context of cellular folate sufficiency . Effects of retroviral-mediated P08183 expression on hematopoietic stem cell self-renewal and differentiation in culture . Ex vivo expansion of hematopoietic stem cells would be useful for bone marrow transplantation and gene therapy applications . Toward this goal , we have investigated whether retrovirally-transduced murine stem cells could be expanded in culture with hematopoietic cytokines . Bone marrow cells were transduced with retroviral vectors expressing either the human multidrug resistance 1 gene ( HaMDR1 ) , a variant of human dihydrofolate reductase ( HaDHFR ) , or both P08183 and P00374 in an internal ribosomal entry site ( IRES ) -containing bicistronic vector ( HaMID ) . Cells were then expanded for 15 days in cultures stimulated with interleukin ( IL ) -3 , P05231 , and stem cell factor . When very low marrow volumes were injected into lethally irradiated recipient mice , long-term reconstitution with 100 % donor cells was seen in all mice injected with HaMDR1- or HaMID-transduced cells . By contrast , engraftment with HaDHFR- or mock-transduced cells ranged from partial to undetectable despite injection of significantly larger marrow volumes . In addition , mice transplanted with expanded HaMDR1- or HaMID-transduced stem cells developed a myeloproliferative disorder that was characterized by an increase in abnormal peripheral blood leukocytes . These results show that P08183 -transduced stem cells can be expanded in vitro with hematopoietic cytokines , but indicate that an increased stem cell division frequency can lead to stem cell damage . Establishment of pemetrexed-resistant non-small cell lung cancer cell lines . DB00642 ( P15941 ) , a multitargeted antifolate with manageable toxicity , is active against non-squamous non-small cell lung cancer ; however , most patients eventually acquire resistance to P15941 . To elucidate the resistant mechanism , we established P15941 -resistant lung adenocarcinoma cell lines . Two parental cell lines , PC-9 and A549 , were treated with step-wise increasing concentrations of P15941 . Growth inhibition was determined by the 3-[4,5-dimethyl-thizol-2-yl]-2,5-diphenyltetrazolium bromide assay . Expression of the genes encoding thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyltransferase ( GARFT ) was analyzed by quantitative real-time reverse transcriptase polymerase chain reaction . The four PC-9 sublines were more resistant than the PC-9 cell line to P15941 ( 2.2- , 2.9- , 8.4- , and 14.3-fold , respectively ) . The four A549 sublines also showed more resistance to P15941 ( 7.8- , 9.6- , 42.3- , and 42.4-fold , respectively ) than the parent cell line . All resistant sublines showed cross-resistance to cisplatin , but not to docetaxel , vinorelbine , 5-fluorouracil , or the active metabolite of irinotecan , SN-38 . All P15941 -resistant sublines expressed more TS than the parental cells , by polymerase chain reaction and Western blotting . P00374 was significantly increased in the four P15941 -resistant A549 sublines . GARFT did not correlate with resistance to P15941 . In summary , P15941 -resistant cells remained sensitive to docetaxel , vinorelbine , 5-fluorouracil , and irinotecan . TS expression appeared to be associated with resistance to P15941 . Expression patterns of the implantation-associated genes in the uterus during the estrous cycle in mice . The mRNA expression patterns of P01133 , HB- P01133 , P15514 , P01133 receptor , DB01277 , P09603 , P01583 , P01584 , IL-1 receptor type 1 , IL-1 receptor antagonist , P15018 , P23219 , P35354 , P15941 , calcitonin , and rat Q6X4U4 mouse homologue , all of which are involved in the process of conceptus implantation to the endometrium , were examined during the estrous cycle by means of real-time quantitative PCR . P35354 , HB- P01133 , P15018 , P15941 , P09603 , P01583 , P01584 , and IL-1 receptor antagonist were temporally regulated during the estrous cycle and highly expressed during the estrous stage . In the case of P23219 , P01133 , DB01277 , and P01133 receptor , the highest mRNA expression was during the diestrous stage . In contrast , the rat Q6X4U4 mouse homologue mRNA expression did not change during the estrous cycle . These results indicate that rat Q6X4U4 mouse homologue expression at implantation might be specifically regulated by embryonic factors rather than the maternal environment . DB00945 antagonizes the cytotoxic effect of methotrexate in lung cancer cells . DB00563 ( MTX ) has been widely used for the treatment of cancer and rheumatoid arthritis ( RA ) . DB00945 ( ASA ) is a non-selective cyclooxygenase ( P36551 ) inhibitor that contributes to the treatment of inflammatory conditions such as RA . It has been observed that the antitumor effect of ASA can be attributed to inhibition of cell cycle progression , induction of apoptosis and inhibition of angiogenesis . In the present study , we revealed that the treatment with a combination of MTX and ASA resulted in antagonism of the cytotoxic effect as demonstrated by P50991 and colony formation assays . ASA alleviated the MTX-mediated S phase accumulation and recovered the P55008 phase . MTX-mediated accumulation of the S phase marker cyclin A was also alleviated by ASA . Notably , FAS protein levels were upregulated by MTX in A549 cells . The antagonism of MTX efficacy caused by ASA was accompanied by altered expression of caspase-3 , Bcl-2 and FAS but not dihydrofolate reductase ( P00374 ) . This suggests that the alteration of caspase-3 , Bcl-2 and FAS was involved in the antagonism between ASA and MTX . Exogenously added folic acid reversed the MTX-mediated P00374 inhibition following either MTX or MTX + ASA treatments . Most importantly , we demonstrated for the first time that the commonly used non-steroidal anti-inflammatory drug for headache ASA and possibly other P23219 /2 inhibitors can produce a strong antagonistic effect on the growth inhibition of lung cancer cells when administered in combination with MTX . The clinical implication of our finding is obvious , i.e. , the clinical efficacy of MTX therapy can be compromised by ASA and their concomitant use should be avoided . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . DB00067 triggers senescence in K-ras transformed cells via RhoA-dependent downregulation of cyclin D1 . DB00067 ( AVP ) , a vasoactive peptide hormone that binds to three G-protein coupled receptors ( V1R , P30518 , and V3R ) , has long been known to activate V1R and elicit mitogenesis in several cell types , including adrenal glomerulosa cells . However , in the mouse Q03519 adrenocortical malignant cell line , AVP triggers not only a canonical mitogenic response but also novel RhoA-GTP-dependent mechanisms which downregulate cyclin D1 , irreversibly inhibiting K-ras oncogene-driven proliferation . In Q03519 cells , AVP blocks cyclin D1 expression , induces senescence-associated beta-galactosidase ( SAbeta-Gal ) and inhibits proliferation . However , ectopic expression of cyclin D1 renders Q03519 cells resistant to both SAbeta-Gal induction and proliferation inhibition by AVP . In addition , ectopic expression of the dominant negative RhoAN19 mutant blocks RhoA activation , yielding Q03519 cell sub-lines which are no longer susceptible to cyclin D1 downregulation , SAbeta-Gal induction , or proliferation inhibition by AVP . Furthermore , inhibiting RhoA with P01024 exoenzyme protects Q03519 cells from AVP proliferation inhibition and SAbeta-Gal induction . On the other hand , AVP treatment does not activate caspases 3 and 7 , and the caspase inhibitor Ac-DEVD-CMK does not protect Q03519 cells from proliferation inhibition by AVP , implying that AVP does not trigger apoptosis . These results underline a pivotal survival activity of cyclin D1 that protects K-ras oncogene-dependent malignant cells from senescence . Activation of the MyD88 signaling pathway inhibits ischemia-reperfusion injury in the small intestine . Toll-like receptors ( TLRs ) recognize microbial components and trigger the signaling cascade that activates innate and adaptive immunity . Recent studies have shown that the activation of TLR-dependent signaling pathways plays important roles in the pathogenesis of ischemia-reperfusion ( I/R ) injuries in many organs . All TLRs , except O15455 , use a common adaptor protein , MyD88 , to transduce activation signals . We investigated the role of MyD88 in I/R injury of the small intestine . MyD88 and cyclooxygenase-2 ( P35354 ) knockout and wild-type mice were subjected to intestinal I/R injury . I/R-induced small intestinal injury was characterized by infiltration of inflammatory cells , disruption of the mucosal epithelium , destruction of villi , and increases in myeloperoxidase activity and mRNA levels of P01375 -α and the P10145 homolog KC . MyD88 deficiency worsened the severity of I/R injury , as assessed using the histological grading system , measuring luminal contents of hemoglobin ( a marker of intestinal bleeding ) , and counting apoptotic epithelial cells , while it inhibited the increase in mRNA expression of P01375 -α and KC . I/R significantly enhanced P35354 expression and increased PGE(2) concentration in the small intestine of wild-type mice , which were markedly inhibited by MyD88 deficiency . P35354 knockout mice were also highly susceptible to intestinal I/R injury . Exogenous PGE(2) reduced the severity of injury in both MyD88 and P35354 knockout mice to the level of wild-type mice . These findings suggest that the MyD88 signaling pathway may inhibit I/R injury in the small intestine by inducing P35354 expression . Association of P35462 and Q13224 with impulse control and related behaviors in Parkinson 's disease . We aimed to assess whether allelic variants of dopamine receptor , glutamate receptor , and serotonin transporter genes are associated with the appearance of impulse control and related behaviors ( ICRB ) in Parkinson 's disease ( PD ) with dopamine replacement therapy ( P29323 ) . We surveyed ICRB in consecutive Korean patients with PD who were treated with stable P29323 using modified Minnesota Impulsive Disorders Interview over a period of 4 months . In the 404 patients who completed the interview and the 559 Korean healthy normal controls , genotyping was performed for variants of the P35462 p.S9G , P14416 Taq1A , Q13224 c.366C > G , c.2664C > T and c.-200T > G , and the promoter region of the serotonin transporter gene ( 5-HTTLPR ) . Behavioral abnormalities suggestive of ICRB including compulsive buying , gambling , sexual behavior and eating , and punding , were present in 14.4 % of the patients . Variants of P14416 and 5-HTTLPR were not associated with the risk of developing ICRB . However , the AA genotype of P35462 p.S9G and the CC genotype of Q13224 c.366C > G were more frequent in patients with ICRB than in nonaffected patients ( odds ratio [ OR ] = 2.21 , P = 0.0094 ; and 2.14 , P = 0.0087 , after adjusting for age and sex ) . After controlling for clinical variables in the multivariate analysis , carriage of either AA genotype of P35462 or CC genotype of Q13224 was identified as an independent risk factor for ICRB ( adjusted OR : 2.57 , P = 0.0087 ) . Variants of P35462 p.S9G and Q13224 c.366C > G may be associated with the appearance of ICRB in PD . Cbl-b is a negative regulator of inflammatory cytokines produced by IgE-activated mast cells . c-Cbl and Cbl-b E3 ubiquitin ligases are abundantly expressed in hemopoietic cells where they negatively regulate the activity and levels of many cell surface receptors and associated signaling molecules . By comparing bone marrow-derived mast cells from c-Cbl and Cbl-b-deficient mice it has recently been shown that Cbl-b is the dominant family member for negatively regulating signaling responses from high-affinity IgE receptors . In this study , we suggest that a possible reason for the greater enhancement of IgE receptor signaling in Cbl-b-deficient mice is the relatively higher levels of Cbl-b protein over c-Cbl in mast cells compared with other hemopoietic cells . We also directly compare mast cells from c-Cbl and Cbl-b-deficient mice and find that loss of Cbl-b , but not c-Cbl , increases cell growth , retards receptor internalization , and causes the sustained tyrosine phosphorylation of Syk and its substrates . However , loss of Cbl-b does not enhance the activation of P29323 or Akt , nor does it promote a greater calcium response . Furthermore , loss of Cbl-b or c-Cbl does not increase levels of the Syk or Lyn protein tyrosine kinases . Most notable , however , is the extremely large increase in the production of proinflammatory cytokines P01375 , P05231 , and P13500 by Cbl-b(-/-) mast cells compared with levels produced by c-Cbl(-/-) or wild-type cells . This marked induction , which appears to be restricted to these three cytokines , is dependent on IgE receptor activation and correlates with enhanced O15111 phosphorylation . Thus , Cbl-b functions as a potent negative regulator of cytokines that promote allergic and inflammatory reactions . Celecoxib with chemotherapy in colorectal cancer . P35354 ( P35354 ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 inhibitors , such as sulindac ( DB00605 ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U.S . Food and Drug Administration has approved specific P35354 inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 inhibitors in both prevention and treatment of a diverse range of human cancers . In vivo protection of activated Tyr22-dihydrofolate reductase gene-modified canine T lymphocytes from methotrexate . BACKGROUND : Nonmyeloablative allogeneic hematopoietic stem cell ( P19526 ) transplantation can cure malignant and nonmalignant diseases affecting the hematopoietic system , such as severe combined immunodeficiencies , aplastic anemia and hemoglobinopathies . Although nonmyeloablative is favored over myeloablative transplantation for many patients , graft rejection remains problematic . One strategy for decreasing rejection is to protect donor activated T cells in the graft from methotrexate ( MTX ) by genetically modifying the cells to express MTX-resistant dihydrofolate reductase ( Tyr22- P00374 ) , leaving the immunosuppressive effects of MTX to act solely on activated host T lymphocytes , shifting the balance to favor allogeneic engraftment . METHODS : To evaluate MTX resistance of Tyr22- P00374 (+) T lymphocytes in vivo , we transplanted dogs with autologous P28906 (+) cells modified with yellow fluorescent protein ( YFP ) and P00374 -green fluorescent protein ( GFP ) lentivirus vectors . Dogs were then treated with a standard MTX regimen days 1 , 3 , 6 and 11 ) following immune activation with a foreign antigen as a surrogate assay to mimic early transplantation . RESULTS : P00374 -GFP(+) gene marking was maintained in CD3(+) CD25(+) and P01730 (+) T lymphocytes after MTX treatment , whereas the level of T lymphocytes that expressed only a fluorescent reporter ( YFP(+) ) decreased . These data show that Tyr22- P00374 expression protects T lymphocytes from MTX toxicity in dogs , highlighting a clinically relevant application for preserving donor T lymphocytes during post-transplantation immunosuppression . CONCLUSIONS : The findings of the present study have implications for the clinical translation of MTX-resistant T cells to facilitate engraftment of allogeneic cells following nonmyeloablative conditioning and to minimize the risk of rejection . In summary , Tyr22- P00374 expression in T lymphocytes provides chemoprotection from MTX-mediated elimination in the context of immune activation in vivo . Cellular mechanisms of the hemostatic effects of desmopressin ( DB00035 ) . The synthetic analog of vasopressin desmopressin ( DB00035 ) is widely used for the treatment of patients with von Willebrand disease ( VWD ) , hemophilia A , several platelet disorders , and uremic bleeding . DB00035 induces an increase in plasma levels of P04275 ( P04275 ) , coagulation factor VIII ( FVIII ) , and tissue plasminogen activator ( t-PA ) . It also has a vasodilatory action . In spite of its extensive clinical use , its cellular mechanism of action remains incompletely understood . Its effect on P04275 and t-PA as well as its vasodilatory effect are likely explained by a direct action on the endothelium , via activation of endothelial vasopressin P30518 receptor and DB02527 -mediated signaling . This leads to exocytosis from Weibel Palade bodies where both P04275 and t-PA are stored , as well as to nitric oxide ( NO ) production via activation of endothelial NO synthase . The mechanism of action of DB00035 on FVIII plasma levels remains to be elucidated . The hemostatic effect of DB00035 likely involves additional cellular effects that remain to be discovered . Design , synthesis , and X-ray crystal structures of 2,4-diaminofuro[2,3-d]pyrimidines as multireceptor tyrosine kinase and dihydrofolate reductase inhibitors . To optimize dual receptor tyrosine kinase ( RTK ) and dihydrofolate reductase ( P00374 ) inhibition , the E- and Z-isomers of 5-[2-(2-methoxyphenyl)prop-1-en-1-yl]furo[2,3-d]pyrimidine-2,4-diamines ( 1a and 1b ) were separated by HPLC and the X-ray crystal structures ( 2.0 and 1.4A , respectively ) with mouse P00374 and NADPH as well as 1b with human P00374 ( 1.5A ) were determined . The E- and Z-isomers adopt different binding modes when bound to mouse P00374 . A series of 2,4-diaminofuro[2,3-d]pyrimidines 2-13 were designed and synthesized using the X-ray crystal structures of 1a and 1b with P00374 to increase their P00374 inhibitory activity . Wittig reactions of appropriate 2-methoxyphenyl ketones with 2,4-diamino-6-chloromethyl furo[2,3-d]pyrimidine afforded the Q99618 - P02748 unsaturated compounds 2-7 and catalytic reduction gave the saturated 8-13 . Homologation of the P02748 -methyl analog maintains P00374 inhibitory activity . In addition , inhibition of P00533 and P09619 were discovered for saturated P02748 -homologated analogs 9 and 10 that were absent in the saturated P02748 -methyl analogs . Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells . BACKGROUND : Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers . To determine the plausibility that oral keratinocytes are primary targets of HIV-1 , we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner . RESULTS : To study the fate of HIV-1 , immortalized oral keratinocytes ( OKF6/ O14746 -2 ; O14746 -2 cells ) were characterized for the fate of HIV-specific RNA and DNA . At 6 h post inoculation with X4 or R5-tropic HIV-1 , HIV-1gag RNA was detected maximally within O14746 -2 cells . Reverse transcriptase activity in O14746 -2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP . DB00495 inhibited EGFP expression in a dose-dependent manner , suggesting that viral replication can be supported if receptors are bypassed . Within 3 h post inoculation , integrated HIV-1 DNA was detected in O14746 -2 cell nuclei and persisted after subculture . Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation , suggesting that HIV replication may abort and that infection is non-productive . Within 48 h post inoculation , however , virus harbored by P01730 negative O14746 -2 cells trans infected co-cultured peripheral blood mononuclear cells ( PBMCs ) or MOLT4 cells ( P01730 + P51681 + ) by direct cell-to-cell transfer or by releasing low levels of infectious virions . Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner . CONCLUSION : Oral keratinocytes appear , therefore , to support stable non-replicative integration , while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h . Effect of dietary NaCl on tyrosine hydroxylase in the superior cervical ganglia of Dahl rats . To investigate the involvement of peripheral catecholamines in the development of Dahl-Iwai salt-sensitive ( Q8IX12 /Eis ) hypertension , we performed immunohistochemical staining of tyrosine hydroxylase ( TH ) in the superior cervical ganglia ( SCG ) of Q8IX12 /Eis rats and Dahl-Iwai salt-resistant ( P30518 /Eis ) rats , and in situ hybridization histochemistry for demonstration of TH mRNA localization in the SCG of these rats . Q8IX12 /Eis and P30518 /Eis rats were fed on a high ( 8 % ) salt diet or on a low ( 0.3 % ) salt diet for 4 weeks . Nerve cells in the SCG of Q8IX12 /Eis high salt rats exhibited more intense TH-immunoreactivity ( P < 0.01 ) and hybridization signals ( P < 0.01 ) than those of the other experimental groups . These findings suggest that activation of peripheral sympathetic nerves may account for hypertension in Q8IX12 /Eis rats on a high salt diet . Production of recombinant proteins in Chinese hamster ovary cells overexpressing the subtilisin-like proprotein converting enzyme furin . The proprotein processing enzyme furin is the mammalian prototype of a novel family of subtilisin-like serine endoproteases which possess cleavage specificity for sites involving multiple basic amino acid residues and are involved in the processing of precursor proteins of a variety of regulatory peptides and proteins . One of the limiting steps in the engineering of mammalian cells designed for the overproduction of secreted proteins is the endoproteolytic cleavage of the precursor molecule to its mature biologically active form . The extremely low level of endogenous furin is likely the reason why cells are not able to fully mature overexpressed precursor proteins to their mature form . Here , we report a CHO-derived cell line genetically engineered for the production of high levels of recombinant proteins that need such endoproteolytic maturation . First , the human furin cDNA under the control of the cytomegalovirus early promoter and enhancer was introduced and overexpressed in a P00374 -deficient CHO cell line . A permanent cell line CHO-D3- P09958 was established that expressed biologically active furin . Subsequently , to demonstrate the capacity of CHO-D3- P09958 cells to produce recombinant proteins in a fully matured form , two derivative cell lines were established that overexpressed the P04275 ( P04275 ) and transforming growth factor beta 1 ( TGF beta 1 ) ; CHO-D3- P04275 and CHO-D3-TGF beta 1 , respectively . Both derivative cell lines were able to produce relatively high levels of recombinant protein in a fully matured and biologically active form . Our results illustrate the potential of the CHO-D3- P09958 cell line in the production of recombinant secretory proteins that need endoproteolytic activation at the consensus furin cleavage sequence DB00125 -X-Lys/ DB00125 - DB00125 . Ductal adenocarcinoma of the pancreas usually retained Q13485 and p53 protein status as well as expression of epithelial-to-mesenchymal transition markers and cell cycle regulators at the stage of liver metastasis . There are limited data on the biology of metastatic pancreatic ductal adenocarcinoma ( PDAC ) . The aim of the present study was to compare the expression of immunohistochemical markers that may be involved in the development of metastatic disease in primary PDAC and in synchronous liver metastatic tissues . Thirty-two stains ( corresponding to proteins encoded by 31 genes : Q13485 , P04637 , P62736 , CDH1 , P38936 , O95832 , O14493 , O95471 , P35222 , P00533 , P04626 , P02751 , P08727 , P28482 / P27361 , Q16539 , P46013 , P08253 , P14780 , P15941 ( 3 antibodies ) , P98088 , Q6W4X9 , P42345 , MYC , P48681 , P35354 , P62753 , P23443 , P01137 , P36897 , P08670 ) were evaluated using tissue microarray of 26 pairs of primary PDACs and their liver metastases . There were no significant differences in expression levels of examined proteins between primary and secondary lesions . In particular , metastatic PDAC retained the primary tumour 's Q13485 protein status in all and p53 protein status in all but one case . This surprising homogeneity also involved expression levels of markers of epithelial-to-mesenchymal transition as well as cell cycle regulators studied . In conclusion , the biological profiles of primary PDACs and their liver metastases seemed to be similar . Molecular alterations of PDAC related to a set of immunohistochemical markers examined in the present study were already present at the stage of localized disease . DB00909 block of cloned human T-type voltage-gated calcium channels . DB00909 ( ZNS ) is a multi-target antiepileptic drug reported to be efficient in the treatment of both partial and generalized seizures , with T-type Ca(2+) channel blockade being one of its proposed mechanisms of action . In this study , we systematically investigated electrophysiological effects of ZNS on cloned human Ca(v)3.1-3.3 Ca(2+) channels in a heterologous P29320 -293 expression system using whole cell patch-clamp technique . Concentration-response studies were performed in the range from 5 microM to 2mM for Ca(v)3.2 Ca(2+) channels exhibiting a 15.4-30.8 % reduction of Ca(2+) influx within the maximum therapeutic plasma range ( 50-200 microM ZNS ) . The other T-type Ca(2+) channel entities , Ca(v)3.1 and Q9P0X4 , were even less sensitive to ZNS . Both voltage- and concentration-dependence of inactivation kinetics remained unchanged for Ca(v)3.2 VGCC , whereas Ca(v)3.1 and Q9P0X4 exhibited minor , though significant reduction of inactivation-tau . Interestingly , ZNS block of Ca(v)3.2 VGCCs was not use-dependent and remained unaffected by changes in the holding potential . Steady-state inactivation studies did not display a significant shift in steady-state availability of Ca(v)3.2 channels at 100 microM ZNS ( DeltaV(1/2)=3.1mV , p=0.071 ) . Our studies indicate that ZNS is a moderate blocker of human Ca(v)3 T-type Ca(2+) channels with little or no effect on Ca(v)3.2 Ca(2+) channel inactivation kinetics , use- and state-dependence of blockade . These results suggest that T-type Ca(2+) channel inhibition only partially contributes to the anti-absence activity of ZNS antiepileptic drug . Differential role for phospholipase D1 and phospholipase D2 in 12-O-tetradecanoyl-13-phorbol acetate-stimulated MAPK activation , Cox-2 and P10145 expression . Phospholipase D ( PLD ) is expressed in many tissues and stimulated by growth factors and cytokines . However , the role of PLD in signal transduction is still not well-understood . Human embryonic kidney ( P29320 -293 ) cells exhibit low levels of both Q13393 and O14939 mRNA , however , only Q13393 protein was detected by Western blot . When either isoform of PLD was stably expressed in P29320 -293 cells , we observed an increased PLD activity in a cell-free system and a 12-O-tetradecanoyl-13-phorbol acetate ( TPA ) -stimulated increase in PLD activity in intact cells . This system was then used to elucidate the effects of PLD activity on TPA-stimulated signaling pathways . Two such pathways , the mitogen-activated protein kinases ( MAPK ) , extracellular regulated protein kinase ( P29323 ) and p38 are activated by growth factors and cellular stress , respectively . We found that TPA stimulated P29323 phosphorylation regardless of the expression status of PLD . In contrast to P29323 kinase , P29320 -293 cells were unable to induce p38 phosphorylation by TPA stimulation . When P29320 -293 cells expressed either Q13393 or O14939 , we observed elevated p38 phosphorylation in response to TPA stimulation . The P29323 and p38 MAPKs can also stimulate the expression of both cyclooxygenase-2 ( Cox-2 ) and interleukin-8 ( P10145 ) . We used this system to differentiate the effect of Q13393 or O14939 activity on the expression of Cox-2 and P10145 . Increased Cox-2 and P10145 expression was found only in P29320 -293 cells expressing Q13393 . These data identify a novel role for the Q13393 isoform in the induction of gene expression and provide new insight into the differential role of Q13393 and O14939 in cells . Functional polymorphisms of folate-metabolizing enzymes in relation to homocysteine concentrations in systemic lupus erythematosus . OBJECTIVE : To determine if functional polymorphisms of folate/homocysteine pathway enzymes are associated with homocysteine concentrations and/or coronary artery calcification ( CAC ) scores in patients with systemic lupus erythematosus ( SLE ) and controls . METHODS : We investigated 163 SLE patients and 160 controls . Functional polymorphisms in 6 genes in the folate/homocysteine pathway were genotyped : 5,10-methylenetetrahydrofolate reductase ( P42898 ) 677C > T , P42898 1298A > C , cystathionine ss-synthase ( P35520 ) 844ins68 , methionine synthase ( Q99707 ) 2756A > G , methionine synthase reductase ( Q9UBK8 ) 66A > G , thymidylate synthase ( P04818 ) 1494del6 , and dihydrofolate reductase ( P00374 ) c.86+60_78 . RESULTS : Homocysteine levels were higher in African American SLE patients than Caucasian patients and African American controls . Genotype distributions were significantly different in African American and Caucasian controls for 6 of the 7 polymorphisms . Genotype distributions for each polymorphism did not differ significantly between SLE patients and controls even after stratification by race . Glomerular filtration rate was strongly negatively correlated to homocysteine levels , and was therefore adjusted for as a covariate in the models of the effects of the polymorphisms on homocysteine levels . In SLE patients none of the 7 polymorphisms was associated with homocysteine concentrations . In Caucasian controls only P42898 677C > T and 1298A > C showed effects on homocysteine similar to what would be expected from the literature . There were no genotypic associations with median CAC scores in SLE patients or controls with and without stratification by race . CONCLUSION : Polymorphisms in folate/homocysteine metabolizing enzymes do not predict higher homocysteine levels or CAC scores in patients with SLE . P15941 expression is repressed by protein inhibitor of activated signal transducer and activator of transcription-y . Mucin 1 ( P15941 ) is a transmembrane glycoprotein that modulates the interaction between the embryo and the uterine epithelial cell surface . P15941 also is a tumor marker and has been implicated in the protection of cancer cells from immune cell attack as well as in cell signaling in some tumors . We and others have shown that P15941 expression is activated by progesterone ( P ) , P01375 , and interferon-gamma ( P01579 ) . Here we demonstrate that P15941 expression is down-regulated by overexpression of members of the protein inhibitor of activated signal transducer and activator of transcription ( PIAS ) family , O75925 , Q9Y6X2 , PIASxalpha , PIASxbeta , and Q8N2W9 , in human uterine epithelial cell lines DB09106 and O14777 -1A and in a breast cancer cell line , T47D . Treatments with P , P01375 , and P01579 were unable to overcome the repression by Q8N2W9 . Q8N2W9 repression of basal , P- , and P01375 -stimulated P15941 promoter activity was not dependent on the Q8N2W9 sumoylation domain . In contrast , Q8N2W9 suppression of P01579 -activated P15941 promoter activity was dependent on the Q8N2W9 sumoylation domain . Q8N2W9 and P receptor B were localized to the nucleus upon P treatment , and small interfering RNA knockdown of Q8N2W9 resulted in an increase in P-mediated stimulation of P15941 protein expression . Overexpression of Q8N2W9 did not affect P receptor B binding to the P15941 promoter but surprisingly led to a loss of nuclear receptor corepressor ( NCoR ) , which was recruited to the promoter in response to P. Collectively , these data indicate that Q8N2W9 may be a useful target for down-regulation of P15941 expression in various contexts . The role of pemetrexed in advanced non small-cell lung cancer : special focus on pharmacology and mechanism of action . DB00642 is a newer antifolate drug that has been approved as first-line treatment for patients with advanced non-squamous , non-small cell lung cancer ( NSCLC ) in combination with cisplatin , and as single agent for relapsed or chemotherapy refractory NSCLC after platinum-containing chemotherapy , at a dose of 500 mg/m(2) . DB00642 undergoes intracellular activation by poly-gamma-glutamylation , that is essential for its antiproliferative activity . Polyglutamate derivatives mainly inhibit three key enzymes of intracellular folate metabolism , i.e. thymidylates synthase ( P04818 ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyltransferase ( GARFT ) , with P04818 being the most relevant target . DB00642 undergoes rapid renal elimination as unchanged parent compound , with a terminal half-life of between two to five hours . In later clinical development , the usefulness of supplementation with folic acid and vitamin B(12) became evident , to control pemetrexed-related toxicity . The results from the phase III upfront registration study , a retrospective observational data , and a recent maintenance study of pemetrexed in NSCLC suggest histological subtype to be the most important predictive marker for clinical outcome in patients receiving pemetrexed , DB00642 is active in patients with non-squamous cell NSCLC while no benefit is seen in patients with squamous-cell histology , possibly as a result of different expression of intratumoral P04818 . These are important steps towards individualisation of anticancer treatment in patients with advanced NSCLC . DB06212 , a selective oral vasopressin V2 receptor antagonist , ameliorates podocyte injury in puromycin aminonucleoside nephrotic rats . BACKGROUND : Proteinuria caused by glomerular disease is characterized by podocyte injury . P30518 antagonists are effective in reducing albuminuria , although their actions on glomerular podocytes have not been explored . The objective of this study was to evaluate the effects of tolvaptan , a selective oral V2 receptor antagonist , on podocytes in a puromycin aminonucleoside ( PAN ) -induced nephrosis rat model . METHODS : Rats were allocated to a control , PAN nephrosis , or tolvaptan-treated PAN nephrosis group ( n = 9 per group ) . Urinary protein excretion and serum levels of total protein , albumin , creatinine , and total cholesterol were measured on day 10 . The influence of tolvaptan on podocytes was examined in renal tissues by immunofluorescence and electron microscopy . RESULTS : PAN induced massive proteinuria and serum creatinine elevation on day 10 , both of which were significantly ameliorated by tolvaptan . Immunofluorescence studies of the podocyte-associated proteins nephrin and podocin revealed granular staining patterns in PAN nephrosis rats . In tolvaptan-treated rats , nephrin and podocin expressions retained their normal linear pattern . Electron microscopy showed foot process effacement was ameliorated in tolvaptan-treated rats . CONCLUSIONS : DB06212 is protective against podocyte damage and proteinuria in PAN nephrosis . This study indicates that tolvaptan exerts a renoprotective effect by affecting podocyte morphology and probably function in PAN nephrosis . DB06212 is a promising pharmacological tool in the treatment of renal edema . 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 . The impact of biological agents interfering with receptor/ligand binding in the immune system . We herein discuss the impact of biological agents based on the ability of monoclonal antibodies to target specific molecules . This approach has given to clinical immunologists a spectrum of drugs able to manipulate the immune system . In the first session , we discuss drugs targeting T-cell function by : ( 1 ) targeting P10747 mediated costimulation ( DB01281 and DB06681 ) ; ( 2 ) interfering with interleukin-2 receptor ( DB00074 and DB00111 ) ; ( 3 ) blocking cell adhesion and homing ( DB00092 , DB00095 , DB00108 ) . The second session is dedicated to drugs targeting cytokines or their receptors . The best known and largely experimented case is represented by drugs targeting tumor necrosis factor ( P01375 ) ( DB00065 , Adalilumab , Certolizumab ) or its p75 receptor ( DB00005 ) . However , newer products are now available to target other inflammatory cytokines including P05231 , P10145 , IL-12 , P40933 , Q14116 , IL-23 . These agents have the potential to become powerful tools in the control of several immune-mediated diseases , especially auto-immune and inflammatory ones . They traslate into reality the prediction that antibodies will eventually become " magic bullets which seek their own target " ( P. Ehrich , 1906 ) . Tetrandrine inhibits proinflammatory cytokines , P35228 and P35354 expression in human monocytic cells . Tetrandrine ( TET ) , a bis-benzylisoquinoline alkaloid isolated from the dried root of hang-fang-chi ( Stephania tetrandra S. Moore ) , is traditionally used in China for treating inflammation , hypertension and silicosis . In this study , our aim was to examine the anti-inflammatory mechanism of TET through measuring the inducible nitric oxide synthase ( P35228 ) , cyclooxygenase-1 , and -2 ( P23219 and P35354 ) expression , cytokines ( P01375 , P05112 and P10145 ) formation , nitric oxide ( NO ) release and prostaglandin E2 ( DB00917 ) generation in lipopolysaccharide ( LPS ) -induced human monocytic ( THP-1 ) cells . Results showed that TET remarkably suppressed the LPS ( 1 microg/ml ) induction of NO release and DB00917 generation . It also significantly attenuated the LPS-induced transcription of proinflammatory cytokines ( P01375 , P05112 and P10145 ) in a dose-dependent manner . Furthermore , TET at 100 microM significantly blocked the LPS induction of P35228 and P35354 expression , but not the P23219 . Taken together , these results suggest that TET exerts anti-inflammatory effects probably through the suppression of P35354 and P35228 expression . [ Cell cycle analysis of endometrial cancer cells in vitro treated with growth factor and steroid hormone ] . The aim of this study was to overtake the mechanism of the control system in endometrial cancer cell line in vitro . Ishikawa cell ( IK cell ) and O14777 -1 cell ( O14777 cell ) derived from endometrial cancers were cultured with serum free medium ( SFM-101 ) . IK cell possessed P03372 ( ER ) , P06401 ( PR ) , Epidermal growth factor ( P01133 ) and its receptor ( P00533 ) . O14777 cell had PR , P01133 , and P00533 , however O14777 cell did not keep ER . P01133 stimulated the growth of IK cell , but the growth of O14777 cell was not stimulated by P01133 . S phase cells were increased by P01133 in IK cell , but were not increased by P01133 in O14777 cell . The growth of IK cell was stimulated significantly by P01133 and Estradiol-17 beta ( E2 ) + P01133 than control . However , E2+ P01133 did not stimulate the growth of IK cell than P01133 significantly . DB01406 ( D ) and D+ P01133 inhibited the growth of IK cell significantly than control . S phase cells were decreased by the treatment of D and D+ P01133 . From our results , P01133 stimulated the growth of ER positive endometrial cancer cell , but P01133 did not stimulate ER negative endometrial cancer cell . E2+ P01133 and P01133 stimulated the growth of IK cell as a same . However , D inhibited the growth of IK cell that was stimulated by P01133 . Effects of pemetrexed , gefitinib , and their combination on human colorectal cancer cells . PURPOSE : The study investigated the effects of pemetrexed , gefitinib , and their combination on human colorectal cancer cells . METHODS : Six human colorectal cancer cells were exposed to pemetrexed , gefitinib , and their combination . Antitumor effects were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay . P04818 ( TS ) mRNA expression and P00533 mutation were studied by real-time RT-PCR and DNA sequence determination . Pharmacological interaction was studied using the combination index method . Cell cycle distribution and apoptosis were determined by flow cytometry . Activity assay was performed to assess the effects of drugs on TS activity , and Western blot was performed to assess the protein expression of pEGFR , pAKT , and pERK 1/2 . RESULTS : Six colorectal cancer cells are all sensitive to pemetrexed , and TS gene expression of cells was negatively related to pemetrexed sensitivity . The cytotoxic synergism was observed in concurrent pemetrexed combined with gefitinib and sequential pemetrexed followed by gefitinib . The combination of pemetrexed and gefitinib modulated cell cycle and induced apoptosis . DB00642 combined with gefitinib decreased TS mRNA expression and in situ activity . DB00642 induced an P00533 -mediated activation of the phosphatidylinositol 3-kinase/AKT and P29323 pathway , which was inhibited by gefitinib . CONCLUSIONS : DB00642 is a promising agent , and pemetrexed combined with gefitinib has a significantly synergistic effect on colorectal cancer cells , which seems to present a strategy of pemetrexed combined with P00533 -TKIs in colorectal cancer treatment . Rosiglitazone regulates P05231 -stimulated lipolysis in porcine adipocytes . Interleukin ( IL ) -6 , a proinflammatory cytokine , stimulates adipocyte lipolysis and induces insulin resistance in obese and diabetic subjects . However , the effects of the anti-diabetic drug rosiglitazone on P05231 -stimulated lipolysis and the underlying molecular mechanism are largely unknown . In this study , we demonstrated that rosiglitazone suppressed P05231 -stimulated lipolysis in differentiated porcine adipocytes by inactivation of extracellular signal-related kinase ( P29323 ) . Meanwhile , rosiglitazone enhanced the lipolysis response of adipocytes to isoprenaline . In addition , rosiglitazone significantly reversed P05231 -induced down-regulation of several genes such as perilipin A , peroxisome proliferators activated receptor gamma ( Q07869 & gamma ; ) , and fatty acid synthetase , as well as the up-regulation of P05231 mRNA . However , mRNA expression of Q07869 & gamma ; coactivator-1 alpha ( DB01053 -1 & alpha ; ) was enhanced by rosiglitazone in P05231 -stimulated adipocytes . These results indicate that rosiglitazone suppresses P05231 -stimulated lipolysis in porcine adipocytes through multiple molecular mechanisms . Toward a chronoimmunomodulation by cefodizime in multiple myeloma and chronic uremia . Nine healthy subjects , 19 patients with multiple myeloma ( MM ) and 21 patients with chronic uremia were given cefodizime ( 2 g i.v. ) at 2 different timepoints either in the morning or in the evening for 5 to 7 consecutive days . The following immunological parameters were comparatively evaluated before and after cefodizime administration : rosette-forming cells ( P41440 % ) , T lymphocyte subpopulations , monocyte chemotaxis index ( D6RGH6 ) and granulocyte chemotaxis index ( GCI ) . Independently of the time of drug administration , a circadian rhythm was clearly detected ( 90 % CI ) as regards P41440 % , CD3 , P01730 , CD8 , P01730 /CD8 before and P41440 % , CD3 , P01730 , CD8 , GCI after therapy . In addition , in patients treated at 0800 cefodizime increased the MESOR of the D6RGH6 and , to a lesser extent , of the GCI . The chronoimmunomodulatory effects of cefodizime in patients with MM and chronic uremia are discussed . Prevalence of insomnia and associated factors in South Korea . INTRODUCTION : In Western countries , insomnia is associated with daytime impaired functioning , as well as physical and psychiatric illnesses . However , little information exists on insomnia in Asian countries . This study investigates the prevalence and correlates of insomnia in the general population of South Korea . METHODS : A representative sample of the South Korean general population composed of 3719 noninstitutionalized individuals aged 15 years or older were interviewed by telephone using the Sleep-EVAL system . The participation rate was 91.4 % . The interviews covered sleep habits , sleep symptomatology , physical and psychiatric illnesses . DSM-IV sleep and psychiatric disorder diagnoses were also assessed . RESULTS : Insomnia symptoms occurring at least three nights per week were reported by 17.0 % of the sample ; difficulty initiating sleep ( Q8IX12 ) was mentioned by 4.0 % of the sample , difficulty maintaining sleep ( DMS ) by 11.5 % , early morning awakenings ( P15941 ) by 1.8 % , and nonrestorative sleep ( NRS ) by 4.7 % of the sample . DSM-IV insomnia disorder diagnoses were found in 5 % of the sample . Over 50 % of subjects with insomnia symptoms reported important daytime consequences and another 20 % reported mild or moderate consequences . However , the proportion of insomnia subjects seeking medical help for their sleep problems was very low ( 6.8 % ) . CONCLUSIONS : As in Western countries , insomnia is widespread in South Korea , affecting nearly one in five individuals . Many of them would benefit from medical help ; however , few insomnia subjects are consulting for this problem . An educational effort is needed for both the general population and the physicians . Effects of the total saponins from Rosa laevigata Michx fruit against acetaminophen-induced liver damage in mice via induction of autophagy and suppression of inflammation and apoptosis . The effect of the total saponins from Rosa laevigata Michx fruit ( RLTS ) against acetaminophen ( DB00316 ) -induced liver damage in mice was evaluated in the present paper . The results showed that RLTS markedly improved the levels of liver SOD , CAT , DB00143 , DB00143 -Px , MDA , NO and P35228 , and the activities of serum ALT and Q9NRA2 caused by DB00316 . Further research confirmed that RLTS prevented fragmentation of DNA and mitochondrial ultrastructural alterations based on TdT-mediated dUTP nick end labeling ( TUNEL ) and transmission electron microscopy ( TEM ) assays . In addition , RLTS decreased the gene or protein expressions of cytochrome P450 ( P05181 ) , pro-inflammatory mediators ( IL-1β , P05112 , P05231 , P01375 -α , P35228 , Bax , HMGB-1 and P35354 ) , pro-inflammatory transcription factors ( NF-κB and AP-1 ) , pro-apoptotic proteins ( cytochrome C , p53 , caspase-3 , caspase-9 , p-JNK , p-p38 and p- P29323 ) , and increased the protein expressions of Bcl-2 and Bcl-xL . Moreover , the gene expression of P22301 , and the proteins including LC3 , Q14457 and Atg5 induced by DB00316 were even more augmented by the extract . These results demonstrate that RLTS has hepatoprotective effects through antioxidative action , induction of autophagy , and suppression of inflammation and apoptosis , and could be developed as a potential candidate to treat DB00316 -induced liver damage in the future . Characterization of the pattern of the nongenomic signaling pathway through which TCDD-induces early inflammatory responses in U937 human macrophages . 2,3,7,8-Tetrachlorodibenzo(p)dioxin ( TCDD ) has been known to induce inflammatory signaling in a number of cell types and tissues . We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 ( P47712 ) within 30min as judged by the increase in the serine 505 phosphorylated form of P47712 protein and the increased cellular release of free arachidonic acid . This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker , P35354 mRNA expression within 1h , and by 3h , several other markers become up-regulated . These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+) , and activation of P47712 and P35354 . A comparative study among three different human cell lines showed that activation of P35354 within 1h of action of TCDD is a common feature exhibited by all cell lines . On the other hand , the U937 macrophage line appears to be unique among them with respect to its ability to activate P01375 and P10145 mRNA expressions , and not requiring Src kinase in propagating the initial signaling of P47712 . Based on the rapidity of activation of P47712 and P35354 , which occurs within 1h of cell exposure to TCDD , when no change in mRNA expression of P04798 has been observed , it is apparent that this unique action of TCDD is carried out through a distinct " nongenomic " pathway which , is clearly discernable from the classical , " genomic " action pathway of the P35869 by not requiring the participation of P27540 . Release of cytokines by blood monocytes during strenuous exercise . During strenuous exercise in endurance athletes , monocytes are activated and there is an acute inflammation and hypoxemia possibly due to lesional pulmonary edema . P05231 and P01375 released by monocytes may be implicated in the acute phase of lesional pulmonary edema . A study was carried out to determine whether P01375 and P05231 are released during strenuous exercise , and , if adrenalin released during exercise alters their generation . Ten young and six master athletes underwent an incremental exercise test . Arterial blood was drawn at rest , at the end of the exercise , and 20 minutes afterwards . Monocytes were isolated and incubated for 18 hours in the presence or absence of adrenalin . Il-6 and P01375 were measured in monocyte supernatants . The spontaneous release of P05231 or P01375 was increased in young athletes when compared to older subjects . The spontaneous release of P01375 was increased , but not significantly , by exercise and there was no correlation between the release of P05231 and P01375 and lung function measured during hypoxemia . DB00668 inhibited the release of P05231 or P01375 . Correlations were observed between the in vitro release of P05231 or P01375 and age , VO2max , maximal ventilation and maximal power output of the subjects . DB00642 : its promise in treating non-small-cell lung cancer . The use of chemotherapy in the treatment of early and advanced non-small-cell lung cancer ( NSCLC ) has increased during the past decade . One of the main reasons for the increased acceptance of chemotherapy is the development of several new cytotoxic agents with a unique mechanism(s) of action and high single-agent activity , combined with a favorable toxicity profile . DB00642 ( Alimta ) is a novel antifolate that inhibits several enzymes involved in DNA synthesis ( thymidylate synthase [ TS ] , dihydrofolate reductase [ P00374 ] , and glycinamide ribonucleotideformyltransferase [ GARFT ] ) . DB00642 's toxicity is markedly reduced by folic acid and vitamin B12 supplementation . The compound has been studied extensively in various tumor types , including NSCLC . In NSCLC , pemetrexed at 500 mg/m2 , every 3 weeks , given i.v. over 10 minutes , has shown promising activity , and can safely be administrated with vitamin supplementation . After registration , single-agent pemetrexed will certainly add to the chemotherapeutic options available for pretreated patients and will most likely change significantly chemotherapy prescriptions in second-line chemotherapy . In first-line chemotherapy , the role of platinum-based and -free combination doublet chemotherapy with pemetrexed still needs to be defined . Phase II data indicate high efficacy combined with favorable toxicity for pemetrexed in combination with cisplatin , carboplatin ( DB00958 ) , oxaliplatin ( Eloxatin ) , gemcitabine ( Gemzar ) , and vinorelbine ( Navelbine ) . This review summarizes the clinical experience obtained thus far during the early clinical development of pemetrexed in NSCLC . Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum . Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease . Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this Q07869 agonist . Anion-exchange solid-phase extraction , in combination with reverse-phase high-performance liquid chromatography ( HPLC ) , rapidly and accurately determines steady-state fenofibric acid serum concentrations . Chromatographic separation under isocratic conditions , with use of ultraviolet detection at 285 nm , provides clean baseline and sharp peaks for clofibric acid , 1-napthyl acetic acid ( internal standards ) , and fenofibric acid . Commonly prescribed and over-the-counter nonsteroidal anti-inflammatory drugs ( NSAIDs ) were screened for assay interference , and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens . DB01039 analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard . Accuracies ranged from 98.65 % to 102.4 % , whereas the within- and between-day precisions ranged from 1.0 % to 2.2 % and 2.0 % to 6.2 % , respectively . NSAIDs had minimal interference with the assay , which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects , many taking a variety of coadministered medications . Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications . This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia . 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 . Translational research with pemetrexed in breast cancer . DB00642 ( Alimta ) is a novel folate antimetabolite that primarily inhibits the enzymes thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , and glycinamide ribonucleotide formyl transferase ( GARFT ) , all of which are involved in pyrimidine and purine synthesis . In a phase II trial of patients with DB00279 /4 , N0-2 breast cancer , expression of thymidylate synthase ( TS ) , dihydrofolate reductase ( P00374 ) , glycinamide ribonucleotide formyltransferase ( GARFT ) , p53 , and c-erb-B2 ( at the mRNA or protein level ) was examined in tumor biopsy specimens before and 24 hours after the first dose of pemetrexed and after three cycles of single-agent treatment to establish correlations of biomarker levels and changes with clinical outcome and toxicity . Although final data are not available , initial indications are that clinical response may correlate with decreased or low TS expression . The results obtained from clinical data are supported by laboratory results in three cell lines ( MDA-231 , MCF-7 , and ZR-75 ) . These results suggest that in vitro transcript profiling to identify which genes are important predictors of successful cytotoxic chemotherapy , followed by a focused clinical trial to confirm the in vitro results , may be the best approach for translational research . Differential involvement of Galpha16 in CC chemokine-induced stimulation of phospholipase Cbeta , P29323 , and chemotaxis . Chemokines are known to regulate the chemotaxis of leukocytes and play an important role in immunological processes . Chemokine receptors are widely distributed in hematopoietic cells and are often co-localized with the hematopoietic-specific G(16) and its close relative , G(14) . Yet , many chemokine receptors utilize pertussis toxin-sensitive G(i) proteins for signaling . Given that both G(16) and G(14) are capable of linking G(i)-coupled receptors to the stimulation of phospholipase Cbeta , we examined the capacity of six CC chemokine receptors ( P32246 , CCR2a , CCR2b , P51677 , P51681 and P32248 ) to interact with G(14) and G(16) in a heterologous expression system . Among the CC chemokine receptors tested , P32246 , CCR2b , and P51677 were capable of mediating chemokine-induced stimulation of phospholipase Cbeta via either G(14) or G(16) . The G(14)/G(16)-mediated responses exhibited CC chemokine dose-dependency and were resistant to pertussis toxin ( PTX ) treatment . In contrast , CCR2a , P51681 and P32248 were unable to interact with G(14) and G(16) . Under identical experimental conditions , all six CC chemokine receptors were fully capable of inhibiting adenylyl cyclase via G(i) as well as stimulating phospholipase Cbeta via 16z44 , a G(16/z) chimera that possesses increased promiscuity toward G(i)-coupled receptors . Moreover , P32246 -mediated P27361 /2 phosphorylation was largely PTX-insensitive in THP-1 monocytic cells that endogenously express Galpha(16) . In addition , P32246 agonist was less efficacious in mediating chemotaxis of THP-1 cells following the knockdown of Galpha(16) by overexpressing siRNA , indicating the participation of Galpha(16) in P32246 -induced cell migration . These results show that different CC chemokine receptors can discriminate against G(14) and G(16) for signal transduction .